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Sample records for anticuerpos igm al

  1. Diagnóstico diferencial en la encefalitis por anticuerpos contra el receptor NMDA

    PubMed Central

    González-Valcárcel, J.; Rosenfeld, M.R.; Dalmau, J.

    2011-01-01

    Resumen Introducción La encefalitis por anticuerpos contra el receptor de NMDA (NMDAR) suele desarrollarse como un síndrome característico de evolución multifásica y diagnóstico diferencial amplio. Pacientes Presentamos a 2 pacientes diagnosticadas de encefalitis por anticuerpos NMDAR con un cuadro clínico típico, pero que inicialmente señaló otras etiologías. Discusión La afectación frecuente de pacientes jóvenes con manifestaciones psiquiátricas prominentes indica frecuentemente otras consideraciones diagnósticas; las más frecuentes son las encefalitis virales, los procesos psiquiátricos y el síndrome neuroléptico maligno. Varios síndromes previamente definidos de manera parcial o descriptiva en adultos y pacientes pediátricos probablemente eran casos de encefalitis anti-NMDAR. Conclusiones La encefalitis anti-NMDAR debe considerarse en pacientes jóvenes con manifestaciones psiquiátricas subagudas, movimientos anormales y alteraciones autonómicas. La caracterización clínica e inmunológica de esta enfermedad ha llevado a la identificación de nuevos anticuerpos que afectan a procesos de memoria, aprendizaje, conducta y psicosis. PMID:20964986

  2. Anticuerpos derivados de pacientes se dirigen a células tumorales

    Cancer.gov

    Artículo del blog Temas y relatos de un anticuerpo derivado de pacientes que destruye células tumorales en estirpes celulares de varios tipos de cáncer e hizo lento el crecimiento en modelos murinos de cáncer de cerebro y pulmón sin signo de efectos.

  3. IgM nephropathy; time to act.

    PubMed

    Mubarak, Muhammed

    2014-01-01

    Implication for health policy/practice/research/medical education: Much has been published on the epidemiology and clinicopathological characteristics of IgM nephropathy, but there is little information on the etiology,pathogenesis and specific therapy of the disease. Controversy still shrouds the definition and nosologic status of the disease. Well-coordinated and concerted international efforts and collaboration between researchers in the developing and developed countries are needed to make further progress on the above aspects of the disease. PMID:24644539

  4. False Positive Lyme Disease IgM Immunoblots in Children.

    PubMed

    Lantos, Paul M; Lipsett, Susan C; Nigrovic, Lise E

    2016-07-01

    In our cross-sectional sample of 7289 serologic tests for Lyme disease, we identified 167 instances of a positive IgM immunoblot but a negative IgG immunoblot test result. Considering that only 71% (95% CI 64%-78%) of patients had Lyme disease, a positive IgM immunoblot alone should be interpreted with caution to avoid over-diagnosis of Lyme disease.

  5. Mass loss from galaxies: feeding the IGM, recycling in the IGM

    NASA Astrophysics Data System (ADS)

    Duc, P.-A.; Braine, J.; Lisenfeld, U.; Amram, P.; Brinks, E.

    2002-07-01

    As a result of internal processes or environmental effects like ram-pressure stripping or collisions, galaxies lose a significant part of their stellar and gaseous content. Whereas the impact of such stripping on galaxy evolution has been well studied, much less attention has been given to the fate of the expelled material in the intergalactic or intra cluster medium (IGM/ICM). Observational evidence exists showing that a fraction of the injected matter is actually recycled to form a new generation of galaxies, such as the Tidal Dwarf Galaxies discovered near numerous interacting systems. Using a set of multiwavelength data, we are now able to roughly analyze the processes pertaining to their formation: from an instability in the HI clouds, through the formation of molecular gas, and to the onset of star formation.

  6. Evaluation of the IMMULITE® 2000 CMV IgM assay

    PubMed Central

    2012-01-01

    Background Diagnosis of cytomegalovirus (CMV) infection is challenging because of the high rate of asymptomatic infection and the low specificity of associated symptoms and signs. As a result, laboratory testing is an essential aid in making an accurate diagnosis. The presence of CMV IgM is indicative of primary CMV infection. In pregnancy, diagnosis of primary infection is important because primary maternal infection increases fetal infection risk substantially. Fetal infection can result in serious sequelae ranging from neurological deficits to death. Diagnosis among the immunocompromised is also critical for the timely initiation of therapy that can reduce morbidity and mortality risk. Methods The IMMULITE® 2000 CMV IgM assay qualitatively detects CMV IgM antibodies in human serum or plasma to aid in the diagnosis of current or recent CMV infection. To determine expected values in apparently healthy subjects, 136 samples were tested. Reproducibility, normal range, and method comparison studies were also performed to evaluate the assay's performance. The assay's reproducibility was evaluated across three sites. Seven hundred and eighteen (n = 718) individual patient serum samples, which included samples from CMV IgM-positive (n = 109, determined by the Abbott IMx CMV or the Diamedix CMV IgM assays), pregnant (n = 210), HIV-positive (n = 30), immunosuppressed (n = 102), and transplant patients (n = 17) and from patients with potentially cross-reacting conditions (n = 136) were evaluated in the method comparison study. The positive, negative, and overall agreement between the IMMULITE 2000 CMV IgM assay and the VIDAS CMV IgM assay (predicate assay) were determined. Results The assay demonstrated excellent reproducibility with a total CV of less than 10%. The positive, negative, and overall agreement between the IMMULITE 2000 assay and the VIDAS assay were > 95% for the method comparison samples. Among potentially cross-reactive samples, the overall agreement

  7. A Reassessment of IgM Memory Subsets in Humans.

    PubMed

    Bagnara, Davide; Squillario, Margherita; Kipling, David; Mora, Thierry; Walczak, Aleksandra M; Da Silva, Lucie; Weller, Sandra; Dunn-Walters, Deborah K; Weill, Jean-Claude; Reynaud, Claude-Agnès

    2015-10-15

    From paired blood and spleen samples from three adult donors, we performed high-throughput VH sequencing of human B cell subsets defined by IgD and CD27 expression: IgD(+)CD27(+) ("marginal zone [MZ]"), IgD(-)CD27(+) ("memory," including IgM ["IgM-only"], IgG and IgA) and IgD(-)CD27(-) cells ("double-negative," including IgM, IgG, and IgA). A total of 91,294 unique sequences clustered in 42,670 clones, revealing major clonal expansions in each of these subsets. Among these clones, we further analyzed those shared sequences from different subsets or tissues for VH gene mutation, H-CDR3-length, and VH/JH usage, comparing these different characteristics with all sequences from their subset of origin for which these parameters constitute a distinct signature. The IgM-only repertoire profile differed notably from that of MZ B cells by a higher mutation frequency and lower VH4 and higher JH6 gene usage. Strikingly, IgM sequences from clones shared between the MZ and the memory IgG/IgA compartments showed a mutation and repertoire profile of IgM-only and not of MZ B cells. Similarly, all IgM clonal relationships (among MZ, IgM-only, and double-negative compartments) involved sequences with the characteristics of IgM-only B cells. Finally, clonal relationships between tissues suggested distinct recirculation characteristics between MZ and switched B cells. The "IgM-only" subset (including cells with its repertoire signature but higher IgD or lower CD27 expression levels) thus appear as the only subset showing precursor-product relationships with CD27(+) switched memory B cells, indicating that they represent germinal center-derived IgM memory B cells and that IgM memory and MZ B cells constitute two distinct entities.

  8. Hyper IgM Syndrome with low IgM and thrombocytosis: an unusual case of immunodeficiency.

    PubMed

    Yousef, Ejaz; Arshad Alvi, M

    2016-09-01

    We report a 5 years old male child with low serum IgG, IgA and IgM levels, who presented with recurrent perianal and oral ulcers, intermittent fever, and protracted diarrhea. Despite the lack of typical respiratory symptoms, low serum IgM level and persistent thrombocytosis, an X-linked hyper-IgM syndrome (X-HIGM) was considered. Laboratory investigations revealed a diagnosis of hyper-IgM syndrome caused by CD40L deficiency. PMID:27608476

  9. Encefalitis por anticuerpos contra el receptor de NMDA: experiencia con seis pacientes pediátricos. Potencial eficacia del metotrexato

    PubMed Central

    Bravo-Oro, Antonio; Abud-Mendoza, Carlos; Quezada-Corona, Arturo; Dalmau, Josep; Campos-Guevara, Verónica

    2016-01-01

    Introducción La encefalitis por anticuerpos contra el receptor de N-metil-D-aspartato (NMDA) es una entidad cada vez más diagnosticada en edad pediátrica. A diferencia de los adultos, en muchos casos no se asocia a tumores y las manifestaciones iniciales en niños más frecuentes son crisis convulsivas y trastornos del movimiento, mientras que en los adultos predominan las alteraciones psiquiátricas. Casos clínicos Presentamos seis casos pediátricos confirmados con anticuerpos contra la subunidad NR1 del receptor de NMDA en suero y líquido cefalorraquídeo. Cinco de los casos comenzaron con crisis convulsivas como manifestación clínica inicial antes de desarrollar el cuadro clásico de esta entidad. En todos los casos se utilizaron esteroides como primera línea de tratamiento, con los que sólo se observó control de las manifestaciones en uno, por lo que el resto de los pacientes requirió inmunomoduladores de segunda línea. Todos los pacientes recibieron metotrexato como tratamiento inmunomodulador para evitar recaídas y la evolución fue a la mejoría en todos ellos. Conclusiones En nuestra serie de pacientes con encefalitis por anticuerpos contra el receptor de NMDA, ninguno se asoció a tumores. Todos los casos recibieron metotrexato por lo menos durante un año, no observamos eventos adversos clínicos ni por laboratorio, ni hubo secuelas neurológicas ni recaídas durante el tratamiento. Aunque es una serie pequeña y es deseable incrementar el número y tiempo de evolución, consideramos el metotrexato una excelente alternativa como tratamiento inmunomodulador para esta patología. PMID:24150952

  10. A Reassessment of IgM Memory Subsets in Humans.

    PubMed

    Bagnara, Davide; Squillario, Margherita; Kipling, David; Mora, Thierry; Walczak, Aleksandra M; Da Silva, Lucie; Weller, Sandra; Dunn-Walters, Deborah K; Weill, Jean-Claude; Reynaud, Claude-Agnès

    2015-10-15

    From paired blood and spleen samples from three adult donors, we performed high-throughput VH sequencing of human B cell subsets defined by IgD and CD27 expression: IgD(+)CD27(+) ("marginal zone [MZ]"), IgD(-)CD27(+) ("memory," including IgM ["IgM-only"], IgG and IgA) and IgD(-)CD27(-) cells ("double-negative," including IgM, IgG, and IgA). A total of 91,294 unique sequences clustered in 42,670 clones, revealing major clonal expansions in each of these subsets. Among these clones, we further analyzed those shared sequences from different subsets or tissues for VH gene mutation, H-CDR3-length, and VH/JH usage, comparing these different characteristics with all sequences from their subset of origin for which these parameters constitute a distinct signature. The IgM-only repertoire profile differed notably from that of MZ B cells by a higher mutation frequency and lower VH4 and higher JH6 gene usage. Strikingly, IgM sequences from clones shared between the MZ and the memory IgG/IgA compartments showed a mutation and repertoire profile of IgM-only and not of MZ B cells. Similarly, all IgM clonal relationships (among MZ, IgM-only, and double-negative compartments) involved sequences with the characteristics of IgM-only B cells. Finally, clonal relationships between tissues suggested distinct recirculation characteristics between MZ and switched B cells. The "IgM-only" subset (including cells with its repertoire signature but higher IgD or lower CD27 expression levels) thus appear as the only subset showing precursor-product relationships with CD27(+) switched memory B cells, indicating that they represent germinal center-derived IgM memory B cells and that IgM memory and MZ B cells constitute two distinct entities. PMID:26355154

  11. Enhanced in vitro IgM rheumatoid factor synthesis in seropositive rheumatoid arthritis families.

    PubMed

    Alarcón, G S; Barger, B O; Go, R C; Acton, R T; Schrohenloher, R E; Koopman, W J

    1987-01-01

    We studied the patterns of in vitro IgM RF production by peripheral blood mononuclear leukocytes (MNL) in first- and second-degree relatives and non-blood relatives of 14 RA patients. In all these included 25 individuals with RA (14 probands, 11 other affected members), 88 non-affected first- and second-degree relatives and 15 non-affected non-blood relatives. In vitro production of IgM RF and IgM by MNL from RA patients was similar to what we and others have previously described. Spontaneous production of IgM RF was demonstrated by MNL from some RA patients and the ratio of IgM RF to total IgM was higher for patients than controls. The non-blood relatives failed to demonstrate spontaneous in vitro IgM RF production. In contrast, MNL from approximately 16% of non-affected blood relatives spontaneously produced IgM RF. Spontaneous IgM RF production in this group did not correlate with HLA-DR3 or HLA-DR4 and was found in Caucasoids. American blacks, simplex (one affected individual), and multiplex (more than one affected individual) families. Linkage analysis using either a recessive or dominant model of inheritance failed to detect linkage among affected and non-affected family members.

  12. The contribution of naturally occurring IgM antibodies, IgM cross-reactivity and complement dependency in murine humoral responses to pneumococcal capsular polysaccharides.

    PubMed

    Jones, Hannah E; Taylor, Philip R; McGreal, Eamon; Zamze, Susanne; Wong, Simon Y C

    2009-09-25

    Immunogenicity of 12 capsular polysaccharides (CPS) from Streptococcus pneumoniae did not correlate with pre-existing levels of natural IgM anti-CPS antibodies in mice. Immunization of mice with individual CPS, with the exception of type 14 (the only neutral CPS tested), increased serum IgM that also bound other CPS serotypes independent of structural similarity or commonly known contaminants. Surprisingly only IgM response to type 4 (which has a small immunodominant epitope) was dependent on either complement C3 or complement receptors CD35/CD21. IgG anti-CPS responses were infrequently induced, but critically dependent on complement. Our results have clarified the role of complement in the induction of IgM and IgG anti-CPS antibody responses in mice and have implications for CPS vaccine development.

  13. The contribution of naturally occurring IgM antibodies, IgM cross-reactivity and complement dependency in murine humoral responses to pneumococcal capsular polysaccharides.

    PubMed

    Jones, Hannah E; Taylor, Philip R; McGreal, Eamon; Zamze, Susanne; Wong, Simon Y C

    2009-09-25

    Immunogenicity of 12 capsular polysaccharides (CPS) from Streptococcus pneumoniae did not correlate with pre-existing levels of natural IgM anti-CPS antibodies in mice. Immunization of mice with individual CPS, with the exception of type 14 (the only neutral CPS tested), increased serum IgM that also bound other CPS serotypes independent of structural similarity or commonly known contaminants. Surprisingly only IgM response to type 4 (which has a small immunodominant epitope) was dependent on either complement C3 or complement receptors CD35/CD21. IgG anti-CPS responses were infrequently induced, but critically dependent on complement. Our results have clarified the role of complement in the induction of IgM and IgG anti-CPS antibody responses in mice and have implications for CPS vaccine development. PMID:19660585

  14. Investigation Of The Diffuse IGM By Cross-Correlation Studies

    SciTech Connect

    Farnsworth, Damon; Brown, Shea; Rudnick, Lawrence

    2009-12-18

    We present results from the first cross-correlation search for the synchrotron component of the diffuse intergalactic medium (IGM) in filamentary large scale structure (LSS). We used the low resolution (36') Bonn survey at 21cm, with the infrared 2MASS catalog as a tracer of the LSS. Synchrotron emission likely results from LSS formation shocks and feedback from AGN and galactic winds [2]. We determined 3{sigma} upper limits to the diffuse emission in units of flux per galaxy; these correspond to filament equipartition magnetic fields as low as 0.2 {mu}G. The detection threshold for the average (peak) filament brightness is 1 (7) mK for 0.03

  15. Competitive binding of pentraxins and IgM to newly exposed epitopes on late apoptotic cells.

    PubMed

    Ciurana, Caroline L F; Hack, C Erik

    2006-01-01

    A random distribution of phospholipids among the inner and outer leaflet of the cell membrane occurs during apoptosis and is known as membrane flip-flop. Flip-flopped cells have binding sites for various plasma proteins, such as IgM and the pentraxins C-reactive protein (CRP) and serum amyloid P component (SAP). In this study, we investigated whether pentraxins and IgM antibodies recognize the same binding sites on apoptotic cells, and whether phospholipids constitute these binding sites. Except for SAP which also bound to early apoptotic cells, pentraxins and IgM preferentially bound to late apoptotic cells. Competition experiments with different phosphatemonoesters revealed that CRP and SAP as well as part of the IgM bound to the phospholipids head groups, SAP mainly to phosphorylethanolamine, CRP to phosphorylcholine and phosphorylethanolamine and to a lesser extent to phosphorylserine, and IgM to phosphorylcholine and phosphorylserine. These results were confirmed in experiments in which proteins were adsorbed from plasma with artificial phospholipids particles. IgM and the pentraxins variably competed for the same binding sites on late apoptotic cells, SAP having the highest and CRP the lowest apparent affinity. We conclude that CRP, SAP, and part of the IgM bind to the phospholipid head groups exposed on apoptotic cells. This shared specificity as well as their shared capability to activate complement, suggest that IgM and the pentraxins CRP and SAP exert similar functions in the removal of apoptotic cells.

  16. The long elusive IgM Fc receptor, FcμR.

    PubMed

    Kubagawa, Hiromi; Oka, Satoshi; Kubagawa, Yoshiki; Torii, Ikuko; Takayama, Eiji; Kang, Dong-Won; Jones, Dewitt; Nishida, Naonori; Miyawaki, Toshio; Bertoli, Luigi F; Sanders, Sheila K; Honjo, Kazuhito

    2014-07-01

    IgM exists as both a monomer on the surface of B cells and a pentamer secreted by plasma cells. Both pre-immune "natural" and antigen-induced "immune" IgM antibodies are important for protective immunity and for immune regulation of autoimmune processes by recognizing pathogens and self-antigens. Effector proteins interacting with the Fc portion of IgM, such as complement and complement receptors, have thus far been proposed but fail to fully account for the IgM-mediated protection and regulation. A major reason for this deficit in our understanding of IgM function seems to be lack of data on a long elusive Fc receptor for IgM (FcμR). We have recently identified a bona fide FcμR in both humans and mice. In this article we briefly review what we have learned so far about FcμR.

  17. False-positive results in immunoglobulin M (IgM) toxoplasma antibody tests and importance of confirmatory testing: the Platelia Toxo IgM test.

    PubMed Central

    Liesenfeld, O; Press, C; Montoya, J G; Gill, R; Isaac-Renton, J L; Hedman, K; Remington, J S

    1997-01-01

    Although tests for detection of immunoglobulin M (IgM) toxoplasma antibodies have been reported to have a high degree of accuracy, it is well recognized by investigators in the United States and Europe that false-positive results may occur with many of these tests, at times to an alarming degree. Unfortunately, this information is not well documented in the literature. Studies on various toxoplasma IgM test kits are frequently flawed. The investigators often use reference tests which have not previously been carefully evaluated as well as sera that were not appropriate to answer the question of how often false-positive results might occur. We recently had the unique opportunity to evaluate the accuracy of the Platelia Toxo IgM test in 575 serum samples obtained during an outbreak of toxoplasmosis which occurred in 1995 in the Capital Regional District of British Columbia, Canada. When compared with results obtained in a reference IgM enzyme-linked immunosorbent assay (ELISA), the Platelia Toxo IgM test had a sensitivity of 99.4%, specificity of 49.2%, positive predictive value of 51.9%, negative predictive value of 99.3%, and an overall agreement of 67.0%. In an attempt to resolve discrepancies between these two tests, a serological profile (Sabin-Feldman dye test, IgA and IgE antibody tests, differential agglutination [AC/HS] test, and IgG avidity method) was performed. Of 153 serum samples that were positive in the Platelia Toxo IgM test and negative in the IgM ELISA, 71 (46.4%) were negative in the Sabin-Feldman dye test. Of the serum samples that were positive in the dye test, 77 (93.9%) had a serological profile most compatible with an infection acquired in the distant past. These results reveal high numbers of false-positive results in the Platelia Toxo IgM test and highlight the importance of appropriate evaluation of commercial tests that are currently being marked. Our results also emphasize the importance of confirmatory testing to determine whether the

  18. The quest to detect X-ray IGM

    NASA Astrophysics Data System (ADS)

    Rasmussen, A.; Paerels, F.; Kahn, S. M.; den Herder, J.-W.; de Vries, C.

    2004-08-01

    Several efforts have been made since the availability of Chandra and XMM/Newton observatories to find evidence for a putative diffuse IGM at X-ray temperatures that trace the total mass distribution in the local universe. While careful analysis of X-ray emission from the outer regions of clusters of galaxies sometimes reveal presence of marginally more soft X-ray emission than expected (AKA the soft excess phenomenon), spectroscopic measures to detect line absorption by intervening systems toward bright, backlight AGNs have led only to very tentative detections that beg better statistics. Using megaseconds of archival XMM/Newton RGS data, combined with high quality spectroscopic data that probe structure of the local ISM, we characterize the extragalactic component of the absorption spectrum toward three lines of sight previously reported (3C273, PKS2155-304 and Mrk 421). We will also briefly describe preliminary design parameters for YADEX: a wide field, X-ray redshift survey mission that may stand a fighting chance to map the mass distribution structure in the local universe.

  19. Detecting the Warm-Hot IGM via Ultraviolet Absorbers

    NASA Astrophysics Data System (ADS)

    Danforth, Charles

    2010-03-01

    Detecting the Warm-Hot Intergalactic Medium (WHIM, T=105-107 K) via X-ray absorption in highly-ionized metal ions is a major goal in X-ray astronomy today. Species such as OVII and OVIII are predicted to be present in large quantities in the IGM, tracing metal-enriched diffuse gas at T>106 K. This reservoir of WHIM may make up a substantial fraction of the "missing" baryons in the low-redshift universe, however detections from Chandra and XMM-Newton been few and controversial. In the mean time, alternate methods of detecting the WHIM via far-UV absorbers are yielding impressive results. Roughly 100 highly-ionized metal ion detections (OVI, NV, etc) trace enriched, shock-heated gas at T<106 K. The catalog of far-UV WHIM absorbers is now large enough to not only detect the WHIM, but to start statistically-meaningful investigations as to its nature, origin, and distribution. Thermally-broadened Lyman-alpha absorbers are a second avenue of WHIM investigation which is similarly starting to produce results. Unlike either X-ray or UV metal line surveys, broad Lya studies are independent of metal enrichment and thus can trace hot gas in low-metallicity voids. Together, these two UV spectroscopic methods can account for 20% of the local baryons in the local universe.

  20. The use of ion exchange chromatography for demonstration of rubella-specific IgM antibodies.

    PubMed

    Nagy, G; Mezey, I

    1977-01-01

    IgM and IgG immunoglobulins of human sera were separated by stepwise column chromatography in QAE-Sephadex A-25 ion exchanger gel bed. The procedure resulted within 30 min in a fraction suitable for direct titration of rubella-specific IgM antibodies by haemagglutination inhibition test. The method proved to be a useful diagnostic tool for primary rubella. Serum samples of 13 individuals with previously acquired immunity, 152 patients with a recent rubella-like illness, and 194 pregnant women exposed to rubella infection were tested for the presence of rubella-specific IgM antibodies. Sera of individuals with previous immunity proved to be negative for specific IgM antibodies. Specific IgM titre was demonstrated in the blood of all the 25 patients with significant titre-rise tested because of rubella-like illness, and also in the sera of additional 8 patients whose serum samples were taken too late for demonstration of a rise in titre. Significant titre-rises were found in 5 women exposed to rubella infection, but only two of them exhibited rubella-specific IgM antibodies. The absence of specific IgM antibodies refers presumably to subclinical reinfection in the other three cases.

  1. A simple adherence test for detection of IgM antibodies in typhoid.

    PubMed

    Ong, L Y; Pang, T; Lim, S H; Tan, E L; Puthucheary, S D

    1989-07-01

    A simple adherence test to detect IgM antibodies in patients with typhoid is described. The test utilises the IgM-"capture" approach, in which the test serum is applied to microtitration plate wells previously coated with anti-human IgM, followed by application of a stained Salmonella typhi antigen suspension which shows adherence in positive cases. By this test, 58 (95%) of 61 sera from confirmed cases of typhoid possessed IgM antibodies to the H or O or both antigens of S. typhi. In patients for whom a diagnosis of typhoid was based only on a significant Widal-test titre, 31 (41%) of 76 sera had IgM antibodies to the H or O or both antigens of S. typhi. Some cross-reactivity of the IgM antibodies was detected, especially with the O antigens of S. paratyphi A and B. A total of 82 sera from non-typhoidal fevers (leptospirosis, typhus, dengue fever) showed no reactivity in this test. In normal sera there was no detectable IgM to the O antigen of S. typhi and only a small number (3.9%) had low levels of IgM to the H antigen. The significance and potential importance of this simple, sensitive, specific and economical test is discussed.

  2. Detection of IgM antibodies from cerebrospinal fluid and sera of dengue fever patients.

    PubMed

    Chen, W J; Hwang, K P; Fang, A H

    1991-12-01

    During the dengue epidemic from late 1987 to 1989, 6 specimens of cerebrospinal fluid (CSF) and sera for IgM detection were collected from 4 cases virologically confirmed dengue patients who had neural symptoms. Another 20 serum specimens, which had been diagnosed as dengue infection either virologically or serologically, were sent to the laboratory from Kaohsiung Medical College Hospital. All these specimens were also taken to detect the existence of IgM. The results showed that IgM could be detected from 14 out of 20 serum specimens. One of the positive specimens showed IgM can last up to 252 days after onset of illness. In addition, IgM was detected from both CSF and sera of all four dengue patients with neural symptoms. The IgM titer in CSF (less than or equal to 1:20) was always lower than that in serum (greater than or equal to 1:80). Two cases with sequentially collected specimens showed the fading of IgM titer in CSF. As a matter of fact, it became undetectable about a month after onset of illness, which is apparently different from the situation in serum.

  3. Biosynthesis of two forms of IgM heavy chains by normal mouse B lymphocytes. Membrane and secretory IgM.

    PubMed

    Vassalli, P; Tartakoff, A; Pink, J R; Jaton, J C

    1980-12-25

    A study of the biosynthesis of IgM by purified mouse spleen lymphocytes showed that these cells synthesize both 8 S membrane IgM and 19S secretory IgM, which is identical with plasma cell IgM except in its kinetics of processing, assembly, and secretion. The heavy (mu) chains of these two types of lymphocyte IgM differ in their ultimate fate, in processing, isoelectric point, and peptide composition. The separate precursors of the two mu chains have very similar mobilities in sodium dodecyl sulfate polyacrylamide gel electrophoresis, but they can be distinguished by the use of endoglucosaminidase H (endo-H) to remove core sugars, by two-dimensional electrophoresis, and by one-dimensional gel analysis in pulse-chase experiments. CNBr peptide patterns of intracellular "secretory" mu chains of lymphocytes and plasma cells were similar, but membrane mu chains had a COOH-terminal peptide different in structure from that of secreted mu chains, with a higher apparent molecular weight. PMID:6777384

  4. Expression and glycoengineering of functionally active heteromultimeric IgM in plants

    PubMed Central

    Loos, Andreas; Gruber, Clemens; Altmann, Friedrich; Mehofer, Ulrich; Hensel, Frank; Grandits, Melanie; Oostenbrink, Chris; Stadlmayr, Gerhard; Furtmüller, Paul G.; Steinkellner, Herta

    2014-01-01

    IgM antibodies are an important player of the human’s innate defense mechanisms and increasingly have gained interest as therapeutics. Although the expression of IgM antibodies in mammalian cell culture is established, this approach remains costly and alternative methods have not been developed yet. Plants have a proven record for the production of therapeutically relevant recombinant proteins. However, whether they are able to express proteins like IgM antibodies, which range among the most complex human proteins, remains unknown so far. Here we report the in planta generation of the functionally active monoclonal antitumor IgM PAT-SM6 (SM6). SM6 efficiently accumulates in plant leaves and assembles correctly into heterooligomers (pentamers and hexamers). Detailed glycosylation analysis exhibited complex and oligomannosidic N-glycans in a site-specific manner on human-serum IgM and on plant- and human-cell-line–produced SM6. Moreover, extensive in planta glycoengineering allowed the generation of SM6 decorated with sialylated human-type oligosaccharides, comparable to plasma-derived IgM. A glycosylated model of pentameric IgM exhibits different accessibility of the glycosylation sites, explaining site-specific glycosylation. Biochemical and biophysical properties and importantly biological activities of plant-derived SM6 glycoforms are comparable to the human-cell–derived counterparts. The in planta generation of one of the most complex human proteins opens new pathways toward the production of difficult-to-express proteins for pharmaceutical applications. Moreover, the generation of IgMs with a controlled glycosylation pattern allows the study of the so far unknown contribution of sugar moieties to the function of IgMs. PMID:24706782

  5. Transcriptional heterogeneity of IgM+ cells in rainbow trout (Oncorhynchus mykiss) tissues.

    PubMed

    Abós, Beatriz; Castro, Rosario; Pignatelli, Jaime; Luque, Alfonso; González, Lucia; Tafalla, Carolina

    2013-01-01

    TWO major classes of b lymphocytes have been described to date in rainbow trout: IgM(+) and IgT(+) cells. IgM(+) cells are mainly localized in the spleen, peripheral blood and kidney but are also found in other tissues. However, differences among IgM(+) cell populations attending to its location are poorly defined in fish. Thus, the aim of this work was to characterize the expression of different immune molecules such as chemokine receptors, Toll-like receptors (TLRs) and transcription factors on sorted IgM(+) lymphocytes from different rainbow trout tissues. IgM(+) populations from blood, spleen, kidney, gills, intestine and liver were isolated by cell sorting and the constitutive levels of transcription of these genes evaluated by real-time PCR. To further characterize B cells, we identified an MS4A sequence. In humans, the MS4A family includes several genes with immune functions, such as the B cell marker CD20 or FcRβ. Subsequently, we have also evaluated the mRNA levels of this MS4A gene in the different IgM(+) populations. The relevant differences in transcriptional patterns observed for each of these IgM(+) populations analyzed, point to the presence of functionally different tissue-specific B cell populations in rainbow trout. The data shown provides a pattern of genes transcribed in IgM(+) B cells not previously revealed in teleost fish. Furthermore, the constitutive expression of all the TLR genes analyzed in IgM(+) cells suggests an important role for these cells in innate immunity.

  6. Cytomegalovirus IgM Seroprevalence among Women of Reproductive Age in the United States.

    PubMed

    Wang, Chengbin; Dollard, Sheila C; Amin, Minal M; Bialek, Stephanie R

    2016-01-01

    Cytomegalovirus (CMV) IgM indicates recent active CMV infection. CMV IgM seroprevalence is a useful marker for prevalence of transmission. Using data from the National Health and Nutrition Examination Survey (NHANES) III 1988-1994, we present estimates of CMV IgM prevalence by race/ethnicity, provide a comparison of IgM seroprevalence among all women and among CMV IgG positive women, and explore factors possibly associated with IgM seroprevalence, including socioeconomic status and exposure to young children. There was no difference in IgM seroprevalence by race/ethnicity among all women (3.1%, 2.2%, and 1.6% for non-Hispanic white, non-Hispanic black and Mexican American, respectively; P = 0.11). CMV IgM seroprevalence decreased significantly with increasing age in non-Hispanic black women (P<0.001 for trend) and marginally among Mexican American women (P = 0.07), while no apparent trend with age was seen in non-Hispanic white women (P = 0.99). Among 4001 IgG+ women, 118 were IgM+, resulting in 4.9% IgM seroprevalence. In IgG+ women, IgM seroprevalence varied significantly by age (5.3%, 7.3%, and 3.7% for women of 12-19, 20-29, and 30-49 years; P = 0.04) and race/ethnicity (6.1%, 2.7%, and 2.0% for non-Hispanic white, non-Hispanic black, and Mexican American; P<0.001). The factors reported associated with IgG seroprevalence were not associated with IgM seroprevalence. The patterns of CMV IgM seroprevalence by age, race/ethnicity, and IgG serostatus may help understanding the epidemiology of congenital CMV infection as a consequence of vertical transmission and are useful for identifying target populations for intervention to reduce CMV transmission.

  7. Suppressor T cells prevent in vitro expression of IgM rheumatoid factor in some healthy adults

    SciTech Connect

    Koopman, W.J.

    1981-12-01

    Peripheral blood mononuclear leukocytes (MNL) from 11 of 30 healthy adults elaborated detectable IgM RF when stimulated with pokeweed mitogen. The influence of T cells on IgM RF production by autologous B Cells prepared from donors whose unfractionated MNL synthesized IgM RF in response to PMW was investigated. Untreated T cells supported IgM RF production by autologous B cells with optimal synthesis observed at T:B cell ratios of 2:1 at higher T:B cell ratios a decline in IgM RF production occurred. In contrast, at higher T:B cell ratios irradiated T cells supported consistently higher levels of IgM RF production than untreated T cells suggesting the presence of radiosensitive suppressor T cells for IgM RF in these individuals. Irridiated T cells were compared to untreated T cells for capacity to support IgM RF production by autologous B cells from 12 randomly selected donors at T:B cell ratios of 3:1. Untreated T cells from 4 of 12 individuals were capable of cooperating in induction of IgM RF production by autologous B cells, whereas irradiated T cells supported IgM RF production in 6 of 12 individuals. Levels of IgM RF production in all 6 individuals were significantly higher with irradiated T cells than with untreated T cells; in 2 individuals IgM RF synthesis by autologous B cells was observed only in the presence of irradiated T cells. In 4 of 6 individuals increases in the ratio of IgM RF total IgM synthesis occured with irradiated T cells (when compared to untreated T cells), suggesting disproportionate suppression of RF production. These results indicate the presence of radiosensitive T cells capable of suppressing IgM RF production in a significant fraction of healthy adults and raise the possibility that these cells may regulate in vivo expression of RF.

  8. Comparative Accuracy of the InBios Scrub Typhus Detect IgM Rapid Test for the Detection of IgM Antibodies by Using Conventional Serology.

    PubMed

    Kingston, Hugh W F; Blacksell, Stuart D; Tanganuchitcharnchai, Ampai; Laongnualpanich, Achara; Basnyat, Buddha; Day, Nicholas P J; Paris, Daniel H

    2015-10-01

    This study investigated the comparative accuracy of a recombinant 56-kDa type-specific antigen-based rapid diagnostic test (RDT) for scrub typhus for the detection of IgM antibodies by using conventional serology in well-characterized serum samples from undifferentiated febrile illness patients. The RDT showed high specificity and promising comparative accuracy, with 82% sensitivity and 98% specificity for samples defined positive at an IgM indirect immunofluorescence assay positivity cutoff titer of ≥1:1,600 versus 92% and 95% at ≥1:6,400, respectively.

  9. Circulating IgM Requires Plasma Membrane Disruption to Bind Apoptotic and Non-Apoptotic Nucleated Cells and Erythrocytes

    PubMed Central

    Hesketh, Emily E.; Dransfield, Ian; Kluth, David C.; Hughes, Jeremy

    2015-01-01

    Autoimmunity is associated with defective phagocytic clearance of apoptotic cells. IgM deficient mice exhibit an autoimmune phenotype consistent with a role for circulating IgM antibodies in apoptotic cell clearance. We have extensively characterised IgM binding to non-apoptotic and apoptotic mouse thymocytes and human Jurkat cells using flow cytometry, confocal imaging and electron microscopy. We demonstrate strong specific IgM binding to a subset of Annexin-V (AnnV)+PI (Propidium Iodide)+ apoptotic cells with disrupted cell membranes. Electron microscopy studies indicated that IgM+AnnV+PI+ apoptotic cells exhibited morphologically advanced apoptosis with marked plasma membrane disruption compared to IgM-AnnV+PI+ apoptotic cells, suggesting that access to intracellular epitopes is required for IgM to bind. Strong and comparable binding of IgM to permeabilised non-apoptotic and apoptotic cells suggests that IgM bound epitopes are 'apoptosis independent' such that IgM may bind any cell with profound disruption of cell plasma membrane integrity. In addition, permeabilised erythrocytes exhibited significant IgM binding thus supporting the importance of cell membrane epitopes. These data suggest that IgM may recognize and tag damaged nucleated cells or erythrocytes that exhibit significant cell membrane disruption. The role of IgM in vivo in conditions characterized by severe cell damage such as ischemic injury, sepsis and thrombotic microangiopathies merits further exploration. PMID:26121639

  10. Circulating IgM Requires Plasma Membrane Disruption to Bind Apoptotic and Non-Apoptotic Nucleated Cells and Erythrocytes.

    PubMed

    Hesketh, Emily E; Dransfield, Ian; Kluth, David C; Hughes, Jeremy

    2015-01-01

    Autoimmunity is associated with defective phagocytic clearance of apoptotic cells. IgM deficient mice exhibit an autoimmune phenotype consistent with a role for circulating IgM antibodies in apoptotic cell clearance. We have extensively characterised IgM binding to non-apoptotic and apoptotic mouse thymocytes and human Jurkat cells using flow cytometry, confocal imaging and electron microscopy. We demonstrate strong specific IgM binding to a subset of Annexin-V (AnnV)+PI (Propidium Iodide)+ apoptotic cells with disrupted cell membranes. Electron microscopy studies indicated that IgM+AnnV+PI+ apoptotic cells exhibited morphologically advanced apoptosis with marked plasma membrane disruption compared to IgM-AnnV+PI+ apoptotic cells, suggesting that access to intracellular epitopes is required for IgM to bind. Strong and comparable binding of IgM to permeabilised non-apoptotic and apoptotic cells suggests that IgM bound epitopes are 'apoptosis independent' such that IgM may bind any cell with profound disruption of cell plasma membrane integrity. In addition, permeabilised erythrocytes exhibited significant IgM binding thus supporting the importance of cell membrane epitopes. These data suggest that IgM may recognize and tag damaged nucleated cells or erythrocytes that exhibit significant cell membrane disruption. The role of IgM in vivo in conditions characterized by severe cell damage such as ischemic injury, sepsis and thrombotic microangiopathies merits further exploration.

  11. The role of IgM rheumatoid factor in experimental immune vasculitis.

    PubMed Central

    Floyd, M; Tesar, J T

    1979-01-01

    The effect of IgM rhematoid factor (RF) on reversepassive cutaneous Arthus reaction in rats was studied. The RF was obtained from the serum cryoglobulin of a patient with symptoms of purpura, arthralgia and digital gangrene. The cryoglobulins was of IgG-IgM type and when given i.v it induced a prompt hypocomplementaemia in experimental animals. The purified RF also induced low serum complement levels when injected i.v. along with complexes of non-complement-fixing, aggregated IgG. A reverse passive Arthus reaction was induced by intradermal injection of IgG anti-bovine serum albumin (BSA), followed by an i.v. dose of antigen (Ag). The cutaneous inflammatory reaction was aggravated by simultaneous administration of IgM RF intradermally, but not by IgM without antibody (Ab) properties. Intradermal injection of low concentrations of non-complement-fixing IgG anti-BSA, along with normal human IgM, followed by i.v. injection of BSA, resulted in a complete lack of cutaneous inflammation. At higher Ab concentrations there was only a mild inflammation. However, when IgM RF was substituted for normal IgM and injected with non-complement-fixing anti-BSA, an effective reverse passive cutaneous Arthus reaction and vasculitis was induced. The inflammatory response was greatly suppressed by decomplementation of animals by cobra venom factor. This study provides evidence favouring an inflammatory, complement-dependent role for RF in vasculitis. PMID:157238

  12. Physiological IgM Class Catalytic Antibodies Selective for Transthyretin Amyloid*

    PubMed Central

    Planque, Stephanie A.; Nishiyama, Yasuhiro; Hara, Mariko; Sonoda, Sari; Murphy, Sarah K.; Watanabe, Kenji; Mitsuda, Yukie; Brown, Eric L.; Massey, Richard J.; Primmer, Stanley R.; O'Nuallain, Brian; Paul, Sudhir

    2014-01-01

    Peptide bond-hydrolyzing catalytic antibodies (catabodies) could degrade toxic proteins, but acquired immunity principles have not provided evidence for beneficial catabodies. Transthyretin (TTR) forms misfolded β-sheet aggregates responsible for age-associated amyloidosis. We describe nucleophilic catabodies from healthy humans without amyloidosis that degraded misfolded TTR (misTTR) without reactivity to the physiological tetrameric TTR (phyTTR). IgM class B cell receptors specifically recognized the electrophilic analog of misTTR but not phyTTR. IgM but not IgG class antibodies hydrolyzed the particulate and soluble misTTR species. No misTTR-IgM binding was detected. The IgMs accounted for essentially all of the misTTR hydrolytic activity of unfractionated human serum. The IgMs did not degrade non-amyloidogenic, non-superantigenic proteins. Individual monoclonal IgMs (mIgMs) expressed variable misTTR hydrolytic rates and differing oligoreactivity directed to amyloid β peptide and microbial superantigen proteins. A subset of the mIgMs was monoreactive for misTTR. Excess misTTR was dissolved by a hydrolytic mIgM. The studies reveal a novel antibody property, the innate ability of IgMs to selectively degrade and dissolve toxic misTTR species as a first line immune function. PMID:24648510

  13. Platelet antibodies of the IgM class in immune thrombocytopenic purpura

    SciTech Connect

    Cines, D.B.; Wilson, S.B.; Tomaski, A.; Schreiber, A.D.

    1985-04-01

    The clinical course and response to therapy of patients with immune thrombocytopenic purpura (ITP) are not completely determined by the level of IgG present on the platelet surface. It is possible that antibodies of other immunoglobulin classes also play a role in platelet destruction in some of these patients. Therefore, the authors studied 175 patients with ITP for the presence of IgM anti-platelet antibodies using radiolabeled polyclonal or monoclonal anti-IgM. They observed that 57% of patients with clinical ITP had increased levels of IgM on their platelets, compared with normal controls and patients with thrombocytopenia who did not have ITP. They obtained similar results using either radiolabeled polyclonal or monoclonal anti-IgM, reagents whose integrity was first characterized using erythrocytes coated with defined amounts of IgM antibody. Among patients with increased platelet-IgM there was a significant correlation both with the presence of increased platelet-C3 as well as the amount of platelet-C3. The authors demonstrated the presence of warm-reacting IgM anti-platelet antibodies in the plasma of two of these patients who were further studied. These studies demonstrate the presence of warm-reacting IgM anti-platelet antibodies in some patients with ITP. They suggest that the binding of complement to platelets by IgM antibodies may initiate platelet clearance as well as enhance the effect of IgG antibodies in ITP.

  14. Antibody Binding Specificity for Kappa (Vκ) Light Chain-containing Human (IgM) Antibodies: Polysialic Acid (PSA) Attached to NCAM as a Case Study

    PubMed Central

    Watzlawik, Jens O.; Kahoud, Robert J.; Wootla, Bharath; Painter, Meghan M.; Warrington, Arthur E.; Carey, William A.; Rodriguez, Moses

    2016-01-01

    Antibodies of the IgM isotype are often neglected as potential therapeutics in human trials, animal models of human diseases as well as detecting agents in standard laboratory techniques. In contrast, several human IgMs demonstrated proof of efficacy in cancer models and models of CNS disorders including multiple sclerosis (MS) and amyotrophic lateral sclerosis (ALS). Reasons for their lack of consideration include difficulties to express, purify and stabilize IgM antibodies, challenge to identify (non-protein) antigens, low affinity binding and fundamental knowledge gaps in carbohydrate and lipid research. This manuscript uses HIgM12 as an example to provide a detailed protocol to detect antigens by Western blotting, immunoprecipitations and immunocytochemistry. HIgM12 targets polysialic acid (PSA) attached to the neural cell adhesion molecule (NCAM). Early postnatal mouse brain tissue from wild type (WT) and NCAM knockout (KO) mice lacking the three major central nervous system (CNS) splice variants NCAM180, 140 and 120 was used to evaluate the importance of NCAM for binding to HIgM12. Further enzymatic digestion of CNS tissue and cultured CNS cells using endoneuraminidases led us to identify PSA as the specific binding epitope for HIgM12. PMID:27404858

  15. A natural human IgM that binds to gangliosides is therapeutic in murine models of amyotrophic lateral sclerosis

    PubMed Central

    Xu, Xiaohua; Denic, Aleksandar; Jordan, Luke R.; Wittenberg, Nathan J.; Warrington, Arthur E.; Wootla, Bharath; Papke, Louisa M.; Zoecklein, Laurie J.; Yoo, Daehan; Shaver, Jonah; Oh, Sang-Hyun; Pease, Larry R.; Rodriguez, Moses

    2015-01-01

    ABSTRACT Amyotrophic lateral sclerosis (ALS) is a devastating, fatal neurological disease that primarily affects spinal cord anterior horn cells and their axons for which there is no treatment. Here we report the use of a recombinant natural human IgM that binds to the surface of neurons and supports neurite extension, rHIgM12, as a therapeutic strategy in murine models of human ALS. A single 200 µg intraperitoneal dose of rHIgM12 increases survival in two independent genetic-based mutant SOD1 mouse strains (SOD1G86R and SOD1G93A) by 8 and 10 days, delays the onset of neurological deficits by 16 days, delays the onset of weight loss by 5 days, and preserves spinal cord axons and anterior horn neurons. Immuno-overlay of thin layer chromatography and surface plasmon resonance show that rHIgM12 binds with high affinity to the complex gangliosides GD1a and GT1b. Addition of rHIgM12 to neurons in culture increases α-tubulin tyrosination levels, suggesting an alteration of microtubule dynamics. We previously reported that a single peripheral dose of rHIgM12 preserved neurological function in a murine model of demyelination with axon loss. Because rHIgM12 improves three different models of neurological disease, we propose that the IgM might act late in the cascade of neuronal stress and/or death by a broad mechanism. PMID:26035393

  16. A natural human IgM that binds to gangliosides is therapeutic in murine models of amyotrophic lateral sclerosis.

    PubMed

    Xu, Xiaohua; Denic, Aleksandar; Jordan, Luke R; Wittenberg, Nathan J; Warrington, Arthur E; Wootla, Bharath; Papke, Louisa M; Zoecklein, Laurie J; Yoo, Daehan; Shaver, Jonah; Oh, Sang-Hyun; Pease, Larry R; Rodriguez, Moses

    2015-08-01

    Amyotrophic lateral sclerosis (ALS) is a devastating, fatal neurological disease that primarily affects spinal cord anterior horn cells and their axons for which there is no treatment. Here we report the use of a recombinant natural human IgM that binds to the surface of neurons and supports neurite extension, rHIgM12, as a therapeutic strategy in murine models of human ALS. A single 200 µg intraperitoneal dose of rHIgM12 increases survival in two independent genetic-based mutant SOD1 mouse strains (SOD1G86R and SOD1G93A) by 8 and 10 days, delays the onset of neurological deficits by 16 days, delays the onset of weight loss by 5 days, and preserves spinal cord axons and anterior horn neurons. Immuno-overlay of thin layer chromatography and surface plasmon resonance show that rHIgM12 binds with high affinity to the complex gangliosides GD1a and GT1b. Addition of rHIgM12 to neurons in culture increases α-tubulin tyrosination levels, suggesting an alteration of microtubule dynamics. We previously reported that a single peripheral dose of rHIgM12 preserved neurological function in a murine model of demyelination with axon loss. Because rHIgM12 improves three different models of neurological disease, we propose that the IgM might act late in the cascade of neuronal stress and/or death by a broad mechanism.

  17. [Relevance of the new test Access Toxo IgM (II) in the serological interpretation of toxoplasmosis in pregnancy].

    PubMed

    Flori, P; Hafid, J; Raberin, H; Patural, H; Varlet, M N; Tran Manh Sung, R

    2002-01-01

    The detection of specific IgM is an important element in the diagnosis of seroconversion. In this study, we compared the new test Access Toxo IgM (II) versus the previous test Access Toxo IgM (I). This comparative study was done with 507 random sera and 89 selected sera (seroconversion, residual IgM, non specific IgM). The sensitivity and the specificity are higher with the new test (100% & 100% versus 97% & 98,6% respectively). Then, the high sensitivity of Access Toxo IgM (II) is more convenient for the earlier diagnosis of primary Toxoplasma infection. Moreover, the Access Toxo IgM (II) test was evaluated with our first results in routine laboratory (11,000 serology results between June 2000 and June 2001). With these results, we can confirm the specificity of the new test, and also determine the relevance of anti-toxoplasmic IgM. Moreover, an increase of sensitivity leads to an increase of residual IgM detection: 64% (37/58) of positive IgM is residual (> 4 months) as compared to 35% (27/78) with the previous test. In these case, the initiation of other tests such as the avidity test is necessary to validate or invalidate a seroconversion during pregnancy.

  18. Site-Specific N-Glycosylation of Recombinant Pentameric and Hexameric Human IgM

    NASA Astrophysics Data System (ADS)

    Moh, Edward S. X.; Lin, Chi-Hung; Thaysen-Andersen, Morten; Packer, Nicolle H.

    2016-07-01

    Glycosylation is known to play an important role in IgG antibody structure and function. Polymeric IgM, the largest known antibody in humans, displays five potential N-glycosylation sites on each heavy chain monomer. IgM can exist as a pentamer with a connecting singly N-glycosylated J-chain (with a total of 51 glycosylation sites) or as a hexamer (60 glycosylation sites). In this study, the N-glycosylation of recombinant pentameric and hexameric IgM produced by the same human cell type and culture conditions was site-specifically profiled by RP-LC-CID/ETD-MS/MS using HILIC-enriched tryptic and GluC glycopeptides. The occupancy of all putative N-glycosylation sites on the pentameric and hexameric IgM were able to be determined. Distinct glycosylation differences were observed between each of the five N-linked sites on the IgM heavy chains. While Asn171, Asn332, and Asn395 all had predominantly complex type glycans, differences in glycan branching and sialylation were observed between the sites. Asn563, a high mannose-rich glycosylation site that locates in the center of the IgM polymer, was only approximately 60% occupied in both the pentameric and hexameric IgM forms, with a difference in relative abundance of the glycan structures between the pentamer and hexamer. This study highlights the information obtained by characterization of the site-heterogeneity of a highly glycosylated protein of high molecular mass with quaternary structure, revealing differences that would not be seen by global glycan or deglycosylated peptide profiling.

  19. Simultaneous Quantification of Anticardiolipin IgG and IgM by Time Resolved Fluoroimmunoassay

    PubMed Central

    Liu, Jie; Li, Mei; Ye, Yan; Chen, Yu

    2016-01-01

    The autoimmune disease antiphospholipid syndrome (APS) is characterized by the presence of anticardiolipin antibodies (aCL), along with anti-β2-glycoprotein I (β2GPI) antibodies and lupus anticoagulant (LA). In this study, we developed a time-resolved fluoroimmunoassay (TRFIA) system for simultaneous quantification of aCL IgG and IgM. A 96-well microtiter plate precoated with the complex of cardiolipin from bovine heart and bovine β2GPI was incubated with the anticardiolipin IgG and IgM standard substance or serum, and the conjugate of Eu3+-labeled anti-human IgG and Sm3+-labeled anti-human IgM was pipetted to the wells to form a tipical double-antibody-sandwich immunoreactions; finally the fluorescent intensity of Eu3+ and Sm3+ was detected to reflect the quantity of anticardiolipin IgG and IgM. This assay showed a good relationship between fluorescence intensities and the concentration of anticardiolipin antibody(aCL) IgG and IgM, with a low-end sensitivity of 0.1 U/ml for IgG and 0.1 U/ml for IgM, respectively. The intra- and inter-assay coefficients of variation (CV) of the calibrators was 3.0% and 4.51% for IgG, and 2.76% and 4.45% for IgM. The average recovery was 100.38% for aCL IgG and 100.45% for aCL IgM. For serum samples, the results of our method showed a good correlation with those obtained with ELISA kit. Simultaneous detection of aCL-IgG and aCL-IgM in the same reaction well can optimize assay performance by avoiding potential influence of different reaction conditions-timing, and well-to-well difference in concentration and characteristics of cardiolipin antigen. The results of a combo aCL-IgG and aCL-IgM assay for the same sample are more consistent and more reliable. This dual-label time-resolved fluoroimmunoassay is sensitive for detecting aCL IgG and IgM across a wide concentration range with stable reagents and may assist in the clinical diagnosis of antiphospholipid syndrome. PMID:27661084

  20. Distribution of radiolabeled human and mouse monoclonal IgM antibodies in murine models.

    PubMed

    Halpern, S E; Hagan, P L; Chen, A; Birdwell, C R; Bartholomew, R M; Burnett, K G; David, G S; Poggenburg, K; Merchant, B; Carlo, D J

    1988-10-01

    The distribution and kinetics of six human and one murine monoclonal IgM antibodies (MoAb) were studied in BALB/c mice. Labeling was with 111In, 75Se, and 125I. The monomers and pentamers of certain MoAbs were studied. Human distribution studies were also performed. The serum containing [111In]MoAb was obtained from one of the patients 24 hr after administration and injected into mice which were then killed and assayed for 111In distribution. In general, the [75Se] and [111In]MoAbs had distribution and kinetic patterns that were similar while the 125I-labeled MoAbs dehalogenated after 4 hr. Monomers and pentamers had highly similar distributions suggesting that the distribution of IgMs may be based on factors other than molecular size. The murine IgM showed a somewhat different distribution in mice than did human IgMs. Serum from the patient containing [111In]MoAb had a distribution in mice similar to that of the patient with high liver and gastrointestinal uptake. The human imaging indicates that it is possible to target tumor with human IgM MoAbs, but significant problems remain in regard to their clinical use.

  1. Circulating microparticles carry oxidation-specific epitopes and are recognized by natural IgM antibodies.

    PubMed

    Tsiantoulas, Dimitrios; Perkmann, Thomas; Afonyushkin, Taras; Mangold, Andreas; Prohaska, Thomas A; Papac-Milicevic, Nikolina; Millischer, Vincent; Bartel, Caroline; Hörkkö, Sohvi; Boulanger, Chantal M; Tsimikas, Sotirios; Fischer, Michael B; Witztum, Joseph L; Lang, Irene M; Binder, Christoph J

    2015-02-01

    Oxidation-specific epitopes (OSEs) present on apoptotic cells and oxidized low density lipoprotein (OxLDL) represent danger-associated molecular patterns that are recognized by different arcs of innate immunity, including natural IgM antibodies. Here, we investigated whether circulating microparticles (MPs), which are small membrane vesicles released by apoptotic or activated cells, are physiological carriers of OSEs. OSEs on circulating MPs isolated from healthy donors and patients with ST-segment elevation myocardial infarction (STE-MI) were characterized by flow cytometry using a panel of OSE-specific monoclonal antibodies. We found that a subset of MPs carry OSEs on their surface, predominantly malondialdehyde (MDA) epitopes. Consistent with this, a majority of IgM antibodies bound on the surface of circulating MPs were found to have specificity for MDA-modified LDL. Moreover, we show that MPs can stimulate THP-1 (human acute monocytic leukemia cell line) and human primary monocytes to produce interleukin 8, which can be inhibited by a monoclonal IgM with specificity for MDA epitopes. Finally, we show that MDA(+) MPs are elevated at the culprit lesion site of patients with STE-MI. Our results identify a subset of OSE(+) MPs that are bound by OxLDL-specific IgM. These findings demonstrate a novel mechanism by which anti-OxLDL IgM antibodies could mediate protective functions in CVD.

  2. Influenza Virus-Specific Neutralizing IgM Antibodies Persist for a Lifetime

    PubMed Central

    Skountzou, Ioanna; Satyabhama, Lakshmipriyadarshini; Stavropoulou, Anastasia; Ashraf, Zuhha; Esser, E. Stein; Vassilieva, Elena; Koutsonanos, Dimitrios; Compans, Richard

    2014-01-01

    Detection of immunoglobulin M (IgM) antibodies has long been used as an important diagnostic tool for identifying active viral infections, but their relevance in later stages has not been clearly defined in vivo. In this study, we followed the kinetics, longevity, and function of influenza virus-specific IgM antibodies for 2 years following sublethal infection of mice with live mouse-adapted A/PR/8/34 virus or immunization with formalin-inactivated virus. These groups mounted robust protective immune responses and survived lethal challenges with 50× 50% lethal dose (LD50) mouse-adapted A/PR/8/34 virus 600 days after the primary exposure. Surprisingly, the virus-specific IgM antibodies persisted along with IgG antibodies, and we found a significantly higher number of IgM-positive (IgM+) virus-specific plasma cells than IgG+ plasma cells that persisted for at least 9 months postexposure. The IgM antibodies were functional as they neutralized influenza virus in the presence of complement just as well as IgG antibodies did. PMID:25165027

  3. Roles of heavy and light chains in IgM polymerization.

    PubMed Central

    Bornemann, K D; Brewer, J W; Beck-Engeser, G B; Corley, R B; Haas, I G; Jäck, H M

    1995-01-01

    IgM antibodies are secreted as multisubunit polymers that consist of as many as three discrete polypeptides: mu heavy chains, light (L) chains, and joining (J) chains. We wished to determine whether L chains that are required to confer secretory competence on immunoglobulin molecules must be present for IgM to polymerize--that is, for intersubunit disulfide bonds to form between mu chains. Using a L-chain-loss variant of an IgM-secreting hybridoma, we demonstrated that mu chains were efficiently polymerized independent of L chains, in a manner similar to that observed for conventional microL complexes, and that the mu polymers incorporated J chain. These mu polymers were not secreted but remained associated with the endoplasmic reticulum-resident chaperone BiP (GRP78). This finding is consistent with the endoplasmic reticulum being the subcellular site of IgM polymerization. We conclude that mu chain alone has the potential to direct the polymerization of secreted IgM, a process necessary but not sufficient for IgM to attain secretory competence. Images Fig. 1 Fig. 2 Fig. 3 Fig. 4 PMID:7761423

  4. Distribution of radiolabeled human and mouse monoclonal IgM antibodies in murine models

    SciTech Connect

    Halpern, S.E.; Hagan, P.L.; Chen, A.; Birdwell, C.R.; Bartholomew, R.M.; Burnett, K.G.; David, G.S.; Poggenburg, K.; Merchant, B.; Carlo, D.J.

    1988-10-01

    The distribution and kinetics of six human and one murine monoclonal IgM antibodies (MoAb) were studied in BALB/c mice. Labeling was with /sup 111/In, /sup 75/Se, and /sup 125/I. The monomers and pentamers of certain MoAbs were studied. Human distribution studies were also performed. The serum containing (/sup 111/In)MoAb was obtained from one of the patients 24 hr after administration and injected into mice which were then killed and assayed for /sup 111/In distribution. In general, the (/sup 75/Se) and (/sup 111/In)MoAbs had distribution and kinetic patterns that were similar while the /sup 125/I-labeled MoAbs dehalogenated after 4 hr. Monomers and pentamers had highly similar distributions suggesting that the distribution of IgMs may be based on factors other than molecular size. The murine IgM showed a somewhat different distribution in mice than did human IgMs. Serum from the patient containing (/sup 111/In)MoAb had a distribution in mice similar to that of the patient with high liver and gastrointestinal uptake. The human imaging indicates that it is possible to target tumor with human IgM MoAbs, but significant problems remain in regard to their clinical use.

  5. IgM nephropathy: is it closer to minimal change disease or to focal segmental glomerulosclerosis?

    PubMed

    Brugnano, R; Del Sordo, R; Covarelli, C; Gnappi, E; Pasquali, S

    2016-08-01

    Immunoglobulin (Ig)M nephropathy (IgMN), known since 1978, is a very controversial clinicopathological entity characterized by IgM diffuse deposits in the mesangium at immunofluorescence whereas light microscop identifies minimal glomerular lesion, hypercellularity and expansion of the mesangium or sclerotic focal, segmental lesion. Clinically, it is a nephrotic syndrome, especially in pediatric patients, or asymptomatic proteinuria and/or isolated hematuria. These characteristics narrowly define IgMN between minimal change disease and focal segmental glomerulosclerosis, so it is not often recognized as a separate pathology. Homogeneous epidemiologic, pathogenetic, clinical or histological data are not available. Recent research on the pathogenetic role of mesangial IgM has, however, renewed interest in IgMN and naturally the controversies.

  6. IgM–IgG cryoglobulinaemia with IgM paraprotein component

    PubMed Central

    Klein, F.; van Rood, J. J.; van Furth, R.; Radema, H.

    1968-01-01

    Four patients with mixed IgM–IgG cryoglobulinaemia are described. Clinically they all had some features of an autoimmune disease, while two of them had a lympho-epithelial tumour in the parotid gland. The mixed cryoglobulins of all patients contained an IgM paraprotein with the properties of a rheumatoid factor. They can be regarded as cryoprecipitates of a rheumatoid factor with autologous IgG. In one case the parotid tumour, and not the bone marrow, produced the IgM paraprotein. The clinical significance of the cryoglobulins is discussed. The IgM paraproteins with rheumatoid factor activity may be an expression of an underlying abnormality of the immunological system of these patients. ImagesFig. 1Fig. 2Fig. 4Fig. 5Fig. 6Fig. 7 PMID:5701952

  7. Reversal of IgM deficiency following a gluten-free diet in seronegative celiac disease.

    PubMed

    Montenegro, Lucia; Piscitelli, Domenico; Giorgio, Floriana; Covelli, Claudia; Fiore, Maria Grazia; Losurdo, Giuseppe; Iannone, Andrea; Ierardi, Enzo; Di Leo, Alfredo; Principi, Mariabeatrice

    2014-12-14

    Selective IgM deficiency (sIGMD) is very rare; it may be associated with celiac disease (CD). We present the case of an 18-year-old man with sIGMD masking seronegative CD. Symptoms included abdominal pain, diarrhea and weight loss. Laboratory tests showed reduced IgM, DQ2-HLA and negative anti-transglutaminase. Villous atrophy and diffuse immature lymphocytes were observed at histology. Tissue transglutaminase mRNA mucosal levels showed a 6-fold increase. The patient was treated with a gluten-free diet (GFD) and six months later the symptoms had disappeared, the villous architecture was restored and mucosal tissue transglutaminase mRNA was comparable to that of healthy subjects. After 1 year of GFD, a complete restoration of normal IgM values was observed and duodenal biopsy showed a reduction of immature lymphocytes and normal appearance of mature immune cells.

  8. Symptomatic Secondary Selective IgM Immunodeficiency in Adult Man with Undiagnosed Celiac Disease

    PubMed Central

    Magen, Eli; Feldman, Viktor; Joseph, Mishal; Israel, Hadari

    2012-01-01

    Selective IgM immunodeficiency (SIgMID) is a heterogeneous disorder with no known genetic background and may occur as a primary or a secondary condition. Celiac disease has been reported in association with several humeral immunodeficiencies, including isolated severe selective IgA deficiency, panhypogammaglobulinemia, and isolated combined IgA and IgM deficiency. There are only few reported cases of pediatric and adult patients with SIgMID and celiac disease. In this paper, we describe an adult patient with a symptomatic secondary SIgMID associated with undiagnosed celiac disease, with a resolution of clinical symptoms of immunodeficiency and serum IgM normalization following a gluten-free diet. PMID:25374731

  9. Diagnostic and prognostic significance of the IgM antibody to the Hepatitis delta virus

    SciTech Connect

    Farci, P.; Gerin, J.L.; Aragona, M.; Lindsey, I.; Crivelli, O.; Balestrieri, A.; Smedile, A.; Thomas, H.C.; Rizzetto, M.

    1986-03-21

    The IgM class antibody to the hepatitis delta virus (HDV) was determined in different clinical categories of hepatitis B surface antigen carriers infected by the HDV (positive in the test for total antibody to HDV). The IgM antibody was found at high titers in each 70 patients with inflammatory liver disease and at a low titer in one six patients with inactive cirrhosis; it was not found in eight carriers with normal liver histology. Testing for Igm antibody to HDV distinguishes hepatitis B surface antigen carriers who have underlying inflammatory HDV liver disease from those with past HDV infection and provides prognostic information on the course of chronic HDV hepatitis.

  10. Suppression of IgM Proteolysis by Conformational Stabilization Through Excipients

    PubMed Central

    Mueller, Monika; Loh, Maybelle Q. T.; Gagnon, Pete

    2015-01-01

    Protease activity from host cell lines may cause product loss or affect the quality of recombinant proteins. In this study, we showed that excipients like glycine and sorbitol reduce the proteolysis of an immunoglobulin M (IgM) in the presence of added proteases like α-chymotrypsin, papain, and pepsin. The activity of the proteases in the IgM-protective environments was conserved or even enhanced as tested using low molecular weight substrates. Thus, a higher resistance against proteolytic degradation appears to be caused by the conformational stabilization of the IgM due to preferential exclusion of sorbitol and glycine. PMID:26839826

  11. Levels of natural IgM antibodies against phosphorylcholine in healthy individuals and in patients undergoing isolated limb perfusion.

    PubMed

    Padilla, Niubel Diaz; Ciurana, Caroline; van Oers, Joep; Ogilvie, Aernout C; Hack, C Erik

    2004-10-01

    Natural IgM antibodies against phosphorylcholine (anti-Pc IgM) resemble C-reactive protein (CRP) regarding specificity and have gained increasing attention because of their supposed role in clearance of damaged cells and in cardiovascular disease. In order to quantify these antibodies in human plasma, we have developed an ELISA system, in which p-aminophenylphosphorylcholine (PCH) coupled to human serum albumin (HSA) was coated on microtiters plates. Human plasma or serum samples were incubated in the plates, after which bound anti-Pc IgM was detected with mouse anti-human IgM-HRP. Pre-incubation of plasma with competitors such as phosphorylcholine, phosphorylethanolamine, phosphorylserine or glycine-HSA, confirmed that the ELISA was specific for anti PC IgM. Levels of anti Pc IgM in a cohort of healthy donors differed by more than 100-fold, whereas the fluctuation of anti-Pc IgM levels in individuals over time was small (coefficient of variation between 6% to 25%). Furthermore, there was no correlation between CRP and anti-Pc IgM in this cohort. Levels of anti-Pc IgM in the normal donors correlated significantly with IgM binding to apoptotic cells. To test the hypothesis that anti-Pc IgM can bind to neo-antigens expressed on necrotic or apoptotic cells, anti-Pc IgM was also quantified in patients with tumors undergoing isolated limb perfusion with tumor necrosis factor-alpha (TNF-alpha). Following this procedure, a significant decrease of circulating anti-Pc IgM relative to total IgM was found in all five patients tested. In conclusion, we have developed a specific and reproducible ELISA for anti Pc IgM quantification. Fluctuation of levels of these natural antibodies over time in healthy individuals was limited, although the variation among individuals was large. Significant decreases of levels of anti-Pc IgM were found to occur during tissue damage.

  12. Selective IgM deficiency in adults: phenotypically and functionally altered profiles of peripheral blood lymphocytes.

    PubMed Central

    Ohno, T; Inaba, M; Kuribayashi, K; Masuda, T; Kanoh, T; Uchino, H

    1987-01-01

    Peripheral blood lymphocytes from four patients with selective IgM deficiency were examined phenotypically and functionally. Although B cell subpopulations determined by surface immunoglobulins were within normal or nearly normal range, T8+ cells were significantly increased and T4/T8 ratios were inverted in three patients. IgM specific hyporesponsiveness in the PWM-driven immunoglobulin production system was observed in all four patients. Ia-like antigen positive T cells were increased in two patients; both had increased Leu2a+ Leu15+ suppressor-effector cells. In addition, Leu3a+ Leu8+ suppressor-inducer cells were increased in one of these two patients. Excessive (either IgM-specific or isotype non-specific) suppressor activity of T cells and IgM specific hyporesponsiveness of non-T cells were observed in these two patients in the recombination plaque assay. Although these results showed the complexity of the pathogenesis of this syndrome, they suggested that suppressor-associated T cells may play a role in some patients with selective IgM deficiency. PMID:2958191

  13. Granular IgM Deposition at Basement Membrane Zone in an Infant with Diffuse Cutaneous Mastocytosis.

    PubMed

    Kumudhini, Subramanian; Rao, Raghavendra; Salgaonkar, Gauri; Shetty, Sricharith; Pai, Sathish

    2016-01-01

    Diffuse Cutaneous mastocytosis (DCM) occurs due to abnormal accumulation of mast cells in the skin. We report an 8-month-old infant presented papulovesicular lesions, predominantly on the trunk. Skin biopsy revealed subepidermal bulla, interspersed with mast cells, eosinophils and neutrophils. Direct immunofluorescence microscopy of perilesional skin revealed nonspecific deposition of IgM in granular pattern along the dermoepidermal junction. PMID:27688466

  14. Granular IgM Deposition at Basement Membrane Zone in an Infant with Diffuse Cutaneous Mastocytosis

    PubMed Central

    Kumudhini, Subramanian; Rao, Raghavendra; Salgaonkar, Gauri; Shetty, Sricharith; Pai, Sathish

    2016-01-01

    Diffuse Cutaneous mastocytosis (DCM) occurs due to abnormal accumulation of mast cells in the skin. We report an 8-month-old infant presented papulovesicular lesions, predominantly on the trunk. Skin biopsy revealed subepidermal bulla, interspersed with mast cells, eosinophils and neutrophils. Direct immunofluorescence microscopy of perilesional skin revealed nonspecific deposition of IgM in granular pattern along the dermoepidermal junction. PMID:27688466

  15. Baryonic Content in the Warm-Hot IGM at Low Redshift

    NASA Technical Reports Server (NTRS)

    Sonneborn, George; Shull, M.; Danforth, C.; Moos, W.

    2007-01-01

    Baryons are 4.5% of the universe's mass/energy density; only 10% of these are in stars, galaxies, and clusters. At low-redshift 90% of baryons are in the IGM, 30% in Ly-alpha forest, but most are in hot gas (10(exp 5-7) K) produced by shocks during structure formation. O VI 1032-38 A are the best tracers of this gas. The distribution of O VI absorbers observed by FUSE rises as N(sup -2+/-0.2, down to 10(exp 13)/sq cm. Integrated to logN=13, 7% of baryons reside in the O VI-bearing IGM at 10% solar metallicity, T approx. 10(exp 5.5) K. At redshift z<0.1 metals have been transported less than 800/h kpc from L* galaxies and 200/h kpc from 0.1 L* galaxies. The steepness of dN/dz means that low-N absorbers contribute an equal mass of hot IGM as higher N gas. The total mass of O VI-bearing gas in the IGM depends on determining the turnover in dN/dz at low N(O VI). Future observations by FUSE are needed to reach lower N and to reduce the uncertainty in the dN/dz power law.

  16. Serological survey of anti-group A rotavirus IgM in UK adults.

    PubMed Central

    Cox, M. J.; Medley, G. F.

    2003-01-01

    Rotaviral associated disease of infants in the UK is seasonal and infection in adults not uncommon but the relationship between these has been little explored. Adult sera collected monthly for one year from routine hospital samples were screened for the presence of anti-group A rotavirus immunoglobulin M class antibodies as a marker of recent infection. Anti-rotavirus IgM was seen in all age groups throughout the year with little obvious seasonal variation in the distribution of antibody levels. IgM concentrations and the proportion seropositive above a threshold both increased with age with high concentrations consistently observed in the elderly. Results suggest either high infection rates of rotavirus in adults, irrespective of seasonal disease incidence in infants, IgM persistence or IgM cross-reactivity. These results support recent evidence of differences between infant and adult rotavirus epidemiology and highlight the need for more extensive surveys to investigate age and time related infection and transmission of rotavirus. PMID:12948372

  17. Performance of Elecsys toxo IgG and IgM immunoassays.

    PubMed

    Van Helden, Josef

    2009-01-01

    Roche Diagnostics has developed two new rapid and fully automated assays for the detection of Anti-Toxo-IgG and Toxo-IgM antibodies from human sera and plasma. The performance evaluation of the Elecsys Toxo IgG resulted in a sensitivity of 100% and a specificity of 99.91% showing that this assay allows a very sensitive and specific detection of Toxo IgG antibodies with an excellent discrimination of positive and negative results. The performance evaluation of the Elecsys Toxo IgM assay revealed a sensitivity of 100% and a specificity of 99.11% after resolution. Less reactivity towards persistent Toxo IgM antibodies in samples from Toxoplasma infections > 3 months was found with Elecsys Toxo IgM. The performance evaluation data demonstrate that the Elecsys Toxo IgG and Elecsys Toxo IgM assays are reliable tools in routine diagnostics of Toxoplasma infections with the additional advantage of a high throughput on fully automated analyzers MODULAR ANALYTICS Serum Work Area.

  18. Granular IgM Deposition at Basement Membrane Zone in an Infant with Diffuse Cutaneous Mastocytosis

    PubMed Central

    Kumudhini, Subramanian; Rao, Raghavendra; Salgaonkar, Gauri; Shetty, Sricharith; Pai, Sathish

    2016-01-01

    Diffuse Cutaneous mastocytosis (DCM) occurs due to abnormal accumulation of mast cells in the skin. We report an 8-month-old infant presented papulovesicular lesions, predominantly on the trunk. Skin biopsy revealed subepidermal bulla, interspersed with mast cells, eosinophils and neutrophils. Direct immunofluorescence microscopy of perilesional skin revealed nonspecific deposition of IgM in granular pattern along the dermoepidermal junction.

  19. Natural IgM antibodies: the orphaned molecules in immune surveillance.

    PubMed

    Vollmers, H Peter; Brändlein, Stephanie

    2006-08-01

    Natural IgM antibodies are typical victims of prejudices which originated in the mid 80 s. Over the years, these molecules were considered as the pariahs among the immune competent molecules and their characteristic properties, like low affinity, cross-reactivity and pentameric structure, were assessed as useless, difficult, nebulous, etc. Today, mainly based on a few scientists' persistent work and the key discoveries on innate immune recognition, natural IgM antibodies are "back on stage". Their role in the immune response against bacteria, viruses, fungi and possibly modified self-components as well as in therapy and diagnosis of malignancies is accepted. All the so far negatively judged features are seen in a different light, e.g. low affinity seems to be good for function and does not exclude specificity, and cross-reactivity is no longer judged as unspecific, but instead as a very economic way of immune recognition. And at last, with the use of natural IgM antibodies, a new field of tumor-specific targets has been encountered, the carbo-neo-epitopes. Therefore, by having learned from nature, the renaissance of natural IgM antibodies opens a new area of cancer therapeutics and diagnostics. PMID:16820243

  20. B-1 B cell IgM antibody initiates T cell elicitation of contact sensitivity.

    PubMed

    Askenase, P W; Tsuji, R F

    2000-01-01

    Although B-1 B cells have received considerable attention, their actual role in the normal functioning of the immune system is unclear. The hypothesized role of B-1 cell IgM in natural protective immunity is just being established. We have uncovered a separate and novel role for B-1 cell IgM in initiating the elicitation of acquired T cell-dependent contact sensitivity (CS), the prototype of in vivo T cell immunity, early after immunization (within 4 days). The recent recognition of a similarly unanticipated role of B cells in a variety of T cell responses, may indicate that B-1 cell IgM has a broader role in immunity than thought previously. We showed that 24 hr CS responses, and rises in local IFN-gamma levels at 24 hrs later after antigen (Ag) challenge the ears, were absent in pan B cell and antibody deficient mice. The mechanism of B cell involvement in CS-initiation is via local C5a generation early (1-2 hrs) after antigen (Ag) challenge of the ears, in 4 day contact sensitized mice. C5a activates local mast cells to release serotonin (5-HT) and TNF alpha to induce endothelial ICAM-1 and VCAM-1, leading to T cell recruitment. We hypothesized that C5a was generated via complement activation due to antibodies forming local AgAb complexes, and that B-1 cell IgM was involved because isotype switching of B-2 cells to produce C-activating IgG isotypes, could not occur as early as day 4. Indeed, B-1 cell deficient CBA/N-xid mice lacked C5a in 2 hr ear extracts, and had impaired CS ear swelling and elaboration of IFN-gamma at 24 hrs. Importantly, adoptive transfer of purified normal peritoneal B-1 cells, or just i.v. injection of Ag-specific IgM monoclonal antibodies in sensitized xid, restored deficient early C5a and late 24 hr ear swelling. These results suggest that early after Ag challenge, specific B-1 cell IgM, produced at distant sites by prior sensitization, forms AgAb complexes that trigger elaboration of C5a, to activate mast cell release of vasoactive TNF

  1. Placebo-controlled trial of rituximab in IgM anti-myelin–associated glycoprotein neuropathy

    PubMed Central

    Viala, Karine; Nicolas, Guillaume; Créange, Alain; Vallat, Jean-Michel; Pouget, Jean; Clavelou, Pierre; Vial, Christophe; Steck, Andreas; Musset, Lucile; Marin, Benoit

    2013-01-01

    Objective: To determine whether rituximab 375 mg/m2 was efficacious in patients with immunoglobulin M (IgM) anti-myelin–associated glycoprotein antibody demyelinating neuropathy (IgM anti-MAG demyelinating neuropathy). Methods: Fifty-four patients with IgM anti-MAG demyelinating neuropathy were enrolled in this randomized, double-blind, placebo-controlled trial. The inclusion criteria were inflammatory neuropathy cause and treatment (INCAT) sensory score (ISS) ≥4 and visual analog pain scale >4 or ataxia score ≥2. The primary outcome was mean change in ISS at 12 months. Results: Twenty-six patients were randomized to a group receiving 4 weekly infusions of 375 mg/m2 rituximab, and 28 patients to placebo. Intention-to-treat analysis, with imputation of missing ISS values by the last observation carried forward method, showed a lack of mean change in ISS at 12 months, 1.0 ± 2.7 in the rituximab group, and 1.0 ± 2.8 in the placebo group. However, changes were observed, in per protocol analysis at 12 months, for the number of patients with an improvement of at least 2 points in the INCAT disability scale (p = 0.027), the self-evaluation scale (p = 0.016), and 2 subscores of the Short Form–36 questionnaire. Conclusions: Although primary outcome measures provide no evidence to support the use of rituximab in IgM anti-MAG demyelinating neuropathy, there were improvements in several secondary outcomes in per protocol analysis. Level of evidence: This study provides Class I evidence that rituximab is ineffective in improving ISS in patients with IgM anti-MAG demyelinating neuropathy. PMID:23667063

  2. APPRAISAL OF PRENATAL ANTI-TOXOPLASMA GONDII (IGG+IGM)- IHA/IGM-ELISA SCREENING IN SINGLE SAMPLES VIA IGG AVIDITY TEST.

    PubMed

    El-Bali, Mohammed; Zaglool, Dina A M; Khodari, Yousif A W; Al-Harthi, Saeed A

    2016-04-01

    Congenital toxoplasmosis is associated with important morbidity and mortality. Since vertical transmission of Toxoplasma gondii can occur in acute cases, antenatal screening for recent infections is vital. Accurate determination of acute toxoplasmosis requires a combination of immunoassays, usually not routinely applied for screening purposes. This study evaluated the anti-T. gondii (IgG+IgM)/IgM prenatal screening procedure by IgG avidity assay. The routine prenatal screening for (IgG+IgM) anti-T. gondii by indirect hemagglutination (IHA) in serum samples was done of 2247 pregnant women who attended two hospitals between 2011 and 2013 revealed 487 (21.7%) positive samples. Examination of IHA-positive sera by IgM and IgG/IgG-avidity concurrent ELISA tests revealed 7 positive and 3 border-line IgM-ELISA titers during the initial check-up of 10 women, who were then followed up at 3-4 week-intervals. Among these, 4 (40%) showed simultaneous high avidity IgG antibodies, indicating distant infection by the parasite, and no anti-T. gondii specific IgG could be detected in follow-up sera of two cases (20%), indicating false IgM initial positive results. Only 4 (40%) women showed simultaneous IgM and low avidity IgG antibodies indicating active infections. Avoidance of an over-diagnosis of acute toxoplasmosis Anti-T. gondii (IgG+IgM)/IgM prenatal screening must be supplemented by a discriminative test like IgG avidity ELISA. PMID:27363056

  3. Optimal Cutoff and Accuracy of an IgM Enzyme-Linked Immunosorbent Assay for Diagnosis of Acute Scrub Typhus in Northern Thailand: an Alternative Reference Method to the IgM Immunofluorescence Assay.

    PubMed

    Blacksell, Stuart D; Lim, Cherry; Tanganuchitcharnchai, Ampai; Jintaworn, Suthatip; Kantipong, Pacharee; Richards, Allen L; Paris, Daniel H; Limmathurotsakul, Direk; Day, Nicholas P J

    2016-06-01

    The enzyme-linked immunosorbent assay (ELISA) has been proposed as an alternative serologic diagnostic test to the indirect immunofluorescence assay (IFA) for scrub typhus. Here, we systematically determine the optimal sample dilution and cutoff optical density (OD) and estimate the accuracy of IgM ELISA using Bayesian latent class models (LCMs). Data from 135 patients with undifferentiated fever were reevaluated using Bayesian LCMs. Every patient was evaluated for the presence of an eschar and tested with a blood culture for Orientia tsutsugamushi, three different PCR assays, and an IgM IFA. The IgM ELISA was performed for every sample at sample dilutions from 1:100 to 1:102,400 using crude whole-cell antigens of the Karp, Kato, and Gilliam strains of O. tsutsugamushi developed by the Naval Medical Research Center. We used Bayesian LCMs to generate unbiased receiver operating characteristic curves and found that the sample dilution of 1:400 was optimal for the IgM ELISA. With the optimal cutoff OD of 1.474 at a sample dilution of 1:400, the IgM ELISA had a sensitivity of 85.7% (95% credible interval [CrI], 77.4% to 86.7%) and a specificity of 98.1% (95% CrI, 97.2% to 100%) using paired samples. For the ELISA, the OD could be determined objectively and quickly, in contrast to the reading of IFA slides, which was both subjective and labor-intensive. The IgM ELISA for scrub typhus has high diagnostic accuracy and is less subjective than the IgM IFA. We suggest that the IgM ELISA may be used as an alternative reference test to the IgM IFA for the serological diagnosis of scrub typhus.

  4. Optimal Cutoff and Accuracy of an IgM Enzyme-Linked Immunosorbent Assay for Diagnosis of Acute Scrub Typhus in Northern Thailand: an Alternative Reference Method to the IgM Immunofluorescence Assay

    PubMed Central

    Blacksell, Stuart D.; Tanganuchitcharnchai, Ampai; Jintaworn, Suthatip; Kantipong, Pacharee; Richards, Allen L.; Day, Nicholas P. J.

    2016-01-01

    The enzyme-linked immunosorbent assay (ELISA) has been proposed as an alternative serologic diagnostic test to the indirect immunofluorescence assay (IFA) for scrub typhus. Here, we systematically determine the optimal sample dilution and cutoff optical density (OD) and estimate the accuracy of IgM ELISA using Bayesian latent class models (LCMs). Data from 135 patients with undifferentiated fever were reevaluated using Bayesian LCMs. Every patient was evaluated for the presence of an eschar and tested with a blood culture for Orientia tsutsugamushi, three different PCR assays, and an IgM IFA. The IgM ELISA was performed for every sample at sample dilutions from 1:100 to 1:102,400 using crude whole-cell antigens of the Karp, Kato, and Gilliam strains of O. tsutsugamushi developed by the Naval Medical Research Center. We used Bayesian LCMs to generate unbiased receiver operating characteristic curves and found that the sample dilution of 1:400 was optimal for the IgM ELISA. With the optimal cutoff OD of 1.474 at a sample dilution of 1:400, the IgM ELISA had a sensitivity of 85.7% (95% credible interval [CrI], 77.4% to 86.7%) and a specificity of 98.1% (95% CrI, 97.2% to 100%) using paired samples. For the ELISA, the OD could be determined objectively and quickly, in contrast to the reading of IFA slides, which was both subjective and labor-intensive. The IgM ELISA for scrub typhus has high diagnostic accuracy and is less subjective than the IgM IFA. We suggest that the IgM ELISA may be used as an alternative reference test to the IgM IFA for the serological diagnosis of scrub typhus. PMID:27008880

  5. Enzyme immunoassays for IgG and IgM antibodies to Toxoplasma gondii based on enhanced chemiluminescence.

    PubMed Central

    Crouch, C F

    1995-01-01

    AIMS--To evaluate the clinical performance of enzyme immunoassays for IgG and IgM antibodies to Toxoplasma gondii based on enhanced chemiluminescence. METHODS--Classification of routine clinical samples from the originating laboratories was compared with that obtained using the chemiluminescence based assays. Resolution of discordant results was achieved by testing in alternative enzyme immunoassays (IgM) or by an independent laboratory using the dye test (IgG). RESULTS--Compared with resolved data, the IgM assay was found to be highly specific (100%) with a cut off selected to give optimal performance with respect to both the early detection of specific IgM and the detection of persistent levels of specific IgM (sensitivity 98%). Compared with resolved data, the IgG assay was shown to have a sensitivity and a specificity of 99.4%. CONCLUSIONS--The Amerlite Toxo IgM assay possesses high levels of sensitivity and specificity. Assay interference due to rheumatoid factor like substances is not a problem. The Amerlite Toxo IgG assay possesses good sensitivity and specificity, but is less sensitive for the detection of seroconversion than methods detecting both IgG and IgM. PMID:7560174

  6. A selective defect in IgM antigen receptor synthesis and transport causes loss of cell surface IgM expression on tolerant B lymphocytes.

    PubMed Central

    Bell, S E; Goodnow, C C

    1994-01-01

    To explore the biochemical basis for maintaining immunological tolerance by functional inactivation of self-reactive B lymphocytes, transgenic mice carrying rearranged anti-lysozyme immunoglobulin transgenes and a lysozyme transgene were used as a source of large numbers of tolerant self-reactive B cells. Antigen receptors of the IgD isotype were expressed at normal levels on tolerant B cells, contained the heterodimeric MB1/B29 signalling component of the receptor complex and were structurally indistinguishable from IgD on nontolerant B cells. In contrast, cell surface expression of IgM receptor complexes on tolerant B cells was greatly reduced, despite normal expression of mRNA encoding the receptor components. Three-fold fewer immunoreactive mu heavy chains were detectable after a short period of biosynthetic labelling and the immunoreactive mu chains produced were paired with kappa light chains and assembled normally into intact receptor complexes containing the MB1/B29 heterodimer. Nascent IgM receptor complexes nevertheless failed to be processed into an endoglycosidase H-resistant form in the tolerant B cells and thus appeared to be selectively blocked in their transport from the endoplasmic reticulum to the medial Golgi. These findings demonstrate that intracellular trafficking of antigen receptor complexes is regulated by exposure to receptor stimuli at the cell surface causing a long-lasting decrease in surface receptor expression on tolerant B cells. Images PMID:8112296

  7. Evidence for hydrophobic region within heavy chains of mouse B lymphocyte membrane-bound IgM

    PubMed Central

    Vassalli, Pierre; Tedghi, Rachel; Lisowska-Bernstein, Barbara; Tartakoff, Alan; Jaton, Jean-Claude

    1979-01-01

    The gel filtration behavior, in the presence of detergents, of membrane-bound IgM from normal mouse spleen B lymphocytes was compared to that of secretory IgM from mouse plasma cells. The proteins were labeled either by surface radioiodination or biosynthetically with radioactive amino acids. Cell lysates were fractionated on calibrated Sepharose 6B columns in the presence of the detergents Nonidet P-40 or deoxycholate. Eluted fractions were immunoprecipitated and the reduced or unreduced precipitates were analyzed by sodium dodecyl sulfate gel electrophoresis followed by radioautography. Surface 125I-labeled 8S IgM exhibited a gel filtration pattern in Nonidet P-40 corresponding to much higher apparent molecular weight than that of secretory 8S IgM, a difference that almost disappeared when gel filtration was performed in the presence of deoxycholate, which forms much smaller micelles than does Nonidet P-40. Biosynthetically labeled lymphocytes contain two types of IgM molecules differing in their gel filtration behavior and fate: one identical to secretory 8S IgM of plasma cells and secreted in the medium during chase periods, and the other identical to surface 125I-labeled IgM and remaining cell-associated. Because the surface-bound 8S IgM was not found to be associated with other labeled molecules, it is likely that the detergent-binding behavior of surface IgM is due to a hydrophobic segment carried by these Ig molecules. That lymphocytes synthesize two types of μ chains was also shown by the use of tunicamycin, an inhibitor of glycosylation. In its presence, two unglycosylated μ chains were observed: one identical in size to that made by tunicamycin-treated plasma cells, and the second slightly larger. Gel filtration in Nonidet P-40 of the cell lysates of tunicamycin-treated lymphocytes showed that the nonsecretory 8S IgM contains this second type of μ chains, whereas the IgM molecules of the secretory type contain plasma cell-like μ chains. It is

  8. Circulating monoclonal IgM lambda cryoglobulin with collagen type I affinity in vasculitis.

    PubMed Central

    Clemmensen, I; Jensen, B A; Hølund, B; Kappelgaard, E; Neilsen, H

    1986-01-01

    A previously fit 66-years-old male primarily presented symptoms compatible with Henock-Schönlein's purpura, from which he seemingly recovered. Shortly hereafter he relapsed with an IgM lambda essential monoclonal cryoglobulinemia type I, presenting a systemic, necrotizing vasculitis, with low titer of circulating immune complexes and complement consumption. Glucocorticoid treatment and plasmapheresis did not prevent an ultimately lethal course. An indirect immunoperoxidase technique showed that the cryo-IgM bound to the interstitial connective tissue corresponding to the localization of collagen type I. In addition it bound to affinity purified human procollagen type I. These results indicate, that the IgM lambda of the proband was an autoantibody with collagen type I specificity. Images Fig. 1 Fig. 2 Fig. 3 Fig. 4 Fig. 5 PMID:3098468

  9. Blood-nerve barrier in IgM paraproteinemic neuropathy: a clinicopathologic assessment.

    PubMed

    Kanda, T; Usui, S; Beppu, H; Miyamoto, K; Yamawaki, M; Oda, M

    1998-02-01

    We report the pathologic findings in a patient with sensorimotor neuropathy associated with Waldenström's macroglobulinemia, particularly in relation to blood-nerve barrier defects. The monoclonal IgM was of kappa type and possessed anti-HNK-1 activity. A sural nerve biopsy specimen revealed severe loss of myelinated and unmyelinated nerve fibers and gaps between adjacent endothelial cells of small endoneurial vessels. Postmortem findings 3 years later included severe loss of myelinated nerve fibers and diffuse infiltration by lymphoplasmacytic B cells throughout the peripheral nervous system, sparing the central nervous system. Findings in this case suggest an immune attack against endoneurial endothelial cells with permeation of IgM into peripheral nerve tissue. PMID:9498055

  10. Normal human serum contains a natural IgM antibody cytotoxic for human neuroblastoma cells.

    PubMed Central

    Ollert, M W; David, K; Schmitt, C; Hauenschild, A; Bredehorst, R; Erttmann, R; Vogel, C W

    1996-01-01

    Neuroblastoma (NB) is characterized by the second highest spontaneous regression of any human malignant disorder, a phenomenon that remains to be elucidated. In this study, a survey of 94 normal human adult sera revealed a considerable natural humoral cytotoxicity against human NB cell lines in approximately one-third of the tested sera of both genders. Specific cell killing by these sera was in the range of 40% to 95%. Serum cytotoxicity was dependent on an intact classical pathway of complement. By several lines of evidence, IgM antibodies were identified as the cytotoxic factor in the sera. Further analyses revealed that a 260-kDa protein was recognized by natural IgM of cytotoxic sera in Western blots of NB cell extracts. The antigen was expressed on the surface of seven human NB cell lines but not on human melanoma or other control tumor cell lines derived from kidney, pancreas, colon, bone, skeletal muscle, lymphatic system, and bone marrow. Furthermore, no reactivity was observed with normal human fibroblasts, melanocytes, and epidermal keratinocytes. The antigen was expressed in vivo as detected by immunohistochemistry in both the tumor of a NB patient and NB tumors established in nude rats from human NB cell lines. Most interestingly, the IgM anti-NB antibody was absent from the sera of 11 human NB patients with active disease. The anti-NB IgM also could not be detected in tumor tissue obtained from a NB patient. Collectively, our data suggest the existence of a natural humoral immunological tumor defense mechanism, which could account for the in vivo phenomenon of spontaneous NB tumor regression. Images Fig. 1 Fig. 3 Fig. 4 Fig. 5 Fig. 6 PMID:8633097

  11. [Use of staphylococcal protein A for the detection of rubella virus IgM antibodies].

    PubMed

    Crovari, P; Gasparini, R; Bono, A; Lusian, F; Tassi, G C

    1979-11-30

    The ability of Staphylococcus aureus protein A to bind human serum immunoglobulin of different classes and the conditions which allow a selective reduction of IgG in comparison to other immunoglobulin classes were preliminarly studied. Working under standardized conditions, a recovery of 3.5% of the initial value for IgG, of 40-50% for IgM and of 70-75% for IgA was obtained. Rubella HI antibodies, before and after staphylococcus treatment, were titred in 88 sera collected from adult women at different times from hexantema and in 210 sera of 109 subjects exposed to eventual contamination. The results were compared with the ones obtained by means of sucrose density gradient centrifugation followed by titration of rubella HI antibodies in the fractions. The tests proved that HI antibodies titration, before and after serum treatment with staphylococcal protein A, can be used, as presumptive test, for the research of rubella specific IgM antibodies. In fact, all the sera, on which the presence of rubella IgM antibodies was proved by sucrose density gradient centrifugation, were also positive after staphylococcus treatment. In 15% of sera resulting positive for HI rubella antibodies after staphylococcus treatment, usually at low titer, the presence of specific IgM antibodies was not detected by serum fractionation on sucrose density gradient ("false positive" cases). The results of serological diagnosis, carried out by using treatment with staphylococcal protein A and serum fractionation on sucrose density gradient, matched the clinical-epidemiological data. Twenty-seven women, out of 28 examined during the first three months of pregnancy and for whom serological diagnosis of a "recent" rubella virus infection was negative, had normal children at normal time. Therefore only in one case a spontaneous abortion occurred for uncertain causes. Rubella virus was isolated from the embryo of a woman who interrupted pregnancy because asymptomatic rubella virus infection was

  12. Chlamydia trachomatis IgM seropositivity during pregnancy and assessment of its risk factors.

    PubMed

    Rahman, M; Chowdhury, S B; Akhtar, N; Jahan, M; Jahan, M K; Jebunnahar, S

    2014-01-01

    The study was undertaken to determine socio-demographic and reproductive risk factors associated with Chlamydia trachomaties IgM seropositivity during pregnancy. This cross sectional comparative study was carried out in the obstetrics outdoor of Bangabandhu Sheikh Mujib Medical University (BSMMU), Dhaka, Bangladesh in collaboration with the department of Virology between the periods from July 2007 to December 2008. Pregnant women at their first visit to the hospital were approached consecutively and asked to complete a questionnaire and 2cc blood was collected from each subject for Chlamydia trachomatis IgM antibody testing using ELISA method. The study population was divided into two groups according to the presence and absence of serum Chlamydia trachomatis IgM antibody. Finally socio-demographic and reproductive risk factors were compared between the groups. Among 172 women the sero-prevalence of Chlamydia IgM was 41%. The multiple logistic regression model (step wise) finally extracted for characteristics correlated with seropositivity. Ten years or less (≤SSC) education (OR 2.6 95% CI 1.1to 5.9), history of adverse pregnancy outcome (OR 2.8 95% CI 1.2 to 6.5) and multiple sex partner of husband (OR 4.1 95% CI 1.2 to 14.8) were associated with chlamydia infection. The use of condom (OR 0.28 95% CI 0.12 to 0.63) was associated with decreased risk of infection. Chlamydia trachomatis infection during pregnancy is associated with risk factors on the basis of which selective screening can be done.

  13. Role of membrane-bound IgM in Trypanosoma cruzi evasion from immune clearance.

    PubMed

    Garcia, I E; Lima, M R; Marinho, C R; Kipnis, T L; Furtado, G C; Alvarez, J M

    1997-04-01

    We have recently described that Trypanosoma cruzi parasites of the reticulotropic Y strain increase their resistance to antibody-induced clearance during their interaction with the vertebrate host immune system. In the present study, we observed that trypomastigotes of the myotropic CL strain isolated from normal host also display an increased resistance to immune clearance when compared to parasites obtained from immunosuppressed donors. Through fluorescence-activated cell sorting analysis, we have observed that the high expression of membrane-bound IgM antibodies on Y and CL trypomastigotes correlates with their enhanced resistance to Ig-induced clearance. Trypomastigotes from normal mice were essentially refractory to the in vitro binding of immunoglobulins, showing that their membrane structures were completely covered by IgM antibodies. These findings suggest that this isotype does not efficiently mediate immune clearance. Moreover, membrane-bound IgM antibodies limited the amount of IgG attached to the parasite and, as a consequence, impaired efficient immune clearance. Through this mechanism, trypomastigotes of T. cruzi could increase their persistence in the bloodstream thus favoring parasite transmission to its hematophagous host vector in the early acute phase of the disease.

  14. Monoclonal IgM antibody exhibiting high-affinity binding and cryoglobulin properties.

    PubMed Central

    Ballard, D W; Kranz, D M; Voss, E W

    1983-01-01

    A monoclonal IgM antibody (18-2-3) derived from cell fusion of (NZB X NZW) F1 splenocytes following secondary immunization with fluorescein-conjugated keyhole limpet hemocyanin was shown to exhibit high intrinsic binding affinity and cryoinsolubility. Affinity-purified preparations were determined to be IgM by immunochemical, electrophoretic, and chromatographic analyses. An intrinsic association constant (Ka) of 2.9 X 10(10) M-1 (at 2 degrees C) was measured by first-order dissociation-rate analysis. Antibody solubility at low concentration (approximately equal to 50 micrograms/ml) was shown, by absorption spectroscopy, to be temperature dependent between 4 degrees C and 32 degrees C. Insolubility at low temperature (4 degrees C) was reversible in the presence of homologous fluorescyl hapten, indicative of active site involvement in the mechanism of cryoglobulin-18-2-3 complex formation. Characteristics of clone 18-2-3 are discussed in terms of (i) its potential use as a model for examining the mechanism of cryoprecipitation and (ii) the proposed relationship between affinity maturation and the IgM to IgG class switch. Images PMID:6348779

  15. IgM multiple myeloma with an extremely rare non-aggressive presentation: A case report

    PubMed Central

    Greuter, Thomas; Browne, Martin; Dommann-Scherrer, Corina; Binder, Daniel; Renner, Christoph; Kapp, Ursula

    2016-01-01

    In the present study, the case of a 41-year-old man with immunoglobulin (Ig)M multiple myeloma (MM) that presented with an unusually non-aggressive clinical course who has survived for >9 years to date, is presented. Initial diagnosis of symptomatic MM was established according to the International Myeloma Working Group consensus statement and guidelines. Due to the mild symptoms, no therapy was administered and the patient was closely followed up. Eight years after initial diagnosis, clinical, morphological and genetic progression occurred with the development of hypercalcemia, progressively deteriorating polyneuropathy, clonal expansion of plasma cells up to 50% of hematopoietic cells and demonstration of the typical t(11;14) translocation (Ig heavy chain locus rearrangement). Subsequently, 4 cycles of induction chemotherapy with velcade, cyclophosphamide and dexamethasone, were administered. At the time of writing, the patient remained alive in generally good health. To the best of our knowledge, with a survival time of >9 years, this case reports the longest survival time of an IgM MM patient to date, which contradicts previous evidence that suggests IgM MM exhibits an aggressive clinical course.

  16. Pathogenic relevance of IgG and IgM antibodies against desmoglein 3 in blister formation in pemphigus vulgaris.

    PubMed

    Tsunoda, Kazuyuki; Ota, Takayuki; Saito, Masataka; Hata, Tsuyoshi; Shimizu, Atsushi; Ishiko, Akira; Yamada, Taketo; Nakagawa, Taneaki; Kowalczyk, Andrew P; Amagai, Masayuki

    2011-08-01

    Pemphigus vulgaris is an autoimmune disease caused by IgG antibodies against desmoglein 3 (Dsg3). Previously, we isolated a pathogenic mAb against Dsg3, AK23 IgG, which induces a pemphigus vulgaris-like phenotype characterized by blister formation. In the present study, we generated a transgenic mouse expressing AK23 IgM to examine B-cell tolerance and the pathogenic role of IgM. Autoreactive transgenic B cells were found in the spleen and lymph nodes, whereas anti-Dsg3 AK23 IgM was detected in the cardiovascular circulation. The transgenic mice did not develop an obvious pemphigus vulgaris phenotype, however, even though an excess of AK23 IgM was passively transferred to neonatal mice. Similarly, when hybridoma cells producing AK23 IgM were inoculated into adult mice, no blistering was observed. Immunoelectron microscopy revealed IgM binding at the edges of desmosomes or interdesmosomal cell membranes, but not in the desmosome core, where AK23 IgG binding has been frequently detected. Furthermore, in an in vitro dissociation assay using cultured keratinocytes, AK23 IgG and AK23 IgM F(ab')(2) fragments, but not AK23 IgM, induced fragmentation of epidermal sheets. Together, these observations indicate that antibodies must gain access to Dsg3 integrated within desmosomes to induce the loss of keratinocyte cell-cell adhesion. These findings provide an important framework for improved understanding of B-cell tolerance and the pathophysiology of blister formation in pemphigus.

  17. 7S IgG rheumatoid factor and hidden 19S IgM rheumatoid factor in juvenile chronic arthritis.

    PubMed

    Roizenblatt, S; Goldenberg, J; Gabriel, A; Hilário, M O; Atra, E

    1993-01-01

    Hidden 19S IgM rheumatoid factor (RF) and 7S IgG RF were assayed in 24 children with juvenile chronic arthritis (JCA) and 26 controls, in whole sera and in the IgM and IgG containing fractions of the serum after separation by acid gel filtration. When non-fractioned serum was used, IgM RF detection was positive in 4.2% by latex fixation test and 4.2% by ELISA. IgG RF was positive in 91.7% by ELISA. In the IgM containing fraction of the serum, the positivity for IgM RF was 41.7% by LFT and 54.2% by ELISA. In the IgG containing fraction the positivity for IgG RF was 100%. These results suggest toward the need of previous serum dissociation before hidden 19S IgM RF is assayed and demonstrate high prevalence of 7S IgG RF.

  18. Ontogeny of B cells expressing IgM in embryonic and larval tissues of the American grass frog, Rana pipiens.

    PubMed

    Zettergren, L D

    2000-06-01

    Affinity-purified, fluorochrome-tagged F(ab')(2) antibody fragments specific for heavy (mu) chains of Rana pipiens IgM were prepared from hyperimmune rabbit sera. By using two-color immunofluorescent procedures we observed that (1) the first cells expressing IgM, termed pre-B cells, lack detectable quantities of membrane or surface IgM but contain detectable quantities of cytoplasmic IgM (smu(-)/cmu(+)), (2) sIgM(+) B cells were the second type of IgM containing cell to appear in development, and (3) plasma cells, which contain copious quantities of cIgM, were the final phenotype to appear in the development of B cells expressing IgM. These cells were first observed in the pronephros of the developing urogenital system. Shortly after their appearance in the pronephros, cells in B lineages were observed in the liver. These observations (1) are consistent with recent studies of B lymphopoiesis in the aorta-gonad-mesonephros (AGM) region in endothermic vertebrates, including mice, (2) suggest that there are fundamental ontogenetic and phylogenetic similarities between cells and tissues of developing vertebrate immune systems, and (3) evoke questions concerning the possible function(s) of lymphocytes in developing anurans up to metamorphosis and beyond.

  19. Somatically Hypermutated Plasmodium-Specific IgM(+) Memory B Cells Are Rapid, Plastic, Early Responders upon Malaria Rechallenge.

    PubMed

    Krishnamurty, Akshay T; Thouvenel, Christopher D; Portugal, Silvia; Keitany, Gladys J; Kim, Karen S; Holder, Anthony; Crompton, Peter D; Rawlings, David J; Pepper, Marion

    2016-08-16

    Humoral immunity consists of pre-existing antibodies expressed by long-lived plasma cells and rapidly reactive memory B cells (MBC). Recent studies of MBC development and function after protein immunization have uncovered significant MBC heterogeneity. To clarify functional roles for distinct MBC subsets during malaria infection, we generated tetramers that identify Plasmodium-specific MBCs in both humans and mice. Long-lived murine Plasmodium-specific MBCs consisted of three populations: somatically hypermutated immunoglobulin M(+) (IgM(+)) and IgG(+) MBC subsets and an unmutated IgD(+) MBC population. Rechallenge experiments revealed that high affinity, somatically hypermutated Plasmodium-specific IgM(+) MBCs proliferated and gave rise to antibody-secreting cells that dominated the early secondary response to parasite rechallenge. IgM(+) MBCs also gave rise to T cell-dependent IgM(+) and IgG(+)B220(+)CD138(+) plasmablasts or T cell-independent B220(-)CD138(+) IgM(+) plasma cells. Thus, even in competition with IgG(+) MBCs, IgM(+) MBCs are rapid, plastic, early responders to a secondary Plasmodium rechallenge and should be targeted by vaccine strategies. PMID:27473412

  20. Role of Natural Autoantibodies and Natural IgM Anti-Leucocyte Autoantibodies in Health and Disease

    PubMed Central

    Lobo, Peter Isaac

    2016-01-01

    We review how polyreactive natural IgM autoantibodies (IgM-NAA) protect the host from invading micro-organisms and host neo-antigens that are constantly being produced by oxidation mechanisms and cell apoptosis. Second, we discuss how IgM-NAA and IgM anti-leukocyte antibodies (IgM-ALA) inhibits autoimmune inflammation by anti-idiotypic mechanisms, enhancing removal of apoptotic cells, masking neo-antigens, and regulating the function of dendritic cells (DC) and effector cells. Third, we review how natural IgM prevents autoimmune disorders arising from pathogenic IgG autoantibodies, triggered by genetic mechanisms (e.g., SLE) or micro-organisms, as well as by autoreactive B and T cells that have escaped tolerance mechanisms. Studies in IgM knockout mice have clearly demonstrated that regulatory B and T cells require IgM to effectively regulate inflammation mediated by innate, adaptive, and autoimmune mechanisms. It is, therefore, not surprising why the host positively selects such autoreactive B1 cells that generate IgM-NAA, which are also evolutionarily conserved. Fourth, we show that IgM-ALA levels and their repertoire can vary in normal humans and disease states and this variation may partly explain the observed differences in the inflammatory response after infection, ischemic injury, or after a transplant. We also show how protective IgM-NAA can be rendered pathogenic under non-physiological conditions. We also review IgG-NAA that are more abundant than IgM-NAA in plasma. However, we need to understand if the (Fab)2 region of IgG-NAA has physiological relevance in non-disease states, as in plasma, their functional activity is blocked by IgM-NAA having anti-idiotypic activity. Some IgG-NAA are produced by B2 cells that have escaped tolerance mechanisms and we show how such pathogenic IgG-NAA are regulated to prevent autoimmune disease. The Fc region of IgG-NAA can influence inflammation and B cell function in vivo by binding to activating and inhibitory Fc

  1. IgG and IgM autoantibody differences in discoid and systemic lupus patients

    PubMed Central

    Chong, Benjamin F.; Tseng, Lin-chiang; Lee, Thomas; Vasquez, Rebecca; Li, Quan Z.; Zhang, Song; Karp, David R.; Olsen, Nancy J.; Mohan, Chandra

    2012-01-01

    Systemic lupus (SLE) patients with discoid lupus (DLE) were reported to have milder disease. To test this observation, we employed sandwich arrays containing 98 autoantigens to compare autoantibody profiles of SLE subjects without DLE (DLE−SLE+) (N=9), SLE subjects with DLE (DLE+SLE+) (N=10), DLE subjects without SLE (DLE+SLE−) (N=11), and healthy controls (N=11). We validated differentially expressed autoantibodies using immunoassays in DLE−SLE+ (N=18), DLE+SLE+ (N=17), DLE+SLE− (N=23), and healthy subjects (N=22). Arrays showed 15 IgG autoantibodies (ten against nuclear antigens) and four IgM autoantibodies that were differentially expressed (q-value<0.05). DLE−SLE+ subjects had higher IgG autoantibodies against dsDNA, ssDNA, dsRNA, histone H2A and H2B, and SS-A (52 kDa) than all other groups including DLE+SLE+ subjects (p<0.05). Immunoassays measuring anti-dsDNA, -ssDNA, and -SS-A (52 kDa) IgG autoantibodies showed similar trends (p<0.05). Healthy and DLE+SLE−subjects expressed higher IgM autoantibodies against alpha beta crystallin, lipopolysaccharide, heat shock cognate 70, and desmoglein-3 than DLE+SLE+ and DLE−SLE+ subjects. IgG:IgM ratios of autoantibodies against nuclear antigens progressively rose from healthy to DLE−SLE+ subjects. In conclusion, lower IgG autoantibodies against nuclear antigens in DLE+SLE+ versus DLE−SLE+ subjects suggest that DLE indicates lower disease severity. Higher IgM autoantibodies against selected antigens in healthy and DLE+SLE−subjects may be non-pathogenic. PMID:22763789

  2. Production and characterization of monoclonal antibodies to IgM of Pacific herring (Clupea pallasii)

    USGS Publications Warehouse

    Purcell, Maureen K.; Bromage, Erin S.; Silva, Jessica; Hansen, John D.; Badil, Samantha M.; Woodson, James C.; Hershberger, Paul K.

    2012-01-01

    Pacific herring (Clupea pallasii) have a central role in the North Pacific ecosystem as a forage fish species and are natural reservoirs of several important finfish pathogens, including Viral hemorrhagic septicemia virus (VHSV). Here, we report the identification of the gene encoding the immunoglobulin mu (IgM) heavy chain, as well as the development and characterization of monoclonal antibodies (MAbs) that specifically react with Pacific herring IgM. Pacific herring immunoglobulin was purified and consisted of heavy and light chains of approximately 80 and 25 kDa. Three hybridoma clones were initially identified by ELISA as reactive with purified immunoglobulin but only one clone was able to detect an 80 kDa protein in Pacific and Atlantic herring (Clupea harengus) whole plasma by denaturing western blot. However, all three MAbs were able to precipitate an 80 kDa protein from Pacific herring and LCMS sequencing of peptide fragments derived from this protein matched the predicted amino acid sequence of the cloned, heavy chain gene. In addition, two of the MAbs stained cells within the putative lymphocyte gates for the spleen, anterior kidney and posterior kidney but were not reactive for myeloid/granulocyte gates, which is consistent with these MAbs reacting with surface IgM+ B-cells. To our knowledge, this is the first report of IgM-related gene sequences and anti-IgM monoclonal antibodies from any member of the family Clupeidae. The antibodies produced in this study are critical for achieving our long-term goal of conducting serological surveillance to assess pathogen exposure in natural populations of Pacific herring.

  3. HOW TO SEARCH FOR ISLANDS OF NEUTRAL HYDROGEN IN THE z ∼ 5.5 IGM

    SciTech Connect

    Malloy, Matthew; Lidz, Adam

    2015-02-01

    Observations of the Lyman-alpha (Lyα) forest may allow reionization to complete as late as z ∼ 5.5, provided the ionization state of the intergalactic medium (IGM) is sufficiently inhomogeneous at these redshifts. In this case, significantly neutral islands may remain among highly ionized gas with the ionized regions allowing some transmission through the Lyα forest. This possibility has the important virtue that it is eminently testable with existing Lyα forest data. In particular, we describe three observable signatures of significantly neutral gas in the z ∼ 5.5 IGM. We use mock quasar spectra produced from numerical simulations of reionization to develop these tests. First, we quantify how the abundance and length of absorbed regions in the forest increase with the volume-averaged neutral fraction in our reionization model. Second, we consider stacking the transmission profile around highly absorbed regions in the forest. If and only if there is significantly neutral gas in the IGM, absorption in the damping wing of the Lyα line will cause the transmission to recover slowly as one moves from absorbed to transmitted portions of the spectrum. Third, the deuterium Lyβ line should imprint a small but distinctive absorption feature slightly blueward of absorbed neutral regions in the Lyβ forest. We show that these tests can be carried out with existing Keck HIRES spectra at z ∼ 5.5, with the damping wing being observable for 〈x{sub H} {sub I}〉≳0.05 and the deuterium feature observable with additional high-resolution spectra for 〈x{sub H} {sub I}〉≳0.2.

  4. Murine Visceral Leishmaniasis: IgM and Polyclonal B-Cell Activation Lead to Disease Exacerbation

    PubMed Central

    Deak, Eszter; Jayakumar, Asha; Wing Cho, Ka; Goldsmith-Pestana, Karen; Dondji, Blaise; Lambris, John D.; McMahon-Pratt, Diane

    2010-01-01

    In visceral leishmaniasis, the draining lymph node (DLN) is the initial site for colonization and establishment of infection after intradermal transmission by the sand fly vector; however, little is known about the developing immune response within this site. Using an intradermal infection model, which allows for parasite visceralization, we have examined the ongoing immune responses in the DLN of BALB/c mice infected with L. infantum. Although not unexpected, at early times post-infection there is a marked B cell expansion in the DLN, which persists throughout infection. However, the characteristics of this response were of interest; as early as day 7 post-infection, polyclonal antibodies (TNP, OVA, chromatin) were observed and the levels appeared comparable to the specific anti-leishmania response. Although B-cell-deficient JHD BALB/c mice are relatively resistant to infection, neither B-cell-derived IL-10 nor B-cell antigen presentation appear to be primarily responsible for the elevated parasitemia. However, passive transfer and reconstitution of JHD BALB/c with secretory immunoglobulins, (IgM or IgG; specific or non-specific immune complexes) results in increased susceptibility to L. infantum infection. Further, JHD BALB/c mice transgenetically reconstituted to secrete IgM demonstrated exacerbated disease in comparison to wild type BALB/c mice as early as 2 days post-infection. Evidence suggests that complement activation (generation of C5a) and signaling via the C5aR (CD88) is related to the disease exacerbation caused by IgM rather than cytokine levels (IL-10 or IFN-γ). Overall these studies indicate that polyclonal B cell activation, which is known to be associated with human visceral leishmaniasis, is an early and intrinsic characteristic of disease and may represent a target for therapeutic intervention. PMID:20213734

  5. A critical study of the use of staphylococci containing protein A for separation of IgG and IgM antibodies.

    PubMed

    Grangeot-Keros, L; Lebrun, L; Briantais, M J; Pillot, J

    1982-06-11

    This study was to determine the best conditions for using staphylococci bearing protein A to separate IgG from IgM. The validity of the technique was evaluated for detection of IgM with antimicrobial activity and for typing monoclonal IgM. The results indicate that separation of IgG and IgM is not entirely satisfactory in normal sera and worse in hyperglobulinemic sera. The detection and titration of IgM antimicrobial antibodies (rubella and hepatitis B core (HBc) specific IgM) was unreliable because IgG was only partially absorbed by staphylococcal cells, while a significant portion of IgM was bound. The use of higher concentrations of staphylococci did not improve the results because the more IgG was absorbed, the more IgM was also bound. It is shown that with anti-HBc specific IgM the risk of misinterpretation is very high with a sensitive radioimmunoassay technique allowing detection of trace amounts of nonabsorbed IgG. In contrast staphylococcal protein A proved useful in typing monoclonal IgM.

  6. Intrathecal somatic hypermutation of IgM in multiple sclerosis and neuroinflammation.

    PubMed

    Beltrán, Eduardo; Obermeier, Birgit; Moser, Markus; Coret, Francisco; Simó-Castelló, María; Boscá, Isabel; Pérez-Miralles, Francisco; Villar, Luisa M; Senel, Makbule; Tumani, Hayrettin; Hohlfeld, Reinhard; Casanova, Bonaventura; Dornmair, Klaus

    2014-10-01

    Intrathecal oligoclonal bands of the cerebrospinal fluid are considered the most important immunological biomarkers of multiple sclerosis. They typically consist of clonally expanded IgG antibodies that underwent affinity maturation during sustained stimulation by largely unknown antigens. In addition, ∼40% of patients with multiple sclerosis have oligoclonal bands that consist of expanded IgM antibodies. We investigated the molecular composition of IgM- and IgG-chains from cerebrospinal fluid of 12 patients with multiple sclerosis, seven patients with other neurological diseases, and eight healthy control subjects by high-throughput deep-sequencing and single-cell PCR. Further, we studied the expression of activation-induced cytidine deaminase, the key enzyme for affinity maturation of antibodies, in cerebrospinal fluid samples of 16 patients. From the cerebrospinal fluid of two multiple sclerosis patients we isolated single B cells and investigated the co-expression of antibody chains with activation-induced cytidine deaminase. In striking contrast to IgM-chains from peripheral blood, IgM-chains from cerebrospinal fluid of patients with multiple sclerosis or neuroborreliosis showed a high degree of somatic hypermutation. We found a high content of mutations that caused amino acid exchanges as compared to silent mutations. In addition, more mutations were found in the complementarity determining regions of the IgM-chains, which interact with yet unknown antigens, as compared to framework regions. Both observations provide evidence for antigen-driven affinity maturation. Furthermore, single B cells from the cerebrospinal fluid of patients with multiple sclerosis co-expressed somatically hypermutated IgM-chains and activation-induced cytidine deaminase, an enzyme that is crucial for somatic hypermutation and class switch recombination of antibodies and is normally expressed during activation of B cells in germinal centres. Clonal tracking of particular IgM(+) B

  7. Intrathecal somatic hypermutation of IgM in multiple sclerosis and neuroinflammation.

    PubMed

    Beltrán, Eduardo; Obermeier, Birgit; Moser, Markus; Coret, Francisco; Simó-Castelló, María; Boscá, Isabel; Pérez-Miralles, Francisco; Villar, Luisa M; Senel, Makbule; Tumani, Hayrettin; Hohlfeld, Reinhard; Casanova, Bonaventura; Dornmair, Klaus

    2014-10-01

    Intrathecal oligoclonal bands of the cerebrospinal fluid are considered the most important immunological biomarkers of multiple sclerosis. They typically consist of clonally expanded IgG antibodies that underwent affinity maturation during sustained stimulation by largely unknown antigens. In addition, ∼40% of patients with multiple sclerosis have oligoclonal bands that consist of expanded IgM antibodies. We investigated the molecular composition of IgM- and IgG-chains from cerebrospinal fluid of 12 patients with multiple sclerosis, seven patients with other neurological diseases, and eight healthy control subjects by high-throughput deep-sequencing and single-cell PCR. Further, we studied the expression of activation-induced cytidine deaminase, the key enzyme for affinity maturation of antibodies, in cerebrospinal fluid samples of 16 patients. From the cerebrospinal fluid of two multiple sclerosis patients we isolated single B cells and investigated the co-expression of antibody chains with activation-induced cytidine deaminase. In striking contrast to IgM-chains from peripheral blood, IgM-chains from cerebrospinal fluid of patients with multiple sclerosis or neuroborreliosis showed a high degree of somatic hypermutation. We found a high content of mutations that caused amino acid exchanges as compared to silent mutations. In addition, more mutations were found in the complementarity determining regions of the IgM-chains, which interact with yet unknown antigens, as compared to framework regions. Both observations provide evidence for antigen-driven affinity maturation. Furthermore, single B cells from the cerebrospinal fluid of patients with multiple sclerosis co-expressed somatically hypermutated IgM-chains and activation-induced cytidine deaminase, an enzyme that is crucial for somatic hypermutation and class switch recombination of antibodies and is normally expressed during activation of B cells in germinal centres. Clonal tracking of particular IgM(+) B

  8. Selective IgM deficiency in an adult presenting with Streptococcus pneumoniae septic arthritis.

    PubMed

    Phuphuakrat, Angsana; Ngamjanyaporn, Pintip; Nantiruj, Kanokrat; Luangwedchakarn, Voravich; Malathum, Kumthorn

    2016-02-01

    Septic arthritis caused by Streptococcus pneumoniae is uncommon. Most of the patients who have invasive pneumococcal infection have underlying diseases associated with impaired immune function. We report a case of polyarticular pneumococcal septic arthritis in a previously healthy adult as the first manifestation of selective immunoglobulin (Ig)M deficiency. The patient had no evidence of autoimmune disease or malignancy. Serum IgG, IgA, and complement levels were normal. Numbers of lymphocyte subsets were in normal range except that of CD4+ cells, which was slightly low. Invasive pneumococcal disease in a healthy adult should lead to further investigation for underlying diseases including primary immunodeficiencies.

  9. Complement deposition in autoimmune hemolytic anemia is a footprint for difficult-to-detect IgM autoantibodies.

    PubMed

    Meulenbroek, Elisabeth M; de Haas, Masja; Brouwer, Conny; Folman, Claudia; Zeerleder, Sacha S; Wouters, Diana

    2015-11-01

    In autoimmune hemolytic anemia autoantibodies against erythrocytes lead to increased clearance of the erythrocytes, which in turn results in a potentially fatal hemolytic anemia. Depending on whether IgG or IgM antibodies are involved, response to therapy is different. Proper identification of the isotype of the anti-erythrocyte autoantibodies is, therefore, crucial. However, detection of IgM autoantibodies can be challenging. We, therefore, set out to improve the detection of anti-erythrocyte IgM. Direct detection using a flow cytometry-based approach did not yield satisfactory improvements. Next, we analyzed whether the presence of complement C3 on a patient's erythrocytes could be used for indirect detection of anti-erythrocyte IgM. To this end, we fractionated patients' sera by size exclusion chromatography and tested which fractions yielded complement deposition on erythrocytes. Strikingly, we found that all patients with C3 on their erythrocytes according to standard diagnostic tests had an IgM anti-erythrocyte component that could activate complement, even if no such autoantibody had been detected with any other test. This also included all tested patients with only IgG and C3 on their erythrocytes, who would previously have been classified as having an IgG-only mediated autoimmune hemolytic anemia. Depleting patients' sera of either IgG or IgM and testing the remaining complement activation confirmed this result. In conclusion, complement activation in autoimmune hemolytic anemia is mostly IgM-mediated and the presence of covalent C3 on patients' erythrocytes can be taken as a footprint of the presence of anti-erythrocyte IgM. Based on this finding, we propose a diagnostic workflow that will aid in choosing the optimal treatment strategy.

  10. Complement deposition in autoimmune hemolytic anemia is a footprint for difficult-to-detect IgM autoantibodies

    PubMed Central

    Meulenbroek, Elisabeth M.; de Haas, Masja; Brouwer, Conny; Folman, Claudia; Zeerleder, Sacha S.; Wouters, Diana

    2015-01-01

    In autoimmune hemolytic anemia autoantibodies against erythrocytes lead to increased clearance of the erythrocytes, which in turn results in a potentially fatal hemolytic anemia. Depending on whether IgG or IgM antibodies are involved, response to therapy is different. Proper identification of the isotype of the anti-erythrocyte autoantibodies is, therefore, crucial. However, detection of IgM autoantibodies can be challenging. We, therefore, set out to improve the detection of anti-erythrocyte IgM. Direct detection using a flow cytometry-based approach did not yield satisfactory improvements. Next, we analyzed whether the presence of complement C3 on a patient’s erythrocytes could be used for indirect detection of anti-erythrocyte IgM. To this end, we fractionated patients’ sera by size exclusion chromatography and tested which fractions yielded complement deposition on erythrocytes. Strikingly, we found that all patients with C3 on their erythrocytes according to standard diagnostic tests had an IgM anti-erythrocyte component that could activate complement, even if no such autoantibody had been detected with any other test. This also included all tested patients with only IgG and C3 on their erythrocytes, who would previously have been classified as having an IgG-only mediated autoimmune hemolytic anemia. Depleting patients’ sera of either IgG or IgM and testing the remaining complement activation confirmed this result. In conclusion, complement activation in autoimmune hemolytic anemia is mostly IgM-mediated and the presence of covalent C3 on patients’ erythrocytes can be taken as a footprint of the presence of anti-erythrocyte IgM. Based on this finding, we propose a diagnostic workflow that will aid in choosing the optimal treatment strategy. PMID:26354757

  11. Comparison of a commercial dengue IgM capture ELISA with dengue antigen focus reduction microneutralization test and the Centers for Disease Control dengue IgM capture-ELISA.

    PubMed

    Welch, Ryan J; Chang, Gwong-Jen J; Litwin, Christine M

    2014-01-01

    Dengue (DENV) infection is caused by an arbovirus that is a member of the family Flaviviridae, genus Flavivirus. The diagnosis of acute dengue infection using clinical signs and symptoms can be difficult since the manifestations cannot be readily differentiated from other infections. Therefore the diagnosis of acute dengue infection depends upon laboratory assays. Dengue virus ELISAs have been designed for the detection of IgM and IgG antibodies in addition to nonstructural 1 (NS1) antigens. The InBios IgM Dengue ELISA was compared to the Antigen Focus Reduction Microneutralization Test (FRμNT90) and Centers for Disease Control Dengue IgM Capture-ELISA (CDC MAC-ELISA). The calculated sensitivity, specificity and agreement of the InBios ELISA compared to FRμNT90 and CDC MAC-ELISA was 88.7% (C.I. 81.4-93.7), 93.1% (C.I. 89.1-95.8) and 91.5% (C.I. 86.3-95.0), respectively. In summary the InBios IgM Dengue ELISA sensitivity and specificity is comparable to other commercially available IgM Capture-ELISAs.

  12. Generation, characterization and in vivo biological activity of two distinct monoclonal anti-PEG IgMs

    SciTech Connect

    Hashimoto, Yosuke; Shimizu, Taro; Mima, Yu; Abu Lila, Amr S.; Ishida, Tatsuhiro; Kiwada, Hiroshi

    2014-05-15

    PEGylation, the attachment of polyethylene glycol (PEG) to nanocarriers and proteins, is a widely accepted approach to improving the in vivo efficacy of the non-PEGylated products. However, both PEGylated liposomes and PEGylated proteins reportedly trigger the production of specific antibodies, mainly IgM, against the PEG moiety, which possibly leads to a reduction in safety and therapeutic efficacy of the PEGylated products. In the present study, two monoclonal anti-PEG IgMs — HIK-M09 via immunization with an intravenous injection of PEGylated liposomes (SLs) and HIK-M11 via immunization with a subcutaneous administration of PEGylated ovalbumin (PEG-OVA) were successfully generated. The generated IgMs showed efficient reactivity to mPEG{sub 2000} conjugated to 1,2-distearoyl-sn-glycero-3-phospho-ethanolamine (DSPE), PEGylated liposome (SL) and PEG-OVA. It appears that HIK-M09 recognizes ethoxy (OCH{sub 2}CH{sub 2}) repeat units along with a terminal motif of PEG, while HIK-M11 recognizes only ethoxy repeat units of PEG. Such unique properties allow HIK-M09 to bind with dense PEG. In addition, their impact on the in vivo clearance of the PEGylated products was investigated. It was found that the generated ant-PEG IgMs induced a clearance of SL as they were intravenously administered with SL. Interestingly, the HIK-M11, generated by PEG-OVA, induced the clearance of both SL and PEG-OVA, while the HIK-M09, generated by SL, induced the clearance of SL only. We here revealed that the presence of serum anti-PEG IgM and the subsequent binding of anti-PEG IgM to the PEGylated products are not necessarily related to the enhanced clearance of the products. It appears that subsequent complement activation following anti-PEG IgM binding is the most important step in dictating the in vivo fate of PEGylated products. This study may have implications for the design, development and clinical application of PEGylated products and therapeutics. - Highlights: • Two monoclonal

  13. Cold-induced precipitation of a monoclonal IgM: a negative activation enthalpy reaction.

    PubMed

    Meliga, Stefano C; Farrugia, William; Ramsland, Paul A; Falconer, Robert J

    2013-01-17

    Cold-induced precipitation of a monoclonal IgM cryoglobulin isolated from a patient with Waldenström's macroglobulinemia was observed to have a negative activation enthalpy. The rate of the reaction increased, as the temperature decreased. Differential scanning calorimetry of the monoclonal IgM showed precipitation as an inverted peak during a downward temperature scan. The transition temperature was between 14 and 15 °C and was possibly concentration dependent. At temperatures below the transition the precipitation was best described by second-order kinetics. The difference in change in enthalpy between precipitation and disassociation suggests that cold-induced precipitation had a fast precipitation stage followed by a slower consolidation reaction. Negligible curvature of the Eyring plot suggested the precipitation reaction was dominated by van der Waal forces and hydrogen bonding. Conversely, during an upward temperature scan, disassociation was observed as a positive enthalpy peak. This reaction had two stages, a reaction undoing consolidation followed by heat-induced disassociation that had first-order kinetics.

  14. Humoral immune response in patients with IgA and IgM glomerulonephritis.

    PubMed Central

    Pasternack, A; Mustonen, J; Leinikki, P

    1986-01-01

    A single dose of inactivated mumps virus vaccine was administered to male patients with IgA glomerulonephritis (IgA-GN), IgM glomerulonephritis (IgM-GN) and to healthy males. Antibodies to mumps virus were determined using an enzyme-linked immunosorbent assay. Patients with IgA-GN showed a higher and more sustained IgG and IgA antibody response compared to patients with IgM-GN or healthy controls. Before vaccination, patients with IgM-GN had higher levels of IgG antibodies than the controls or those with IgA-GN. However, the IgA antibody and IgG responses after vaccination were low. IgM antibody responses did not vary among the groups studied. It is concluded that patients with IgA-GN are high responders for IgA and IgG antibody production. Patients with IgM-GN are low responders, especially for IgA antibody. PMID:3955883

  15. Cold-induced precipitation of a monoclonal IgM: a negative activation enthalpy reaction.

    PubMed

    Meliga, Stefano C; Farrugia, William; Ramsland, Paul A; Falconer, Robert J

    2013-01-17

    Cold-induced precipitation of a monoclonal IgM cryoglobulin isolated from a patient with Waldenström's macroglobulinemia was observed to have a negative activation enthalpy. The rate of the reaction increased, as the temperature decreased. Differential scanning calorimetry of the monoclonal IgM showed precipitation as an inverted peak during a downward temperature scan. The transition temperature was between 14 and 15 °C and was possibly concentration dependent. At temperatures below the transition the precipitation was best described by second-order kinetics. The difference in change in enthalpy between precipitation and disassociation suggests that cold-induced precipitation had a fast precipitation stage followed by a slower consolidation reaction. Negligible curvature of the Eyring plot suggested the precipitation reaction was dominated by van der Waal forces and hydrogen bonding. Conversely, during an upward temperature scan, disassociation was observed as a positive enthalpy peak. This reaction had two stages, a reaction undoing consolidation followed by heat-induced disassociation that had first-order kinetics. PMID:23240622

  16. Recognition of a human T-lymphocyte differentiation antigen by an IgM monoclonal antibody.

    PubMed Central

    Bastin, J M; Granger, S; Tidman, N; Janossy, G; McMichael, A J

    1981-01-01

    A monoclonal antibody directed at a determinant on human T cells was produced and characterized. This IgM antibody, MBG6, bound to human peripheral blood T lymphocytes and to medullary thymocytes. It was unreactive with normal B cells, B-cell lines and granulocytes. Apart from T lymphocytes, bone marrow cells (including cells positive for the terminal transferase marker, myeloid colony-forming cells, myeloblasts, and differentiating myeloid and erythroid cells) were negative. Peripheral blood cells that were treated with MBG6 and rabbit complement were no longer capable of proliferating in response to phytohaemagglutinin or concanavalin A; MBG6 did not have any direct mitogenic action on T lymphocytes. Double immunofluorescence studies using IgM MBG6 and OKT3, and IgG2a monoclonal antibody that recognizes all peripheral T cells, showed that these two antibodies identified exactly the same cell populations. Competitive binding studies, however, indicated that MBG6 and OKT3 recognized different epitopes. The antibody may have clinical applications in bone marrow transplantation. PMID:6802543

  17. Assessment of snapper (Pagrus auratus) natural IgM binding to bromelain treated sheep erythrocytes.

    PubMed

    Morrison, Richard N; Bruce Lyons, A; Nowak, Barbara F; Hayball, John D

    2005-01-01

    Normal snapper (Pagrus auratus Bloch and Schneider) serum was examined for natural IgM that binds to protease (bromelain) treated sheep erythrocytes (BrSRBC) in a model assay system that has been used to appraise natural IgM of various mammals. Normal snapper serum lysed BrSRBC while haemolysis was abrogated by heat inactivation of serum and in divalent cation-deficient conditions, indicative of classical complement mediated lysis. In addition, heat inactivated normal snapper serum agglutinated BrSRBC while phosphatidylcholine (PtC) liposomes partially inhibited both haemolysis and agglutination. Inhibition of haemolysis and agglutination may have been mediated by an interaction between immunoglobulin (Ig) and PtC as protein A purified snapper Ig bound to PtC liposomes. However it is not known if this binding was PtC specific nor if the binding was initiated by either the Fab and/or Fc domains of snapper Ig. BrSRBC plaque forming cells (PFC) were detected in the peritoneal cavity, spleen, head kidney and peripheral blood of normal snapper. The greatest proportion of BrSRBC PFC per B cell was within the peritoneal cavity followed by the spleen, peripheral blood and head kidney. Together, these data suggest that normal snapper serum may contain natural Ig that binds BrSRBC, activating the classical complement cascade.

  18. Generation of IgM anti-platelet autoantibody in dengue patients.

    PubMed

    Lin, C F; Lei, H Y; Liu, C C; Liu, H S; Yeh, T M; Wang, S T; Yang, T I; Sheu, F C; Kuo, C F; Lin, Y S

    2001-02-01

    Dengue virus infection causes a wide range of diseases from dengue fever to life-threatening dengue hemorrhagic fever and dengue shock syndrome (DHF/DSS). The mechanisms involved in DHF/DSS pathogenesis remain unclear. Patient sera collected from an outbreak in southern Taiwan from November 1998 to January 1999 were studied. The presence of antibodies which cross-reacted with platelets could be detected in patient sera, and the isotype of these autoantibodies was IgM. The anti-platelet IgM levels were higher in DHF/DSS than in dengue fever patient sera in disease acute phase. These autoantibodies were still detectable in convalescent stage (1-3 weeks after acute phase) and even eight to nine months after illness. The platelet binding activity was not observed in other virus-infected patient sera tested. Further investigation showed that dengue patient sera caused platelet lysis in the presence of complement. The platelet cytotoxicity induced by DHF/DSS patient sera was higher than that by dengue fever sera. Dengue patient sera also inhibited platelet aggregation which, however, appeared to be not related to DHF/DSS development.

  19. [X-linked hyper-IGM syndrome associated to sclerosing cholangitis and gallbladder neoplasm: clinical case].

    PubMed

    Rodríguez, Cristián; Carrión, Flavio; Marinovic, María Angélica; Chávez, Eduardo; Preisler, Jessica; Pooley, Francisco; Futatani, Takeshi; Ochs, Hans D

    2003-03-01

    We report a 11 years old male diagnosed as a X-linked hyper-IgM syndrome that presented with recurrent infections and sclerosing cholangitis and later developed a gallbladder cancer. Immunological evaluation showed decreased levels of serum IgG and IgA with elevated levels of IgM. Study of CD40 ligand expression on mitogen activated peripheral blood mononuclear cells revealed total absence of this marker on T lymphocytes. Molecular analysis detected, in the patient and his mother, a nonsense mutation in exon 1 of the transmembrane segment of the CD40 ligand. He also presented elevation of alkaline phosphatases and mild elevation of liver enzymes. Liver biopsy demonstrated the presence of idiopathic sclerosing cholangitis. The patient was started on monthly IVIG therapy at 400 mg/kg, as well as ursodeoxycholic acid and vitamin E, with normalization of his IgG and IgM levels a decrease in the incidence of infections and normalization of liver function. Three years after diagnosis, we detected the presence of polyps inside the gallbladder that were reported at biopsy as adenocarcinoma. He underwent hepatic bisegmentectomy (VI B-V) and local lymphadenectomy.

  20. Gravity changes during animal development affect IgM heavy-chain transcription and probably lymphopoiesis.

    PubMed

    Huin-Schohn, Cécile; Guéguinou, Nathan; Schenten, Véronique; Bascove, Matthieu; Koch, Guillemette Gauquelin; Baatout, Sarah; Tschirhart, Eric; Frippiat, Jean-Pol

    2013-01-01

    Our previous research demonstrated that spaceflight conditions affect antibody production in response to an antigenic stimulation in adult amphibians. Here, we investigated whether antibody synthesis is affected when animal development occurs onboard a space station. To answer this question, embryos of the Iberian ribbed newt, Pleurodeles waltl, were sent to the International Space Station (ISS) before the initiation of immunoglobulin heavy-chain expression. Thus, antibody synthesis began in space. On landing, we determined the effects of spaceflight on P. waltl development and IgM heavy-chain transcription. Results were compared with those obtained using embryos that developed on Earth. We find that IgM heavy-chain transcription is doubled at landing and that spaceflight does not affect P. waltl development and does not induce inflammation. We also recreated the environmental modifications encountered by the embryos during their development onboard the ISS. This strategy allowed us to demonstrate that gravity change is the factor responsible for antibody heavy-chain transcription modifications that are associated with NF-κB mRNA level variations. Taken together, and given that the larvae were not immunized, these data suggest a modification of lymphopoiesis when gravity changes occur during ontogeny.

  1. Conserved natural IgM antibodies mediate innate and adaptive immunity against the opportunistic fungus Pneumocystis murina.

    PubMed

    Rapaka, Rekha R; Ricks, David M; Alcorn, John F; Chen, Kong; Khader, Shabaana A; Zheng, Mingquan; Plevy, Scott; Bengtén, Eva; Kolls, Jay K

    2010-12-20

    Host defense against opportunistic fungi requires coordination between innate and adaptive immunity for resolution of infection. Antibodies generated in mice vaccinated with the fungus Pneumocystis prevent growth of Pneumocystis organisms within the lungs, but the mechanisms whereby antibodies enhance antifungal host defense are poorly defined. Nearly all species of fungi contain the conserved carbohydrates β-glucan and chitin within their cell walls, which may be targets of innate and adaptive immunity. In this study, we show that natural IgM antibodies targeting these fungal cell wall carbohydrates are conserved across many species, including fish and mammals. Natural antibodies bind fungal organisms and enhance host defense against Pneumocystis in early stages of infection. IgM antibodies influence recognition of fungal antigen by dendritic cells, increasing their migration to draining pulmonary lymph nodes. IgM antibodies are required for adaptive T helper type 2 (Th2) and Th17 cell differentiation and guide B cell isotype class-switch recombination during host defense against Pneumocystis. These experiments suggest a novel role for the IgM isotype in shaping the earliest steps in recognition and clearance of this fungus. We outline a mechanism whereby serum IgM, containing ancient specificities against conserved fungal antigens, bridges innate and adaptive immunity against fungal organisms.

  2. High affinity IgM(+) memory B cells are generated through a germinal center-dependent pathway.

    PubMed

    Hara, Yasushi; Tashiro, Yasuyuki; Murakami, Akikazu; Nishimura, Miyuki; Shimizu, Takeyuki; Kubo, Masato; Burrows, Peter D; Azuma, Takachika

    2015-12-01

    During a T cell-dependent immune response, B cells undergo clonal expansion and selection and the induction of isotype switching and somatic hypermutation (SHM). Although somatically mutated IgM(+) memory B cells have been reported, it has not been established whether they are really high affinity B cells. We tracked (4-hydroxy-3-nitrophenyl) acetyl hapten-specific GC B cells from normal immunized mice based on affinity of their B cell receptor (BCR) and performed BCR sequence analysis. SHM was evident by day 7 postimmunization and increased with time, such that high affinity IgM(+) as well as IgG(+) memory B cells continued to be generated up to day 42. In contrast, class-switch recombination (CSR) was almost completed by day 7 and then the ratio of IgG1(+)/IgM(+) GC B cells remained unchanged. Together these findings suggest that IgM(+) B cells undergo SHM in the GC to generate high affinity IgM(+) memory cells and that this process continues even after CSR is accomplished.

  3. Binding of IgM rheumatoid factor to group A, C and G streptococci with IgG Fc receptors.

    PubMed

    Schröder, A K; Christensen, P; Svensson, M L

    1986-01-01

    The possibility that IgM rheumatoid factors bind to streptococci was studied. Using a sequence of Sephadex G200 gel filtration, protein A-Sepharose CL-4B chromatography and preparatory electrophoresis, IgM was isolated from the sera of 2 patients with rheumatoid arthritis and then radiolabelled with 125I. There was significant binding of radiolabelled IgM to group-A streptococci types M1, M15 and M22, and to a group-C and a group-G strain, all expressing IgG Fc receptors, but none to Staphylococcus aureus, Escherichia coli or to 11 other strains of streptococci without IgG Fc receptors. The radiolabelled IgM was separated by affinity chromatography on a column containing human IgG. Types M1 and M15 bound the fraction retained on the column, whereas M22 bound both this fraction and the non-retained fraction. Commercial human IgG, even at high concentrations, did not inhibit binding. The binding reaction, which is perhaps triggered either by the IgM rheumatoid factor or by IgG complexed with rheumatoid factor, could be a useful tool for removal of anti-immunoglobulin from the blood of patients with rheumatoid arthritis.

  4. IgG and IgM antibodies to viral glycoproteins in respiratory syncytial virus infections of graded severity.

    PubMed Central

    Toms, G L; Webb, M S; Milner, P D; Milner, A D; Routledge, E G; Scott, R; Stokes, G M; Swarbrick, A; Taylor, C E

    1989-01-01

    Serum antibodies to the fusion (F) and large glycoprotein (G) of respiratory syncytial virus in the serum of 57 infected infants were measured by enzyme linked immunosorbent assay (ELISA). Most serum samples taken at the time of admission to hospital contained antibodies to both glycoproteins, and overall there was no significant evidence of a selective deficiency of antibody to either viral antigen. Less than a quarter of the infants showed rising IgG antibody titres to either glycoprotein after infection, whereas over threequarters produced an IgM response. There was a significant correlation between IgG response to viral glycoproteins and the age of the infant. The correlation of age with the IgM response was less pronounced, and there was no correlation between serum IgG antibody derived transplancentally in the acute phase of infection and IgM response to either glycoprotein. Neither IgG or IgM responses correlated with a clinical assessment of the severity of infection in the infants. IgM responses, however, were weakly correlated with reduced secretion of infectious virus in the upper respiratory tract. PMID:2624472

  5. IgG, IgM, and IgA antinuclear antibodies in discoid and systemic lupus erythematosus patients.

    PubMed

    Jost, Sheridan A; Tseng, Lin-Chiang; Matthews, Loderick A; Vasquez, Rebecca; Zhang, Song; Yancey, Kim B; Chong, Benjamin F

    2014-01-01

    IgG antinuclear antibodies (ANAs) are elevated in patients with systemic lupus erythematosus (SLE) compared with patients with discoid lupus erythematosus (DLE). To provide an expanded immunologic view of circulating ANAs in lupus patients, we compared the expressions of IgG, IgM, and IgA ANAs in DLE and SLE patients. In this cross-sectional study, sera from age-, gender-, and ethnic-matched SLE (N = 35), DLE (N = 23), and normal patients (N = 22) were tested for IgG, IgM, and IgA ANAs using enzyme-linked immunosorbent assays (ELISAs) and indirect immunofluorescence (IIF) with monkey esophagus as substrate. ELISAs showed elevated levels of IgG ANA, IgM ANA, and IgG/IgM ANA ratios in SLE patients compared with DLE and normal patients. IgA ANA expression was higher in SLE and DLE patients versus normal patients. IIF studies showed higher percentages of patients positive for IgG, IgM, and IgA ANAs in the SLE group. Higher IgG/IgM ANA ratios in SLE than DLE show enhanced class-switching and a more sustained humoral response in SLE. They also suggest a potential connection of IgM ANAs with disease containment.

  6. Circulating microparticles carry oxidation-specific epitopes and are recognized by natural IgM antibodies1[S

    PubMed Central

    Tsiantoulas, Dimitrios; Perkmann, Thomas; Afonyushkin, Taras; Mangold, Andreas; Prohaska, Thomas A.; Papac-Milicevic, Nikolina; Millischer, Vincent; Bartel, Caroline; Hörkkö, Sohvi; Boulanger, Chantal M.; Tsimikas, Sotirios; Fischer, Michael B.; Witztum, Joseph L.; Lang, Irene M.; Binder, Christoph J.

    2015-01-01

    Oxidation-specific epitopes (OSEs) present on apoptotic cells and oxidized low density lipoprotein (OxLDL) represent danger-associated molecular patterns that are recognized by different arcs of innate immunity, including natural IgM antibodies. Here, we investigated whether circulating microparticles (MPs), which are small membrane vesicles released by apoptotic or activated cells, are physiological carriers of OSEs. OSEs on circulating MPs isolated from healthy donors and patients with ST-segment elevation myocardial infarction (STE-MI) were characterized by flow cytometry using a panel of OSE-specific monoclonal antibodies. We found that a subset of MPs carry OSEs on their surface, predominantly malondialdehyde (MDA) epitopes. Consistent with this, a majority of IgM antibodies bound on the surface of circulating MPs were found to have specificity for MDA-modified LDL. Moreover, we show that MPs can stimulate THP-1 (human acute monocytic leukemia cell line) and human primary monocytes to produce interleukin 8, which can be inhibited by a monoclonal IgM with specificity for MDA epitopes. Finally, we show that MDA+ MPs are elevated at the culprit lesion site of patients with STE-MI. Our results identify a subset of OSE+ MPs that are bound by OxLDL-specific IgM. These findings demonstrate a novel mechanism by which anti-OxLDL IgM antibodies could mediate protective functions in CVD. PMID:25525116

  7. The etiology of Rubella IgM positivity in patients with rubella-like illness in Iran from 2011 to 2013.

    PubMed

    Khorrami, Seyed Mahmood Seyed; Mokhtari-Azad, Talat; Yavarian, Jila; Nasab, Gazal Sadat Fatemi; Naseri, Maryam; Jandaghi, Nazanin Zahra Shafiei

    2015-11-01

    Rubella is a mild self-limiting contagious viral disease caused by the rubella virus (RV). Although symptoms are often mild, the concern is centralized around the possible effect on a fetus growth and development in case of primary infection during early months of pregnancy. Recently acquired rubella is commonly confirmed by RV-specific IgM antibody detection in the serum. However, rubella primary infection is not always the only cause of IgM positivity. Other possible causes of rubella IgM positivity may include IgM persistence following vaccination or naturally acquired infection or even re-infection. Moreover, nonspecific IgM reactivity can cause false-positive results. There are few articles to differentiate the aetiology of rash in rubella-like illnesses. However, limited studies have been conducted on clarifying the source of IgM positivity in these cases. This article reports the study of 10,896 clinical cases demonstrating rubella-like illness between 2011 and 2013 in Iran. The rate of IgM positivity among these cases was 0.52% (57 cases). As predicted based on the high coverage of vaccination in Iran fewer than 16% of cases with ELISA IgM positive result, were due to current rubella primary infections. The greater part of the positive IgM reactions occurred in cross reactivity with other viruses (31.6%) or in prolonged IgM response post vaccination (24.6%). This research confirmed that the positive result of rubella IgM assay in vaccinated individuals is mainly caused by prolonged IgM production, rubella re-infection, and false positivity due to infection with other viruses, rather than the rubella primary infection itself.

  8. 2D immunoblots show differential response of mouse IgG and IgM antibodies to antigens of mammary carcinoma 4 T1 cells

    PubMed Central

    2014-01-01

    Background Immunosuppression in breast cancer has been reported in women and in the highly metastatic mouse mammary tumor model 4 T1. The immunosuppressive environment complicates the use of the humoral response against the tumor as an immunodiagnostic tool. IgM has not been used in immunodiagnostic in part because its antitumor responses, both innate and adaptive, have not been studied in function of time in breast cancer. We show a new approach to analyzing the mouse humoral immune response, and compare the evolution with time of IgG and IgM responses against the antigens of 4 T1 cells. Methods The study is based on 2-dimensional immunoblotting detection of antigens from 4 T1 cells by the IgG and IgM antibodies in the serum of female mice injected with 4 T1 cells. Results There was a high variability in the intra-and inter-mouse response. Variability in the IgM response was manifested as a pattern of spots that could become a multibinomial variable of 0 and 1, which could represent a signature of the immune response. Different numbers of spots was found in the IgG and IgM responses from week 1 to 5. On average, the IgM had more but the IgG response decrease with the time. The natural IgM at t = 0 responds stronger than w1; the adaptive response of both IgM and IgG were elicited where, with the former being stronger better than the latter. Antigens that are recognized by some female mice in the first week are also recognized by other female mice at time 0. Contamination of the natural IgM makes difficult use the adaptive IgM as a tool for immunodiagnostic. Conclusions IgM and IgG response varied with the time and individuals. Spot variation in 2D pattern for the natural IgM could be expressed as a binomial signature, which opens up the way to correlate a particular pattern with resistance or susceptibility. This uncovers a battery of IgMs for each individual to confront cancer or infections. The possibility to differentiate between adaptive IgM antibodies

  9. IgM and IgG antibodies to hepatitis C virus in patients with mixed cryoglobulinaemia.

    PubMed Central

    L'Abbate, A; Cutrupi, S; Rognetta, M; Fabiano, C; Craxi, A

    1993-01-01

    To assess the relationship between hepatitis C virus (HCV) infection and essential mixed cryoglobulinaemia (EMC), sera from 23 patients with EMC were tested for IgG and IgM antibodies to HCV antigens and for HCV RNA. Quantitative HCV antibody studies were performed on serum and purified cryoglobulin fractions. HCV antibodies of both IgG and IgM class were found in 22 (96%) patients. Ten of these were also HCV-RNA positives. Higher titres of anti-HCV IgM were present in the 11 patients with evidence of liver damage. Anti-HCV IgG antibodies were shown to be concentrated in the IgG fraction of cryoglobulins in all eight patients studied. These results strongly suggest a role for HCV in the pathogenesis of EMC. PMID:7693384

  10. Specific recognition pattern of IgM and IgG antibodies produced in the course of experimental paracoccidioidomycosis.

    PubMed Central

    Vaz, C A; Mackenzie, D W; Hearn, V M; Camargo, Z P; Singer-Vermes, L M; Burger, E; Calich, V L

    1992-01-01

    Specific IgM and IgG responses to Paracoccidioides brasiliensis produced in resistant and susceptible mice during experimental paracoccidioidomycosis were examined by the immunoblotting procedure. Sera from infected mice recognized 51 antigen bands with apparent molecular masses from 8 to 86 kD. Sixteen of these were defined as major antigen bands because of almost universal presence of antibodies to them, and their intense staining. All sera, including those from normal control mice, tested for both IgM and IgG antibody reacted with the major E antigen which appeared as a large diffuse band from 43 to 47 kD. Comparisons between resistant and susceptible mice showed some significant differences in IgM responses to many antigen bands. While IgG responses were quite similar for both strains, differences were apparent in the response to the antigens at 62 and 68 kD. Images Fig. 1 Fig. 2 Fig. 3 PMID:1563097

  11. Serum IgG, IgM and slow alpha-globulin levels in carrageenan-treated rats.

    PubMed Central

    Fowler, E. F.; Thomson, A. W.

    1979-01-01

    Serum levels of IgM, IgG, slow alpha 1- and slow alpha 2-globulins were measured either by quantitative radial immunodiffusion (IgG) or immunoelectrophoresis (IgM and slow alpha-globulins) during the 3-week period after i.p. injection of 50 mg potassium carrageenan. There was a significant elevation in levels of IgM and slow alpha 1-globulin, maximal on Day 4 and returning to normal by Day 14. Slow alpha 2-globulin was detectable within 24 h, reached a peak at Day 2, and was no longer measurable in most rats by Day 14. Levels of IgG however, were unaffected by carrageenan injection. PMID:92333

  12. Severe autoimmune hemolytic anemia associated with IgM warm auto-antibodies in primary Sjögren's syndrome.

    PubMed

    Shinoda, Koichiro; Taki, Hirofumi; Hounoki, Hiroyuki; Ogawa, Reina; Sugiyama, Eiji; Tobe, Kazuyuki

    2010-02-01

    Primary Sjögren's syndrome is an autoimmune disorder involving mainly salivary and lachrymal glands. However, many extraglandular symptoms have also been reported. Although leucocytopenia and lymphocytopenia are frequently observed in hematological disorders, autoimmune hemolytic anemia is rarely reported. We experienced a case of primary Sjögren's syndrome developing severe autoimmune hemolytic anemia. The patient's red blood cells showed spontaneous agglutination in saline at room temperature, and immunoglobulin M (IgM) was detected on the surface of red blood cells by flow cytometry, indicating that autoimmune hemolytic anemia was caused by warm reactive IgM antibodies. Immediate corticosteroid therapy resulted in a dramatic recovery. We report a first case of severe autoimmune hemolytic anemia caused by warm reactive IgM antibodies in primary Sjögren's syndrome.

  13. Functional affinity of IgM rheumatoid factor in patients with rheumatoid arthritis and other autoimmune diseases.

    PubMed Central

    Rath, S; Hogben, D N; Devey, M E

    1988-01-01

    The functional affinity of IgM rheumatoid factors (RF) was measured in 31 patients with rheumatoid arthritis (RA), 24 with systemic lupus erythematosus (SLE), 13 with Sjögren's syndrome (SS), and in 13 seropositive healthy individuals. The functional affinity of IgM RF from patients with RA was significantly lower than in the other clinical groups studied. In addition, there was a significant inverse correlation between functional affinity and titre of IgM RF in all the groups. These results suggest that the usual mechanisms of affinity based selective pressure (somatic diversification and antigen selection) may operate differently for autoantibodies to serum antigens such as IgG. PMID:3365027

  14. The Fifth Domain of Beta 2 Glycoprotein I Protects from Natural IgM Mediated Cardiac Ischaemia Reperfusion Injury

    PubMed Central

    Beretov, Julia; Atsumi, Tatsuya; Qi, Miao; Bhindi, Ravinay; Qi, Jian C.; Madigan, Michele C.

    2016-01-01

    Reperfusion after a period of ischemia results in reperfusion injury (IRI) which involves activation of the inflammatory cascade. In cardiac IRI, IgM natural antibodies (NAb) play a prominent role through binding to altered neoepitopes expressed on damaged cells. Beta 2 Glycoprotein I (β2GPI) is a plasma protein that binds to neoepitopes on damaged cells including anionic phospholipids through its highly conserved Domain V. Domain I of β2GPI binds circulating IgM NAbs and may provide a link between the innate immune system, IgM NAb binding and cardiac IRI. This study was undertaken to investigate the role of Β2GPI and its Domain V in cardiac IRI using wild-type (WT), Rag-1 -/- and β2GPI deficient mice. Compared with control, treatment with Domain V prior to cardiac IRI prevented binding of endogenous β2GPI to post-ischemic myocardium and resulted in smaller myocardial infarction size in both WT and β2GPI deficient mice. Domain V treatment in WT mice also resulted in less neutrophil infiltration, less apoptosis and improved ejection fraction at 24 h. Rag-1 -/- antibody deficient mice reconstituted with IgM NAbs confirmed that Domain V prevented IgM NAb induced cardiac IRI. Domain V remained equally effective when delivered at the time of reperfusion which has therapeutic clinical relevance.Based upon this study Domain V may function as a universal inhibitor of IgM NAb binding in the setting of cardiac IRI, which offers promise as a new therapeutic strategy in the treatment of cardiac IRI. PMID:27031114

  15. Generation of a novel high-affinity monoclonal antibody with conformational recognition epitope on human IgM.

    PubMed

    Sarikhani, Sina; Mirshahi, Manouchehr; Gharaati, Mohammad Reza; Mirshahi, Tooran

    2010-11-01

    As IgM is the first isotype of antibody which appears in blood after initial exposure to a foreign antigen in the pattern of primary response, detection, and quantification of this molecule in blood seems invaluable. To approach these goals, generation, and characterization of a highly specific mAb (monoclonal antibody) against human IgM were investigated. Human IgM immunoglobulins were used to immunize Balb/c mice. Spleen cells taken from the immunized animals were fused with SP2/O myeloma cells using PEG (polyethylene glycol, MW 1450) as fusogen. The hybridomas were cultured in HAT containing medium and supernatants from the growing hybrids were screened by enzyme-linked immunosorbent assay (ELISA) using plates coated with pure human IgM and the positive wells were then cloned at limiting dilutions. The best clone designated as MAN-1, was injected intraperitoneally to some Pristane-injected mice. Anti-IgM mAb was purified from the animals' ascitic fluid by protein-G sepharose followed by DEAE-cellulose ion exchange chromatography. MAN-1 interacted with human IgM with a very high specificity and affinity. The purity of the sample was tested by SDS-PAGE and the affinity constant was measured (K(a) = 3.5 x 10(9)M(-1). Immunoblotting and competitive ELISA were done and the results showed that the harvested antibody recognizes a conformational epitope on the mu chain of human IgM and there was no cross-reactivity with other subclasses of immunoglobulins. Furthermore, isotyping test was done and the results showed the subclass of the obtained mAb which was IgG(1)kappa. PMID:20162378

  16. Serum anti-collagen type IV IgM antibodies and development of diabetic nephropathy in diabetics with essential hypertension

    PubMed Central

    Tsinlikov, Ivan; Tsinlikova, Ivanka; Nicoloff, George; Blazhev, Alexander; Garev, Antoan

    2016-01-01

    Introduction and aims Arterial hypertension and diabetic vascular complications are connected with an elevated degradation of elastic tissue. This process leads to an increased production of antibodies to collagen type IV (ACIV Abs). In the present investigation we studied whether the serum levels of antibodies (IgG, IgM and IgA) to collagen are related with microvascular complications. Material and methods Serum levels of antibodies to collagen type IV (ACIV) IgG, IgM and IgA were measured using an ELISA method in 93 patients with type 2 diabetes mellitus and arterial hypertension (AH) (mean age 61.4 ±11.3 years, diabetes duration 9.88 ±3.12 years; hypertension duration 9.28 ±4.98). These values were compared to serum antibodies to CIV in 42 age and sex matched controls. Results ACIV IgM antibodies levels in patients with AH and T2DM were statisticaly significantly higher than controls 0.178 (0.145÷0.220) vs. 0.142 (0.118÷0.173) (KW = 6.31; p = 0.01). Group 1 (patients with microvascular complications) showed significantly higher levels of ACIV IgM than controls 0.180 (0.136÷0.223) vs. 0.142 (0.118÷0.173) (KW = 5.03; p = 0.02). Patients from Group 2 showed statistically significantly higher levels of ACIV IgM than controls 0.176 (0.151÷0.202) vs. 0.142 (0.118÷0.173) (KW = 6.15; p = 0.01). ACIV IgM antibodies showed correlation with microalbuminuria (r = 0.21); (p = 0.04), BMI (r = 0.19); (p = 0.04), creatinine clearance (r = –0.36); (p = 0.01) and GFR (r = –0.34); (p = 0.02). Conclusions Our study showed an association between elevation of serum levels of ACIV IgM and development of diabetic nephropathy. We suggest that levels of ACIV IgM can be useful method for identfying a high risk for development of diabetic nephropathy. PMID:27095927

  17. Complementary innate (anti-A-specific) IgM emerging from ontogenic O-GalNAc-transferase depletion: (Innate IgM complementarity residing in ancestral antigen completeness).

    PubMed

    Arend, Peter

    2014-04-01

    The murine and the human genome have global properties in common. So the murine anti-A-specific complementary IgM and related human innate isoagglutinin represent developmental, 2-mercaptoethanol-sensitive, complement-binding glycoproteins, which do not arise from any measurable environmentally-induced or auto- immune response. The murine anti-A certainly originates from a cell surface- or cell adhesion molecule, which in the course of germ cell development becomes devoid of O-GalNAc-transferase and is released into the circulation. In human sera the enzyme occurs exclusively in those of blood group A- and AB subjects, while in group O(H) an identically encoded protein lets expect an opposite function and appears in conjunction with a complementary anti-A reactive glycoprotein. Since O-glycosylations rule the carbohydrate metabolism in growth and reproduction processes, we propose that the ancestral histo-(blood)-group A molecule arises in the course of O-GalNAc-glycosylations of glycolipids and protein envelops at progenitor cell surfaces. Germ cell development postulates embryonic stem cell fidelity, which is characterised by persistent production of α-linked O-GalNAc-glycans. They are determined by the A-allele within the human, "complete" histo (blood) group AB(O) structure that in early ontogeny is hypothesised to be synthesised independently from the final phenotype. The structure either passes "completely" through the germline, in transferase-secreting mature tissues becoming the "complete" phenotype AB, or disappears in exhaustive glycotransferase depletion from the differentiating cell surfaces and leaves behind the "incomplete" blood group O-phenotype, which has released a transferase- and O-glycan-depleted, complementary glycoprotein (IgM) into the circulation. The process implies, that in humans the different blood phenotypes evolve from a "complete" AB(O) molecular complex in a distinct enzymatic and/or complement cascade suggesting O

  18. Exploring the relationship between anti-PEG IgM behaviors and PEGylated nanoparticles and its significance for accelerated blood clearance.

    PubMed

    Shiraishi, Kouichi; Kawano, Kumi; Maitani, Yoshie; Aoshi, Taiki; Ishii, Ken J; Sanada, Yusuke; Mochizuki, Shinichi; Sakurai, Kazuo; Yokoyama, Masayuki

    2016-07-28

    Surface PEGylation on nanoparticles has greatly helped prolong their blood circulation half-lives. However, The injection of PEGylated nanoparticles into mice induced poly(ethylene glycol) (PEG)-specific IgM antibodies (anti-PEG IgMs), significantly changing PEG-liposomes' pharmacokinetics. In this study, we used various PEG-conjugates to conduct a mechanistic study of anti-PEG IgMs' binding behavior. The conventional belief has been that anti-PEG IgMs bind to PEG main chains; however, our findings reveal that anti-PEG IgMs did not bind to PEG main chains, whereas anti-PEG IgMs did bind to PEG-hydrophobic polymer blocks. The insertion of a hydrophilic polymer between each PEG chain and each hydrophobic polymer block suppressed anti-PEG IgMs' binding. We prove here that hydrophobic blocks are essential to anti-PEG IgMs' binding, and also that anti-PEG IgMs do not bind to intact PEGs without hydrophobic moiety. These results support our conclusion that anti-PEG IgMs exhibit specificity to PEG; however, the presence of a hydrophobic block at a proximity position from each PEG chain is essential for the binding. Also in the present study, we elucidate relations between anti-PEG IgMs and PEGylated nanoparticles. In one of our previous studies, anti-PEG IgMs scarcely affected the pharmacokinetics of PEG-b-poly(β-benzyl l-aspartate) block copolymer (PEG-PBLA) micelles, whereas anti-PEG IgMs significantly decreased PEG-liposomes' blood circulation half-life. Finally, we found that the ratio of anti-PEG IgM molecules to PEG-liposome particles is critical to these pharmacokinetic changes, and that a 10-fold increase in the number of anti-PEG IgM molecules permitted them to capture the PEG-liposome particles, thus leading to the aforementioned changes. PMID:27164541

  19. Rapid immunoenzymatic technique for titration of rabies antibodies IgG and IgM results.

    PubMed

    Atanasiu, P; Savy, V; Gibert, C

    1978-11-17

    Techniques usually employed for the detection of rabies' antibodies are costly, time consuming, and sometimes fail to detect early antibodies. The introduction of immunoenzymatic techniques in the serology of viral disease represents a new and important advance. We therefore adapted this technique to the detection of rabies antibodies. We have found that the sera from rabies patients who had not received antirabies treatment do not show seroneutralizing antibodies until several days after the onset of symptoms. However, antibodies can be detected some days earlier by the immunoenzymatic method in the same samples. Furthermore, the immunoenzymatic test was applied to the detection of both the IgM or the IgG class of antirabies antibodies using an antihuman Ig-or antihuman IgG-peroxydase conjugate.

  20. Comparison of five different methods of rubella IgM antibody testing.

    PubMed Central

    Cubie, H; Edmond, E

    1985-01-01

    Five tests for the detection of rubella specific IgM antibody were compared. They were the conventional method of sucrose density gradient fractionation, followed by haemagglutination inhibition; an anti-mu capture radioimmunoassay; and three commercially available enzyme linked assays: Rubazyme M, Rubenz M I, and its successor, Rubenz M II. The five methods detected similar numbers of rubella positive samples between seven and 35 days after the onset of symptoms; in the earlier stages, however, the radioimmunoassay and Rubenz M II were more sensitive. All three commercial kits were straightforward to use but produced misleading positive results with sera containing heterophil antibody. In considering sensitivity, specificity, and cost effectiveness together the Rubenz M tests were the most appropriate for routine use. With the recent withdrawal of Rubenz M I from the market only Rubenz M II is now available. If Epstein-Barr virus infection is excluded, Rubenz M II provides a reliable test for the diagnostic laboratory. PMID:3968218

  1. Call admission control for CDMA systems with Interference Guard Margin (IGM)

    NASA Astrophysics Data System (ADS)

    Chen, Huan; Kumar, Sunil; Kuo, C.-C. Jay

    2002-01-01

    A call admission control (CAC) scheme and a resource-reservation estimation (RRE) method suitable for the interference-based wireless system, such as wide-band code division multiple access (W-CDMA), are proposed in this work. The proposed CAC scheme gives preferential treatment to high priority handoff calls by pre-reserving a certain amount of interference margin called the interference guard margin (IGM). The amount of guard margin is determined by the measurement performed by the RRE module in base stations. Each RRE module dynamically adjusts the level of guard margin by considering traffic conditions in neighboring cells based upon handoff requests. A service model is adopted to support multiple services, which includes mobile terminal's data rate, different levels of priorities, mobility and rate adaptivity characteristics. Simulations are conducted with OPNET to study the performance of the proposed scheme in terms of the objective function, blocking probabilities and system utilization under different traffic conditions.

  2. Reduced secreted mu mRNA synthesis in selective IgM deficiency of Bloom's syndrome.

    PubMed Central

    Kondo, N; Ozawa, T; Kato, Y; Motoyoshi, F; Kasahara, K; Kameyama, T; Orii, T

    1992-01-01

    Serum IgM concentrations were low although serum IgG and IgA concentrations were normal in both our patients with Bloom's syndrome. Although the percentages of surface IgM-bearing cells were not reduced, the numbers of IgM-secreting cells were markedly reduced. The membrane-bound mu (microns) and secreted mu (microseconds) mRNAs are produced from transcripts of a single immunoglobulin mu gene by alternative RNA processing pathways. The control of microseconds mRNA synthesis depends on the addition of poly(A) to microseconds C-terminal segment. In both patients, mu mRNA was well detected but microseconds C-terminal mRNA was scarcely detected, suggesting that microns mRNA was well transcribed but microseconds mRNA was not. There was, at least, no mutation or deletion in the microseconds C-terminal coding sequence, the RNA splice site (GG/TAAAC) at the 5' end of microseconds C-terminal segment and the AATAAA poly(A) signal sequence in both patients. Our results suggest that selective IgM deficiency in Bloom's syndrome is due to an abnormality in the maturation of surface IgM-bearing B cells into IgM-secreting cells and a failure of microseconds mRNA synthesis. Moreover, reduced microseconds mRNA synthesis may be due to the defect on developmental regulation of the site at which poly(A) is added to transcripts of the mu gene. Images Fig. 2 PMID:1563106

  3. Identification of IgM as a contaminant in lectin-FLISA assays for HCC detection.

    PubMed

    Wang, Mengjun; Comunale, Mary Ann; Herrera, Harmin; Betesh, Lucy; Kono, Yuko; Mehta, Anand

    2016-07-29

    Liver disease, in the form of hepatocellular carcinoma (HCC) accounts for > 700,000 deaths worldwide. A major reason for this is late diagnosis of HCC. The currently used biomarker, serum alpha-fetoprotein (AFP) is elevated in 40-60% of those with HCC and other markers that can either compliment or replace AFP are desired. Our previous work has identified a number of proteins that contain altered glycans in HCC. Specifically, these altered glycans were increased levels of core and outer arm fucosylation. To determine the clinical usefulness of those identified glycoproteins, a plate based assay was developed that allowed for the detection of fucosylated glycoforms. While this method was applicable to a number of independent patient sets, it was unable to specifically detect fucosylated glycoforms in many patient samples. That is, some material was present in serum that led to non-specific signal in the lectin- fluorescence -linked immunosorbent assay (lectin-FLISA). To address this issue, a systematic process was undertaken to identify the material. This material was found to be increased levels of lectin reactive IgM. Removal of both IgG and IgM using a multi-step protein A/G incubation and filtration step removed the contaminating signal and allowed for the analysis of specific protein glycoforms. This assay was subsequently used on two sample sets, one that was shown previously to be unable to be tested via a lectin FLISA and in a larger independent sample set. The clinical usefulness of this assay in the early detection of HCC is discussed.

  4. Identification of IgM as a contaminant in lectin-FLISA assays for HCC detection.

    PubMed

    Wang, Mengjun; Comunale, Mary Ann; Herrera, Harmin; Betesh, Lucy; Kono, Yuko; Mehta, Anand

    2016-07-29

    Liver disease, in the form of hepatocellular carcinoma (HCC) accounts for > 700,000 deaths worldwide. A major reason for this is late diagnosis of HCC. The currently used biomarker, serum alpha-fetoprotein (AFP) is elevated in 40-60% of those with HCC and other markers that can either compliment or replace AFP are desired. Our previous work has identified a number of proteins that contain altered glycans in HCC. Specifically, these altered glycans were increased levels of core and outer arm fucosylation. To determine the clinical usefulness of those identified glycoproteins, a plate based assay was developed that allowed for the detection of fucosylated glycoforms. While this method was applicable to a number of independent patient sets, it was unable to specifically detect fucosylated glycoforms in many patient samples. That is, some material was present in serum that led to non-specific signal in the lectin- fluorescence -linked immunosorbent assay (lectin-FLISA). To address this issue, a systematic process was undertaken to identify the material. This material was found to be increased levels of lectin reactive IgM. Removal of both IgG and IgM using a multi-step protein A/G incubation and filtration step removed the contaminating signal and allowed for the analysis of specific protein glycoforms. This assay was subsequently used on two sample sets, one that was shown previously to be unable to be tested via a lectin FLISA and in a larger independent sample set. The clinical usefulness of this assay in the early detection of HCC is discussed. PMID:27181357

  5. Production and characterization of monoclonal antibodies to IgM of Pacific herring (Clupea pallasii).

    PubMed

    Purcell, Maureen K; Bromage, Erin S; Silva, Jessica; Hansen, John D; Badil, Samantha M; Woodson, James C; Hershberger, Paul K

    2012-09-01

    Pacific herring (Clupea pallasii) have a central role in the North Pacific ecosystem as a forage fish species and are natural reservoirs of several important finfish pathogens, including Viral hemorrhagic septicemia virus (VHSV). Here, we report the identification of the gene encoding the immunoglobulin mu (IgM) heavy chain, as well as the development and characterization of monoclonal antibodies (MAbs) that specifically react with Pacific herring IgM. Pacific herring immunoglobulin was purified and consisted of heavy and light chains of approximately 80 and 25 kDa. Three hybridoma clones were initially identified by ELISA as reactive with purified immunoglobulin but only one clone was able to detect an 80 kDa protein in Pacific and Atlantic herring (Clupea harengus) whole plasma by denaturing western blot. However, all three MAbs were able to precipitate an 80 kDa protein from Pacific herring and LCMS sequencing of peptide fragments derived from this protein matched the predicted amino acid sequence of the cloned, heavy chain gene. In addition, two of the MAbs stained cells within the putative lymphocyte gates for the spleen, anterior kidney and posterior kidney but were not reactive for myeloid/granulocyte gates, which is consistent with these MAbs reacting with surface IgM⁺ B-cells. To our knowledge, this is the first report of IgM-related gene sequences and anti-IgM monoclonal antibodies from any member of the family Clupeidae. The antibodies produced in this study are critical for achieving our long-term goal of conducting serological surveillance to assess pathogen exposure in natural populations of Pacific herring.

  6. Complement activation by apoptotic cells occurs predominantly via IgM and is limited to late apoptotic (secondary necrotic) cells.

    PubMed

    Zwart, Bas; Ciurana, Caroline; Rensink, Irma; Manoe, Rishi; Hack, C Erik; Aarden, Lucien A

    2004-03-01

    Apoptotic cells activate complement via various molecular mechanisms. It is not known which of these mechanisms predominate in a physiological environment. Using Jurkat cells as a model, we investigated complement deposition on vital, early and late apoptotic (secondary necrotic) cells in a physiological medium, human plasma, and established the main molecular mechanism involved in this activation. Upon incubation with recalcified plasma, binding of C3 and C4 to early apoptotic cells was similar to background binding on vital cells. In contrast, late apoptotic (secondary necrotic) cells consistently displayed substantial binding of C4 and C3 and low, but detectable, binding of C1q. Binding of C3 and C4 to the apoptotic cells was abolished by EDTA or Mg-EGTA, and also by C1-inhibitor or a monoclonal antibody that inhibits C1q binding, indicating that complement fixation by the apoptotic cells was mainly dependent on the classical pathway. Late apoptotic cells also consistently bound IgM, in which binding significantly correlated with that of C4 and C3. Depletion of plasma for IgM abolished most of the complement fixation by apoptotic cells, which was restored by supplementation with purified IgM. We conclude that complement binding by apoptotic cells in normal human plasma occurs mainly to late apoptotic, secondary necrotic cells, and that the dominant mechanism involves classical pathway activation by IgM.

  7. Introduction of germline residues improves the stability of anti-HIV mAb 2G12-IgM

    PubMed Central

    Chromikova, Veronika; Mader, Alexander; Hofbauer, Stefan; Göbl, Christoph; Madl, Tobias; Gach, Johannes S.; Bauernfried, Stefan; Furtmüller, Paul G.; Forthal, Donald N.; Mach, Lukas; Obinger, Christian; Kunert, Renate

    2015-01-01

    Immunoglobulins M (IgMs) are gaining increasing attention as biopharmaceuticals since their multivalent mode of binding can give rise to high avidity. Furthermore, IgMs are potent activators of the complement system. However, they are frequently difficult to express recombinantly and can suffer from low conformational stability. Here, the broadly neutralizing anti-HIV-1 antibody 2G12 was class-switched to IgM and then further engineered by introduction of 17 germline residues. The impact of these changes on the structure and conformational stability of the antibody was then assessed using a range of biophysical techniques. We also investigated the effects of the class switch and germline substitutions on the ligand-binding properties of 2G12 and its capacity for HIV-1 neutralization. Our results demonstrate that the introduced germline residues improve the conformational and thermal stability of 2G12-IgM without altering its overall shape and ligand-binding properties. Interestingly, the engineered protein was found to exhibit much lower neutralization potency than its wild-type counterpart, indicating that potent antigen recognition is not solely responsible for IgM-mediated HIV-1 inactivation. PMID:25748881

  8. B-1 Cell Heterogeneity and the Regulation of Natural and Antigen-Induced IgM Production

    PubMed Central

    Baumgarth, Nicole

    2016-01-01

    A small subset of B cells, termed B-1 cells, with developmental origins, phenotypes, and functions that are distinct from those of conventional B cells exist in mice. It contributes the vast majority of spontaneously produced “natural” IgM. Natural IgM is constitutively produced, even in the absence of microbiota, and fulfills many distinct functions in tissue homeostasis and host defense. B-1 cells also respond with IgM production to innate signals and pathogen exposure, while maintaining steady-state levels natural IgM. Thus, within the B-1 cell pool, cells of distinct and heterogeneous functionality must exist to facilitate these different functions. This review considers three factors that may contribute to this heterogeneity: first, developmental differences regarding the origins of the precursors, second, tissue-specific signals that may differentially affect B-1 cells in the tissue compartments, and finally responsiveness to self-antigens as well as innate and antigen-specific signals. All three are likely to shape the repertoire and responsiveness of B-1 cells to homeostatic- and antigen-induced signals and thus contribute to the functional heterogeneity among these innate-like B cells. PMID:27667991

  9. The murine IgM secretory poly(A) site contains dual upstream and downstream elements which affect polyadenylation.

    PubMed Central

    Phillips, C; Virtanen, A

    1997-01-01

    Regulation of polyadenylation efficiency at the secretory poly(A) site plays an essential role in gene expression at the immunoglobulin (IgM) locus. At this poly(A) site the consensus AAUAAA hexanucleotide sequence is embedded in an extended AU-rich region and there are two downstream GU-rich regions which are suboptimally placed. As these sequences are involved in formation of the polyadenylation pre-initiation complex, we examined their function in vivo and in vitro . We show that the upstream AU-rich region can function in the absence of the consensus hexanucleotide sequence both in vivo and in vitro and that both GU-rich regions are necessary for full polyadenylation activity in vivo and for formation of polyadenylation-specific complexes in vitro . Sequence comparisons reveal that: (i) the dual structure is distinct for the IgM secretory poly(A) site compared with other immunoglobulin isotype secretory poly(A) sites; (ii) the presence of an AU-rich region close to the consensus hexanucleotide is evolutionarily conserved for IgM secretory poly(A) sites. We propose that the dual structure of the IgM secretory poly(A) site provides a flexibility to accommodate changes in polyadenylation complex components during regulation of polyadenylation efficiency. PMID:9171084

  10. Isolation and Characterization of IgM and IgY Antibodies from Plasma of Magellanic Penguins (Spheniscus magellanicus).

    PubMed

    Bizelli, Camila C; Silva, A Sandriana R; da Costa, Jessica D; Vanstreels, Ralph E T; Atzingen, Marina V; Santoro, Marcelo L; Fernandes, Irene; Catão-Dias, José L; Faquim-Mauro, Eliana L

    2015-03-01

    Infectious diseases such as aspergillosis, avian malaria, and viral infections are significant threats to the conservation of penguins, leading to morbidity and mortality of these birds both in captivity and in the wild. The immune response to such infectious diseases is dependent on different mechanisms mediated by cells and soluble components such as antibodies. Antibodies or immunoglobulins are glycoproteins that have many structural and functional features that mediate distinct effector immune functions. Three distinct classes of antibodies have been identified in birds: immunoglobulin A (IgA), immunoglobulin M (IgM), and immunoglobulin Y (IgY). In this study we aim to establish an efficient laboratory method to obtain IgM and IgY antibodies from plasma samples of healthy adult Magellanic penguins (Spheniscus magellanicus). The protocol was developed combining plasma delipidation, sequential precipitation with caprylic acid and ammonium sulfate, and size-exclusion chromatography. The efficiency of the protocol and the identity of the purified IgM and IgY antibodies were confirmed through enzyme-linked immunosorbent assay, Western blotting, one-dimensional and two-dimensional polyacrylamide gel electrophoresis, and lectin binding assay. Structural and physicochemical properties of IgM and IgY from Magellanic penguins were consistent with those of other avian species. This purification protocol will allow for more detailed studies on the humoral immunity of penguins and for the development of high specificity serologic assays to test Magellanic penguins for infectious pathogens.

  11. B-1 Cell Heterogeneity and the Regulation of Natural and Antigen-Induced IgM Production

    PubMed Central

    Baumgarth, Nicole

    2016-01-01

    A small subset of B cells, termed B-1 cells, with developmental origins, phenotypes, and functions that are distinct from those of conventional B cells exist in mice. It contributes the vast majority of spontaneously produced “natural” IgM. Natural IgM is constitutively produced, even in the absence of microbiota, and fulfills many distinct functions in tissue homeostasis and host defense. B-1 cells also respond with IgM production to innate signals and pathogen exposure, while maintaining steady-state levels natural IgM. Thus, within the B-1 cell pool, cells of distinct and heterogeneous functionality must exist to facilitate these different functions. This review considers three factors that may contribute to this heterogeneity: first, developmental differences regarding the origins of the precursors, second, tissue-specific signals that may differentially affect B-1 cells in the tissue compartments, and finally responsiveness to self-antigens as well as innate and antigen-specific signals. All three are likely to shape the repertoire and responsiveness of B-1 cells to homeostatic- and antigen-induced signals and thus contribute to the functional heterogeneity among these innate-like B cells.

  12. B-1 Cell Heterogeneity and the Regulation of Natural and Antigen-Induced IgM Production.

    PubMed

    Baumgarth, Nicole

    2016-01-01

    A small subset of B cells, termed B-1 cells, with developmental origins, phenotypes, and functions that are distinct from those of conventional B cells exist in mice. It contributes the vast majority of spontaneously produced "natural" IgM. Natural IgM is constitutively produced, even in the absence of microbiota, and fulfills many distinct functions in tissue homeostasis and host defense. B-1 cells also respond with IgM production to innate signals and pathogen exposure, while maintaining steady-state levels natural IgM. Thus, within the B-1 cell pool, cells of distinct and heterogeneous functionality must exist to facilitate these different functions. This review considers three factors that may contribute to this heterogeneity: first, developmental differences regarding the origins of the precursors, second, tissue-specific signals that may differentially affect B-1 cells in the tissue compartments, and finally responsiveness to self-antigens as well as innate and antigen-specific signals. All three are likely to shape the repertoire and responsiveness of B-1 cells to homeostatic- and antigen-induced signals and thus contribute to the functional heterogeneity among these innate-like B cells. PMID:27667991

  13. Rearrangement and expression of endogenous immunoglobulin genes occur in many murine B cells expressing transgenic membrane IgM.

    PubMed

    Stall, A M; Kroese, F G; Gadus, F T; Sieckmann, D G; Herzenberg, L A; Herzenberg, L A

    1988-05-01

    Transgenic mice carrying immunoglobulin genes coding for mu heavy chain and kappa light chain have been used to study the mechanisms involved in allelic and isotypic exclusion. We report here that individual cells from transgenic mice carrying a functionally rearranged mu heavy chain gene (capable of generating both membrane and secreted forms of IgM) can rearrange an endogenous mu heavy chain gene and simultaneously produce both transgenic and endogenous IgM. These "double-producing" cells express both endogenous and transgenic IgM in the cytoplasm (detected by immunohistology) and on the cell surface (detected by multiparameter fluorescence-activated cell sorter analysis). In addition, they secrete mixed IgM molecules containing both transgenic and endogenous mu heavy chains (detected in serum by radioimmune assay). The transgenic mice studied also have relatively large numbers of cells that produce endogenous immunoglobulin in the absence of detectable transgenic immunoglobulin ("endogenous-only cells"). The mechanisms that generate double-producing cells and endogenous-only cells appear to be under genetic control because the frequencies of these B-cell populations are characteristic for a given transgenic line. Thus, our findings indicate that more is involved in triggering allelic exclusion than the simple presence or absence of membrane mu heavy chains (as has been previously postulated).

  14. Elucidation of the potential disease-promoting influence of IgM apoptotic cell-reactive antibodies in lupus.

    PubMed

    Malik, M; Arora, P; Sachdeva, R; Sharma, L; Ramachandran, V G; Pal, R

    2016-06-01

    The undigested remnants of apoptosis are believed to stimulate the generation of autoantibodies in lupus. The biological properties of initiator, disease-specific IgM antibodies that specifically recognize apoptotic cells, readily detected in the sera of lupus patients, remain unclear. Apoptotic cell-reactive IgM monoclonal antibodies (generated from lupus-prone mice), as opposed to control IgM, preferentially stimulated maturation of bone marrow-derived dendritic cells (BMDCs) derived from such mice, relative to BMDCs derived from healthy mice. An influence of both antibody specificity and cell genotype was also apparent in the secretion of signature inflammatory cytokines. Immunization of such antibodies in lupus-prone animals induced increases in total serum IgG levels, with the elicited antibodies also preferentially recognizing moieties on dying cells. An expanded specificity was apparent both upon Western blot on cellular lysate and from the enhanced recognition of dsDNA, Ro60, RNP68 and Sm; the antibody most efficient in mediating autoreactive diversity, while being germline encoded, also induced the highest degree of phenotypic changes on BMDCs. Apoptotic cell-reactive IgM antibodies may therefore be potentially capable of influencing the course of systemic autoimmune disease by affecting both innate and adaptive immunity.

  15. Serologic cross-reactivity of human IgM and IgG antibodies to five species of Ebola virus.

    PubMed

    Macneil, Adam; Reed, Zachary; Rollin, Pierre E

    2011-06-01

    Five species of Ebola virus (EBOV) have been identified, with nucleotide differences of 30-45% between species. Four of these species have been shown to cause Ebola hemorrhagic fever (EHF) in humans and a fifth species (Reston ebolavirus) is capable of causing a similar disease in non-human primates. While examining potential serologic cross-reactivity between EBOV species is important for diagnostic assays as well as putative vaccines, the nature of cross-reactive antibodies following EBOV infection has not been thoroughly characterized. In order to examine cross-reactivity of human serologic responses to EBOV, we developed antigen preparations for all five EBOV species, and compared serologic responses by IgM capture and IgG enzyme-linked immunosorbent assay (ELISA) in groups of convalescent diagnostic sera from outbreaks in Kikwit, Democratic Republic of Congo (n=24), Gulu, Uganda (n=20), Bundibugyo, Uganda (n=33), and the Philippines (n=18), which represent outbreaks due to four different EBOV species. For groups of samples from Kikwit, Gulu, and Bundibugyo, some limited IgM cross-reactivity was noted between heterologous sera-antigen pairs, however, IgM responses were largely stronger against autologous antigen. In some instances IgG responses were higher to autologous antigen than heterologous antigen, however, in contrast to IgM responses, we observed strong cross-reactive IgG antibody responses to heterologous antigens among all sets of samples. Finally, we examined autologous IgM and IgG antibody levels, relative to time following EHF onset, and observed early peaking and declining IgM antibody levels (by 80 days) and early development and persistence of IgG antibodies among all samples, implying a consistent pattern of antibody kinetics, regardless of EBOV species. Our findings demonstrate limited cross-reactivity of IgM antibodies to EBOV, however, the stronger tendency for cross-reactive IgG antibody responses can largely circumvent limitations in the

  16. Heat inactivation can differentiate between IgG and IgM antibodies in the pretransplant cross match.

    PubMed

    Al-Muzairai, I A; Mansour, M; Almajed, L; Alkanderi, N; Alshatti, N; Samhan, M

    2008-09-01

    The presence of IgG antibodies in the pretransplant cross-match (XM) test results in hyperacute rejection, but IgM antibodies are inconsequential. The XM should be able to differentiate between IgG and IgM antibodies. This study evaluated 3 methods. This study was based on 500 patients for whom XM were performed between 2004 and 2006 with all 3 techniques. Two patient sera were used: normal serum and heat inactivated serum, which was prepared by incubating patient serum at 63 degrees C for 10 minutes to destroy IgM antibodies. The efficiencies of flow cytometry XM (FC-XM), dithiothreitol complement-dependent microlymphocytotoxicity (DTT/CDC-XM), and heat inactivation (HI-CDC-XM) to differentiate between IgG and IgM were evaluated by using both sera. Patients with positive XM, and negative HI-CDC-XM were reported as negative XM. During the study period, there were 70 patients with positive B-cell XM. Forty-nine became negative after HI-XM, and 21 remained positive. Only 34 cases became negative after DTT-CDC-XM and 36 remained positive. HI-CDC-XM was comparable to FC-XM; all patients testing negative with this technique experienced successful renal transplantations without hyperacute, accelerated, or acute rejection episodes. Our study showed that HI-CDC-XM was effective at exclude donor-specific IgM antibodies, a result which was comparable to FCXM to detect only IgG antibodies. HI is simple and rapid and does not involve any extra equipment or cost.

  17. Clinical evaluation of dengue RNA, NS1, and IgM for diagnosis of dengue in Southern China.

    PubMed

    Chen, Xinliang; Chen, Rui; Gu, Wenshen; He, Jian; Cai, Weipeng; Li, Jiajia; Duan, Chaohui; Yan, Haiyan

    2016-01-01

    In 2014, a large outbreak of dengue occurred in Guangzhou, China. This outbreak prompted us to evaluate NS1 and RNA for the early diagnosis of acute dengue infection, in addition to the combination with IgM antibody. We aimed to find the differences of three assays about dengue diagnosis. This study was an evaluation of diagnosis test. Based on WHO criteria 2009, dengue RNA, NS1, and IgM/IgG were detected from 294 patients (180 dengue patients, 114 non-dengue patients) by three diagnostic kits made in China. The χ(2) test, sensitivity, and specificity were used in statistical analysis. The ratios of dengue patients with low platelet counts (<100 × 10(9) /L 32.2%) or white blood cell counts (<4.0 × 10(9) /L 58.9%) were significantly higher compared to non-dengue patients (P < 0.05). Dengue NS1 was shown sensitive (93.9%) for diagnostic use. RNA had a better performance with 98.1% of sensitivity from day 1 to day 4 after illness onset. IgM performed better at day 5 or more with 74.0% of sensitivity. The diagnostic rate using a combination of RNA and IgM was 97.8% and 96.7% using NS1 and IgM. A patient with low platelet and white blood cell counts needs additional tests for dengue during an epidemic. RNA and NS1 were most valuable for early diagnosis of dengue, whereas IgM was best suited as a supplementary method for patients at day 5 or more after illness onset.

  18. Cross-reactions in IgM ELISA tests to Legionella pneumophila sg1 and Bordetella pertussis among children suspected of legionellosis; potential impact of vaccination against pertussis?

    PubMed

    Pancer, Katarzyna Wanda

    2015-01-01

    The objective of this study was preliminary evaluation of IgM cross-reaction in sera collected from children hospitalized because of suspected legionellosis. Sera with positive IgM results to L. pneumophila sgs1-7, B. pertussis or with simultaneous detection of IgM antibodies to L. pneumophila sgs1-7 and B. pertussis, or IgM to L. pneumophila sgs1-7 and M. pneumoniae in routine tests, were selected. In total, an adapted pre-absorption test was used for the serological confirmation of legionellosis in the sera of 19 children suspected of legionellosis, and also in 3 adult persons with confirmed Legionnaires' disease. Sera were pre-absorbed with antigens of L. pneumophila sg1, B. pertussis or both, and tested by ELISA tests. The reduction of IgM antibody level by pre-absorption with antigen/antigens was determined. Reduction of anti-Lpsgs1-7 IgM by pre-absorption with L.pneumophila sg1 antigen ranged from 1.5 to 80, and reduction of anti-Bp IgM by pre-absorption with B. pertussis ranged from 2.0 to 23.8. Reduction by both antigens varied depending on the age of the patients: among children <4 yrs.old, the reduction of anti-B. pertussis IgM by both antigens was higher than for B. pertussis antigen alone. Based on the high difference (≥ 2 times) between reduction by L.pneumophila sg1 and by B. pertussis antigen, legionellosis was confirmed in 8/19 children. The majority of them also indicated IgM positive/borderline results for B. pertussis or M.pneumoniae in routine ELISA tests. As a preliminary, we posed a hypothesis of a potential impact of an anti-pertussis vaccination on the results obtained in anti-L. pneumophila ELISA IgM tests among young children.

  19. Studies on the structural and biological functions of the Cμ3 and Cμ4 domains of IgM

    PubMed Central

    Bubb, M. O.; Conradie, J. D.

    1978-01-01

    The development of methods for the production of intact Cμ3 and Cμ4 domains of IgM have made possible the assessment of some of their structural and biological functions. Antiserum against Fcμ fragment detected both domains and illustrated their complete antigenic non-identity. Circular dichroism spectroscopy and the retention of antigenicity indicated that both domains had retained most of their native structure. No interaction of the type Cμ3—Cμ3, Cμ4—Cμ4 or Cμ3—Cμ4 could be detected under non-dissociating conditions by analytical ultracentrifugation or molecular exclusion chromatography experiments. These results lead us to believe that the transmission of effector messages between the Fab and Fc parts of IgM takes place through structural changes at the quaternary level. C[unk]1-fixation experiments with IgM and several of its fragments and domains show that (a) the Cμ4 domain contains the C[unk]1-fixing site; (b) the high C[unk]1-fixing capacity of IgM or Fc5μ cannot be explained on the basis of a simple accumulative model of complement fixing domains; (c) the C[unk]1-fixing site is independent of the native structure of the Cμ4 domain; (d) the C[unk]1-fixing site does not contain carbohydrate. Examination of the IgM receptor on the surface of human T lymphocytes show that (a) Cμ4 domain is primarily responsible for the reaction and Cμ3 domain has very little affinity; (b) native structure is essential for the reaction because reduction and alkylation of the Cμ4 domain destroyed both its original conformation and affinity for this receptor; (c) IgM and Fc5μ had a much greater affinity for the receptor than monomeric subunits: (d) carbohydrate on Cμ4 domain is not involved in the affinity reaction. ImagesFigure 1Figure 2 PMID:417020

  20. GD3/proteosome vaccines induce consistent IgM antibodies against the ganglioside GD3.

    PubMed

    Livingston, P O; Calves, M J; Helling, F; Zollinger, W D; Blake, M S; Lowell, G H

    1993-09-01

    The gangliosides of melanoma and other tumours of neuroectodermal origin are suitable targets for immune intervention with tumour vaccines. The optimal vaccines in current use contain ganglioside plus bacillus Calmette-Guérin and induce considerable morbidity. We have screened a variety of new adjuvants in the mouse, and describe one antigen-delivery system, proteosomes, which is especially effective. Highly hydrophobic Neisserial outer membrane proteins (OMP) form multimolecular liposome-like vesicular structures termed proteosomes which can readily incorporate amphiphilic molecules such as GD3 ganglioside. The optimal GD3/proteosome vaccine formulation for induction of GD3 antibodies in the mouse is determined. Interestingly, the use of potent immunological adjuvants in addition to proteosomes augments the IgM and IgG antibody titres against OMP in these vaccines but GD3 antibody titres are unaffected. The application of proteosomes to enhance the immune response to GD3 extends the concept of the proteosome immunopotentiating system from lipopeptides to amphipathic carbohydrate epitopes such as cell-surface gangliosides. The demonstrated safety of meningococcal OMP in humans and the data in mice presented here suggest that proteosome vaccines have potential for augmenting the immunogenicity of amphipathic tumour antigens in humans.

  1. Radio resource management for CDMA systems with interference guard margin (IGM)

    NASA Astrophysics Data System (ADS)

    Chen, Huan; Kumar, Sunil; Kuo, C.-C. Jay

    2001-11-01

    A call admission control (CAC) scheme and a resource reservation estimation (RRE) method suitable for the wide-band code division multiple access (W-CDMA) systems are proposed in this work. The proposed CAC scheme gives preferential treatment to high priority calls, such as handoff calls, by pre-reserving a certain amount of channel margin against the interference effect. It is called the interference guard margin (IGM) scheme. The amount of guard margin is determined by the measurement performed by the RRE module in base stations. Each RRE module dynamically adjusts the level of guard margin by referencing traffic conditions in neighboring cells based upon users' requests. A comprehensive service model is adopted to accommodate the scenario of multiple services supported in the W-CDMA system. The service model of consideration includes not only mobile terminal's service rate (source rate) but also different levels of priorities, mobility and rate adaptivity characteristics. Simulations are conducted with OPNET to study the performance of the proposed scheme in term of the objective function under different traffic conditions.

  2. IgM natural autoantibodies against bromelain-treated mouse red blood cells recognise carbonic anhydrase.

    PubMed

    Jonusys, A M; Cox, K O; Steele, E J

    1991-01-01

    Carbonic anhydrase (CA) from mouse erythrocyte membranes is recognised as an autoantigen in Western blotting experiments with FUB 1, a murine IgM monoclonal antibody that binds both phosphatidylcholine and bromelain-treated mouse red blood cells (BrMRBC). Serum from mice stimulated with lipopolysaccharide (LPS-serum) also recognises CA. From SDS-PAGE, and blotting experiments with whole mouse erythrocytes, we found two closely spaced glycoprotein bands in the 30 kD region that reacted with both FUB 1 and LPS-serum. One of the molecular weight markers, bovine carbonic anhydrase which is of a molecular weight of about 30 kD, electrophoresed in the same 30 kD region also reacted with these antibodies. Carbonic anhydrases from a range of mammalian species were found to be crossreactive with FUB 1 and LPS-serum by Western blotting, whereas human glycophorin A and human asialoglycophorin were not recognised by the antibodies. FUB 1 specifically recognises both native and denatured bovine carbonic anhydrase in ELISA assays. The serological identity of the determinants of CA and BrMRBC was confirmed by specific absorption of both FUB 1 and LPS-serum with BrMRBC and normal mouse erythrocytes. We propose that a native autoantigenic epitope on erythrocytes may be revealed by the proteolytic action of bromelain and that this determinant is associated, at least in part, with carbonic anhydrase.

  3. Physical biochemical properties of IgM from a teleost fish.

    PubMed Central

    Mochida, K; Lou, Y H; Hara, A; Yamauchi, K

    1994-01-01

    Teleost fish are able to produce IgM class antibody, as are other vertebrates. When the teleost fish Oreochromis niloticus was immunized with bovine serum albumin (BSA), it produced antibody to BSA with an average avidity of 7.4 x 10(8)/M. Thus, dissociation of antigen-antibody complexes only occurred at conditions of < pH 2.5, > pH 11, > 4M NaI or > 4M urea, demonstrating high stability of the complex. Western blot analyses further showed the high specificity of the antibody to BSA. In contrast to mammals, when the fish was challenged with multiple protein antigens, it produced antibody only to the major component but not to others. The antibody generated to a specific antigen accounted for up to 1.1% of whole serum protein or 7.0% of whole immunoglobulin. We conclude (1) the systemic antibody response in teleost fish may be an 'all or nothing' response, which is different from that in mammals; (2) the quality (specificity and affinity) of the antibody produced is similar to that of mammals. The findings not only reveal a quite different strategy of immune response in fish, but also raise the possibility of technical application. Images Figure 2 Figure 3 PMID:7875748

  4. Deterioration of lung function is associated with presence of IgM rheumatoid factor and smoking in patients with systemic sclerosis.

    PubMed

    Broholm, Berit; Ullman, Susanne; Halberg, Poul; Jacobsen, Søren

    2008-08-01

    Smoking is a known risk factor for the development of several lung diseases, autoimmune diseases, and IgM rheumatoid factor (RF) in nonrheumatic persons. In patients with rheumatoid arthritis and IgM RF the diffusion capacity is decreased in smokers but not in nonsmokers. In the present study of patients with systemic sclerosis (SSc) the influence of smoking and IgM RF on the lung function was calculated. One hundred fifty-five persons with SSc had vital capacity (VC) and diffusing capacity (DLco) measured at least twice with at least 1-year interval as percents of predicted values according to gender, age, height, and weight. The yearly changes in VC and DLco were calculated, Delta VC and Delta DLco, respectively. IgM RF was measured at the beginning of the study. Smoking was defined as having ever smoked. Statistically significant deterioration of VC and DLco in patients with circulating IgM RF was found only in smokers or previous smokers, P = 0.007 and P = 0.01, respectively. These findings were confirmed by means of multiple regression analyses. The presence of IgM RF in smoking SSc patients is associated with deteriorating lung function. Whether this is a causal association and whether the presence of IgM RF in smoking patients with SSc actually confers an increased risk of pulmonary damage remains to be determined.

  5. [Detection of rubella specific IgM on gel filtration through Sephadex G 200: use of dithiothreitol and limits with MnCl2-heparine pretreatment (author's transl)].

    PubMed

    Freymuth, F; Daon, F; Vergnaud, M; Valdazo, A

    1979-01-01

    The detection of rubella haemagglutination inhibiting antibody, in the IgM fraction of the serum, on gel filtration through Sephadex G 200, needs precautions to exclude false results. Treatment with dithiothreitol is a satisfactory method for confirming the content of rubella IgM antibody. The failure of MnCl2-heparin pretreatment to remove non specific inhibitors of rubella hemagglutinin is unfrequent (7/108) and so do be repeated. Rarely (1/108) aggregated IgG fractionates with IgM and yield false positive results. PMID:475077

  6. Toxocara canis glycans influence antigen recognition by mouse IgG1 and IgM antibodies.

    PubMed

    Długosz, Ewa; Wiśniewski, Marcin

    2016-01-01

    The impact of sugar moieties of Toxocara canis glycoprotein antigens on their recognition by infected mouse antibodies was investigated in this study. Native TES and recombinant Toxocara mucins generated in Pichia pastoris yeast as well as their deglycosylated forms were used in ELISA. TES and recombinant mucins were equally recognized by T. canis infected mouse IgG1 antibodies. IgM immunoglobulins predominantly recognized TES antigens. Among mucins recognition of Tc-MUC-4 was the most significant. Deglycosylation of antigens resulted in significant loss of IgM and IgG1 reactivity to TES, mucins, Tc-MUC-3 and Tc-MUC-4. The presence of sugar moieties had no influence on IgE binding to native or recombinant T. canis antigens. Our results suggest that glycans are involved in epitope formation what should be taken into consideration in production of recombinant helminth antigens for diagnostic purposes. PMID:26751891

  7. Improved surveillance of Japanese encephalitis by detection of virus-specific IgM in desiccated blood specimens*

    PubMed Central

    Burke, D. S.; Chatiyanonda, K.; Anandrik, S.; Nakornsri, S.; Nisalak, A.; Hoke, C. H.

    1985-01-01

    An IgM antibody-capture type enzyme-linked immunoassay (MAC ELISA) was compared with the haemagglutination inhibition method (HI) for establishing a laboratory diagnosis of acute Japanese encephalitis (JE) virus infection using specimens of dried blood eluted from filter paper strips. Paired samples from 243 encephalitis patients, which had been obtained by mail through a national surveillance programme in Thailand, were tested. During the peak of the 1983 encephalitis epidemic, 72% of cases were diagnosed as Japanese encephalitis by MAC ELISA, compared with only 38% by HI. During non-epidemic periods, the proportions diagnosed as Japanese encephalitis by MAC ELISA or HI were 26% and 33%, respectively. Detection of IgM anti-JE activity by the antibody-capture immunoassay is superior to the HI method for establishing a diagnosis of acute Japanese encephalitis using dried blood specimens. PMID:3011302

  8. Increase in Anti-Gal IgM Level is Associated With Early Graft Failure in Intraportal Porcine Islet Xenotransplantation

    PubMed Central

    Lee, Haneulnari; Park, Eun Mi; Kim, Jong Min; Shin, Jun-Seop; Kim, Jung-Sik; Park, Chung-Gyu; Kim, Sang Joon

    2015-01-01

    Background Anti-Gal is a major antibody induced in non-human primates (NHPs) after xenotransplantation. To understand the mechanism of graft rejection, we investigated the association between anti-Gal responses and graft failure in NHP recipients of porcine islet transplantation (PITx). Methods Intraportal PITx was performed in 35 diabetic NHPs, and graft function was monitored. Early graft failure (EGF) was defined as loss of graft function within a month after PITx. Seven, 19, nine NHPs received immunosuppression (IS) without CD40 pathway blockade (Group I), with anti-CD154 (Group II), and with anti-CD40 (Group III), respectively. The anti-Gal levels on day 0 and day 7 of PITx were measured by ELISA. Results The frequency of EGF was significantly lower in Group II (26.3%) than in Group I (100%, P=0.0012) and Group III (77.8%, P=0.0166). While levels of anti-Gal IgG in Group I and anti-Gal IgM in Group III increased on day 7 compared with day 0 (P=0.0156 and 0.0273), there was no increase in either on day 7 in Group II. The ratio of anti-Gal IgM or IgG level on day 7 to that on day 0 (Ratio7/0) was significantly higher in recipients with EGF than without EGF (P=0.0009 and 0.0027). ROC curve analysis of anti-Gal IgM Ratio7/0 revealed an area under the curve of 0.789 (P=0.0003). Conclusions IS with anti-CD154 suppressed anti-Gal responses and prevented EGF in PITx. Anti-Gal IgM Ratio7/0, being associated with EGF, is a predictive marker for EGF. PMID:26354349

  9. DC-SIGN–expressing macrophages trigger activation of mannosylated IgM B-cell receptor in follicular lymphoma

    PubMed Central

    Amin, Rada; Mourcin, Frédéric; Uhel, Fabrice; Pangault, Céline; Ruminy, Philippe; Dupré, Loic; Guirriec, Marion; Marchand, Tony; Fest, Thierry; Lamy, Thierry

    2015-01-01

    Follicular lymphoma (FL) results from the accumulation of malignant germinal center (GC) B cells leading to the development of an indolent and largely incurable disease. FL cells remain highly dependent on B-cell receptor (BCR) signaling and on a specific cell microenvironment, including T cells, macrophages, and stromal cells. Importantly, FL BCR is characterized by a selective pressure to retain surface immunoglobulin M (IgM) BCR despite an active class-switch recombination process, and by the introduction, in BCR variable regions, of N-glycosylation acceptor sites harboring unusual high-mannose oligosaccharides. However, the relevance of these 2 FL BCR features for lymphomagenesis remains unclear. In this study, we demonstrated that IgM+ FL B cells activated a stronger BCR signaling network than IgG+ FL B cells and normal GC B cells. BCR expression level and phosphatase activity could both contribute to such heterogeneity. Moreover, we underlined that a subset of IgM+ FL samples, displaying highly mannosylated BCR, efficiently bound dendritic cell–specific intercellular adhesion molecule-3–grabbing nonintegrin (DC-SIGN), which could in turn trigger delayed but long-lasting BCR aggregation and activation. Interestingly, DC-SIGN was found within the FL cell niche in situ. Finally, M2 macrophages induced a DC-SIGN–dependent adhesion of highly mannosylated IgM+ FL B cells and triggered BCR-associated kinase activation. Interestingly, pharmacologic BCR inhibitors abolished such crosstalk between macrophages and FL B cells. Altogether, our data support an important role for DC-SIGN–expressing infiltrating cells in the biology of FL and suggest that they could represent interesting therapeutic targets. PMID:26272216

  10. Natural and adaptive IgM antibodies in the recognition of tumor-associated antigens of breast cancer (Review)

    PubMed Central

    DÍAZ-ZARAGOZA, MARIANA; HERNÁNDEZ-ÁVILA, RICARDO; VIEDMA-RODRÍGUEZ, RUBÍ; ARENAS-ARANDA, DIEGO; OSTOA-SALOMA, PEDRO

    2015-01-01

    For early detection of cancer, education and screening are important, but the most critical factor is the development of early diagnostic tools. Methods that recognize the warning signs of cancer and take prompt action lead to an early diagnosis; simple tests can identify individuals in a healthy population who have the disease but have not developed symptoms. Early detection of cancer is significant and is one of the most promising approaches by which to reduce the growing cancer burden and guide curative treatment. The early diagnosis of patients with breast cancer is challenging, since it is the most common cancer in women worldwide. Despite the advent of mammography in screening for breast cancer, low-resource, low-cost alternative tools must be implemented to complement mammography findings. IgM is part of the first line of defense of an organism and is responsible for recognizing and eliminating infectious particles and removing transformed cells. Most studies on breast cancer have focused on the development of IgG-like molecules as biomarkers or as a treatment for the advanced stages of cancer, but autoantibodies (IgM) and tumor-associated antigens (proteins or carbohydrates with aberrant structures) have not been examined as early diagnostic tools for breast cancer. The present review summarizes the function of natural and adaptive IgM in eliminating cancer cells in the early stages of pathology and their value as early diagnostic tools. IgM, as a component of the immune system, is being used to identify tumor-associated antigens and tumor-associated carbohydrate antigens. PMID:26133558

  11. Recombinant carp IL-4/13B stimulates in vitro proliferation of carp IgM(+) B cells.

    PubMed

    Yamaguchi, Takuya; Miyata, Shunsuke; Katakura, Fumihiko; Nagasawa, Takahiro; Shibasaki, Yasuhiro; Yabu, Takeshi; Fischer, Uwe; Nakayasu, Chihaya; Nakanishi, Teruyuki; Moritomo, Tadaaki

    2016-02-01

    Teleost IL-4/13B is a cytokine related to mammalian IL-4 and IL-13, of which hitherto the function had not been studied at the protein level. We identified an IL-4/13B gene in common carp (Cyprinus carpio) and expressed the recombinant protein (rcIL-4/13B). RcIL-4/13B was shown to stimulate proliferation of IgM(+) B cells, because after four days of stimulation the IgM(+) fraction of carp kidney and spleen leukocytes had formed many cell colonies, whereas such colonies were not found in the absence of rcIL-4/13B stimulation. After nine days of incubation with rcIL-4/13B these cells had proliferated to more than 3-to-7-fold higher numbers when compared to untreated cells. The proliferating cells contained a majority of IgM(+) cells but also other cells, as indicated by FACS and RT-PCR analyses. The important conclusion is that in fish not only IL-4/13A has B cell stimulating properties, as a previous publication has shown, but also IL-4/13B. PMID:26766176

  12. Rapid diagnosis of echovirus type 33 meningitis by specific IgM detection using an enzyme linked immunosorbent assay (ELISA).

    PubMed

    Chomel, J J; Thouvenot, D; Fayol, V; Aymard, M

    1985-01-01

    During an outbreak of meningitis in France (in the Lyon area), from June to October 1982, serum and stool samples were collected from 227 patients. An enzyme-linked immunosorbent assay (ELISA) for titrating IgG and IgM antibodies anti-echovirus type 33 was developed and compared with the virus isolation technique, and with the titration of neutralizing antibodies. In 39 patients excreting echovirus 33 in faeces, the ELISA test allowed a positive serodiagnosis in 85% of the cases by detection of specific IgM (64% of the cases) and by seroconversion (21%). Compared with the neutralization (Nt) test, ELISA was found to be more sensitive. The antibody titres in ELISA were over 50 times higher and detected earlier than the neutralizing antibodies. This early immune response allowed a rapid diagnosis by specific IgM detection in the acute sera collected within 8 days after the appearance of the clinical symptoms in more than 50% of the 97 patients examined, whereas the Nt test allowed a positive serodiagnosis in only 32% of the patients. The use of a caesium chloride purified antigen insured the specificity of the reactions.

  13. Use of monoclonal antibodies in an ELISA to detect IgM class antibodies specific for Toxoplasma gondii.

    PubMed Central

    Balfour, A H; Harford, J P; Goodall, M

    1987-01-01

    Two monoclonal antibodies CH6 and C1E3 were used in an antibody class capture assay for the detection of IgM antibodies specific for Toxoplasma gondii. CH6 was used on the solid phase to capture human IgM. After a Toxoplasma gondii antigen had been added, specifically bound material was detected using C1E3 coupled to horseradish peroxidase. The assay was compared with an established system using polyclonal antisera at both the capture and antigen detection stages. A good correlation was found, with 97.3% (125 of 128) of sera giving the same classification in both assays. Three sera were positive only in the polyclonal system. No false positive results were found when 118 negative sera were examined. The two monoclonal antibodies provide a viable alternative to the use of polyclonal sera at the capture and antigen detection stages in the antibody class capture assay for the measurement of specific IgM against T gondii. PMID:3654986

  14. Amyloid-like IgM deposition neuropathy: a distinct clinico-pathologic and proteomic profiled disorder

    PubMed Central

    Figueroa, Juan J.; Bosch, E. Peter; Dyck, P. James B.; Singer, Wolfgang; Vrana, Julie A.; Theis, Jason D.; Dogan, Ahmet; Klein, Christopher J.

    2014-01-01

    Some patients with immunoglobulin paraproteinemic neuropathy have intra-nerve deposits that morphologically mimick amyloid, but do no stain with Congo red. Patients with amyloid-like deposits were identified. The nerve amyloid-like aggregates were studied by laser microdissection and dual mass spectrometry. Three male patients, all with IgM gammopathy, and neuropathy were identified. Follow-up, disease duration was 5, 19, and 7 years, respectively. All had progressive asymmetric sensory-onset distal axonal polyneuropathy with late motor involvement. Autonomic symptoms occurred in only one after 13 years of symptoms. None had clinical cardio-renal involvement. One had skin papules with dermal amyloid-like deposits. Endoneurial amyloid-like deposits had granulo-fibrillar ultrastructure. Mass spectrometry of laser-dissected deposits identified IgM pentameric macroglobulin (heavy, light, and joining chains) without amyloid-associated proteins including absent apolipoprotein E and serum amyloid P-component. Amyloid-like neuropathy has distinct clinical, pathologic, and proteomic features which expand the spectrum of IgM neuropathies. Patients have favorable survival, relative absence of autonomic features, and distinct proteomic profiles of the infiltrative protein in nerve. PMID:22734903

  15. ALS Association

    MedlinePlus

    ... toward a world without ALS! Walk to Defeat ALS® Walk to Defeat ALS® draws people of all ... We need your help. I Will Advocate National ALS Registry The National ALS Registry is a congressionally ...

  16. ABO (histo) blood group phenotype development and human reproduction as they relate to ancestral IgM formation: A hypothesis.

    PubMed

    Arend, Peter

    2016-01-01

    The formation of a histo (blood) group) ABO phenotype and the exclusion of an autoreactive IgM or isoagglutinin activity arise apparently in identical glycosylation of complementary domains on cell surfaces and plasma proteins. The fundamental O-glycan emptiness of the circulating IgM, which during the neonatal amino acid sequencing of the variable regions is exerting germline-specific O-GalNAc glycan-reactive serine/threonine residues that in the plasma of the adult human blood group O individuals apparently remain associated with the open glycosidic sites on the ABOH convertible red cell surface, must raise suggestions on a transient expression of developmental glycans, which have been "lost" over the course of maturation. In fact, while the mammalian non-somatic, embryogenic stem cell (ESC)- germ cell (GC) transformation is characterized by a transient and genetically as-yet-undefined trans-species-functional O-GalNAc glycan expression, in the C57BL/10 mouse such expression was potentially identified in growth-dependent, blood group A-like GalNAc glycan-bearing, ovarian glycolipids complementary with the syngeneic anti-A reactive IgM, which does not appear in early ovariectomized animals. This non-somatically encoded, polyreactive, ancestral IgM molecule has not undergone clonal selection and does primarily not differentiate between self and non-self and might, due to amino acid hydroxyl groups, highly suggest substrate competition with subsequent O-glycosylations in ongoing ESC-GC transformations and affecting GC maturation. However, the membrane-bound somatic N/O-glycotransferases, which initiate, after formation of the zygote, the complex construction of the human ABO phenotypes in the trans cisternae of the Golgi apparatus, are associated and/or completed with soluble enzyme versions exerting identical specificities in plasma and likely competing vice versa by glycosylation of neonatal IgM amino acids, where they suggest to accomplish the clearance of anti

  17. Human monoclonal IgM with autoantibody activity against two gangliosides (GM1 and GD1b) in a patient with motor neuron syndrome.

    PubMed Central

    Jauberteau, M O; Gualde, N; Preud'Homme, J L; Rigaud, M; Gil, R; Vallat, J M; Baumann, N

    1990-01-01

    Small amounts of oligoclonal immunoglobulins were detected by Western blotting in the serum from a patient with motor neuron syndrome. The prominent one, a monoclonal IgM lambda, reacted strongly with the gangliosides GM1 and GD1b and more weakly with asialo GM1, as shown by immunoenzymatic staining of thin-layer chromatograms of gangliosides, ELISA on purified glycolipid coats and immunoadsorption with purified GM1. Affinity-chromatography with purified GM1 resulted in the purification of monoclonal IgM lambda. This purified IgM and its Fab fragments showed the same pattern of reactivity with gangliosides as that observed with whole serum. Such monoclonal IgM could be responsible for motor neuron diseases in some patients with overt or barely detectable monoclonal gammopathies. Images Fig. 2 Fig. 3 PMID:2357844

  18. Fetal lambs are depleted of IgM+ cells following a single injection of an anti-IgM antibody early in gestation.

    PubMed Central

    Press, C M; Reynolds, J D; McClure, S J; Simpson-Morgan, M W; Landsverk, T

    1996-01-01

    B-cell depleted fetal sheep were created following a single injection of an anti-IgM monoclonal antibody early in gestation. Six sheep fetuses were given a single intraperitoneal injection of a monoclonal antibody directed against IgM at 63 days of gestation (gestation in sheep = 150 days). The fetuses were killed at 138-142 days of gestation and lymphoid tissues were collected for subsequent light microscopy and immunohistochemical examination. The ileal and jejunal Peyer's patch (PP) follicles in four of the six injected fetuses were markedly reduced in size. Cells in the rudimentary follicles of the ileal PP of these animals showed no reactivity for IgM and most were negative for CD45. The dome regions contained many T cells, which were predominantly CD8+ cells and included gamma delta T cells. The interfollicular areas of the PP of the markedly affected fetuses contained large populations of T cells. The spleen and lymph nodes were also markedly depleted of IgM+ cells and these tissues contained only a small, scattered population of weakly IgM+ cells. Follicular accumulations of IgM+ cells were absent. Large populations of T cells were present in the white pulp of the spleen and cortex of the lymph nodes. The liver did not contain IgM+ cells and the medulla of the thymus was depleted of IgM+ cells. The results of this study suggest that a surface IgM+ B-cell population is present in the sheep fetus at 63 days of gestation, which is essential for the colonization of the ileal PP and subsequent B-cell development. Images Figure 1 Figure 2 Figure 3 Figure 4 PMID:8707346

  19. The use of IgM anti-D coated cells in the deliberate immunisation of Rh-negative male volunteers.

    PubMed

    Lee, D; Flowerday, M H; Tomlinson, J

    1977-01-01

    Attempts to produce anti-D in rhesus-negative men using reconstituted frozen rhesus-positive cells were unsuccessful. The use of cells coated with IgM anti-D for immunisation stimulates anti-D production, and a comparison between two groups, one having cells coated with IgM anti-D, and the other having identical cells without coating suggests that there is an enhancement of the immune response when IgM-coated cells are used.

  20. Pregnancy Outcomes of Mothers with Detectable CMV-Specific IgM Antibodies: A Three-Year Review in a Large Irish Tertiary Referral Maternity Hospital

    PubMed Central

    Drew, Richard J.; Stapleton, Patrick; Abu, Hala; Healy, Eibhlín; Ferguson, Wendy; De Gascun, Cillian; O'Gorman, Joanne; Eogan, Maeve

    2015-01-01

    A retrospective audit was performed for all obstetric patients who had positive CMV IgM results between January 2012 and December 2014 in the Rotunda Hospital, Ireland. In total, 622 CMV IgM positive tests were performed on samples from 572 patients. Thirty-seven patients had a positive CMV IgM result (5.9%) on the Architect system as part of the initial screening. Three patients were excluded as they were not obstetric patients. Of the 34 pregnant women with CMV IgM positive results on initial screening, 16 (47%) had CMV IgM positivity confirmed on the second platform (VIDAS) and 18 (53%) did not. In the 16 patients with confirmed positive CMV IgM results, four (25%) had acute infection, two (12.5%) had infection of uncertain timing, and ten (62.5%) had infection more than three months prior to sampling as determined by the CMV IgG avidity index. Two of the four neonates of women with low avidity IgG had CMV DNA detected in urine. Both these cases had severe neurological damage and the indication for testing their mothers was because the biparietal diameter (BPD) was less than the 5th centile at the routine 20-week gestation anomaly scan. PMID:26696757

  1. Biodistribution of 111In-labelled IgG and IgM in experimental infection.

    PubMed

    Oyen, W J; Boerman, O C; Subramanian, R; Koenders, E B; Claessens, R A; van der Meer, J W; Corstens, F H

    1996-07-01

    Both the protein used and the conjugation method are factors which may be relevant for targeting infection with 111In-labelled proteins. In this study, human immunoglobulin G (IgG), conjugated to either DTPA or LiLo, and LiLo conjugated human immunoglobulin M (IgM) were evaluated. In rats with Staphylococcus aureus calf muscle infection, biodistribution was determined 6, 24 and 48 h after the injection of 111In-DTPA-IgG, 111InLiLo-IgG or 111In-LiLo-IgM. Absolute abscess uptake of 111In-LiLo-IgG was significantly higher than that of 111In-DTPA-IgG (P < 0.05). Since blood clearance of 111In-LiLo-IgG was initially significantly slower (P < 0.01), the higher abscess uptake did not result in higher abscess-to-background ratios. 111In-LiLo-IgG accumulated to a greater extent in the liver (P < 0.001). 111In-DTPA-IgG showed higher uptake in the kidneys and bone marrow (P < 0.001 and P < 0.01, respectively). Although decreasing over time, 111In-LiLo-IgM showed reasonable abscess uptake and rapid blood clearance, resulting in higher abscess-to-background ratios compared with 111In-LiLo-IgG (P < 0.01). However, liver and spleen uptake were three- to four-fold higher than that of 111In-LiLo-IgG (P < 0.001). Compared with DTPA-conjugation, chelation with LiLo has a minor influence on abscess targeting of 111In-labelled IgG. However, differences in blood clearance and organ uptake do occur. 111In-LiLo-IgM shows high relative accumulation in abscesses as well as high liver and spleen uptake. 111In-LiLo-IgM appears promising for imaging infection outside the trunk region.

  2. Cross-reactions in IgM ELISA tests to Legionella pneumophila sg1 and Bordetella pertussis among children suspected of legionellosis; potential impact of vaccination against pertussis?

    PubMed Central

    2015-01-01

    The objective of this study was preliminary evaluation of IgM cross-reaction in sera collected from children hospitalized because of suspected legionellosis. Sera with positive IgM results to L. pneumophila sgs1-7, B. pertussis or with simultaneous detection of IgM antibodies to L. pneumophila sgs1-7 and B. pertussis, or IgM to L. pneumophila sgs1-7 and M. pneumoniae in routine tests, were selected. In total, an adapted pre-absorption test was used for the serological confirmation of legionellosis in the sera of 19 children suspected of legionellosis, and also in 3 adult persons with confirmed Legionnaires’ disease. Sera were pre-absorbed with antigens of L. pneumophila sg1, B. pertussis or both, and tested by ELISA tests. The reduction of IgM antibody level by pre-absorption with antigen/antigens was determined. Reduction of anti-Lpsgs1-7 IgM by pre-absorption with L.pneumophila sg1 antigen ranged from 1.5 to 80, and reduction of anti-Bp IgM by pre-absorption with B. pertussis ranged from 2.0 to 23.8. Reduction by both antigens varied depending on the age of the patients: among children <4 yrs.old, the reduction of anti-B. pertussis IgM by both antigens was higher than for B. pertussis antigen alone. Based on the high difference (≥ 2 times) between reduction by L.pneumophila sg1 and by B. pertussis antigen, legionellosis was confirmed in 8/19 children. The majority of them also indicated IgM positive/borderline results for B. pertussis or M.pneumoniae in routine ELISA tests. As a preliminary, we posed a hypothesis of a potential impact of an anti-pertussis vaccination on the results obtained in anti-L. pneumophila ELISA IgM tests among young children. PMID:26557032

  3. An atypical IgM class platelet cold agglutinin induces GPVI-dependent aggregation of human platelets.

    PubMed

    Sánchez Guiu, I M; Martínez-Martinez, I; Martínez, C; Navarro-Fernandez, J; García-Candel, F; Ferrer-Marín, F; Vicente, V; Watson, S P; Andrews, R K; Gardiner, E E; Lozano, M L; Rivera, J

    2015-08-01

    Platelet cold agglutinins (PCA) cause pseudothrombocytopenia, spurious thrombocytopenia due to ex vivo platelet clumping, complicating clinical diagnosis, but mechanisms and consequences of PCA are not well defined. Here, we characterised an atypical immunoglobulin (Ig)M PCA in a 37-year-old woman with lifelong bleeding and chronic moderate thrombocytopenia, that induces activation and aggregation of autologous or allogeneic platelets via interaction with platelet glycoprotein (GP)VI. Patient temperature-dependent pseudothrombocytopenia was EDTA-independent, but was prevented by integrin αIIbβ3 blockade. Unstimulated patient platelets revealed elevated levels of bound IgM, increased expression of activation markers (P-selectin and CD63), low GPVI levels and abnormally high thromboxane (TX)A2 production. Patient serum induced temperature- and αIIbβ3-dependent decrease of platelet count in allogeneic donor citrated platelet-rich plasma (PRP), but not in PRP from Glanzmann's thrombasthenia or afibrinogenaemia patients. In allogeneic platelets, patient plasma induced shape change, P-selectin and CD63 expression, (14)C-serotonin release, and TXA2 production. Activation was not inhibited by aspirin, cangrelor or blocking anti-Fc receptor (FcγRIIA) antibody, but was abrogated by inhibitors of Src and Syk, and by a soluble GPVI-Fc fusion protein. GPVI-deficient platelets were not activated by patient plasma. These data provide the first evidence for an IgM PCA causing platelet activation/aggregation via GPVI. The PCA activity persisted over a five-year follow-up period, supporting a causative role in patient chronic thrombocytopenia and bleeding. PMID:25994029

  4. Cocoa and cocoa fibre differentially modulate IgA and IgM production at mucosal sites.

    PubMed

    Massot-Cladera, Malen; Franch, Àngels; Pérez-Cano, Francisco J; Castell, Margarida

    2016-05-01

    Previous studies have shown that a 10 % cocoa (C10) diet, containing polyphenols and fibre among others, modifies intestinal and systemic Ig production. The present study aimed at evaluating the impact of C10 on IgA and IgM production in the intestinal and extra-intestinal mucosal compartments, establishing the involvement of cocoa fibre (CF) in such effects. Mechanisms by which C10 intake may affect IgA synthesis in the salivary glands were also studied. To this effect, rats were fed either a standard diet, a diet containing C10, CF or inulin. Intestinal (the gut wash (GW), Peyer's patches (PP) and mesenteric lymph nodes (MLN)) and extra-intestinal (salivary glands) mucosal tissues and blood samples were collected for IgA and IgM quantification. The gene expressions of IgA production- and homing-related molecules were studied in the salivary glands. The C10 diet decreased intestinal IgA and IgM production. Although the CF diet decreased the GW IgA concentration, it increased PP, MLN and serum IgA concentrations. Both the C10 and the CF diets produced a down-regulatory effect on IgA secretion in the extra-intestinal tissues. The C10 diet interacted with the mechanisms involved in IgA synthesis, whereas the CF showed particular effects on the homing and transcytosis of IgA across the salivary glands. Overall, CF was able to up-regulate IgA production in the intestinal-inductor compartments, whereas it down-regulated its production at the mucosal-effector ones. Further studies must be directed to ascertain the mechanisms involved in the effect of particular cocoa components on gut-associated lymphoid tissue.

  5. Generation and Characterization of Antibodies against Asian Elephant (Elephas maximus) IgG, IgM, and IgA

    PubMed Central

    Humphreys, Alan F.; Tan, Jie; Peng, RongSheng; Benton, Susan M.; Qin, Xiang; Worley, Kim C.; Mikulski, Rose L.; Chow, Dar-Chone; Palzkill, Timothy G.; Ling, Paul D.

    2015-01-01

    Asian elephant (Elephas maximus) immunity is poorly characterized and understood. This gap in knowledge is particularly concerning as Asian elephants are an endangered species threatened by a newly discovered herpesvirus known as elephant endotheliotropic herpesvirus (EEHV), which is the leading cause of death for captive Asian elephants born after 1980 in North America. While reliable diagnostic assays have been developed to detect EEHV DNA, serological assays to evaluate elephant anti-EEHV antibody responses are lacking and will be needed for surveillance and epidemiological studies and also for evaluating potential treatments or vaccines against lethal EEHV infection. Previous studies have shown that Asian elephants produce IgG in serum, but they failed to detect IgM and IgA, further hampering development of informative serological assays for this species. To begin to address this issue, we determined the constant region genomic sequence of Asian elephant IgM and obtained some limited protein sequence information for putative serum IgA. The information was used to generate or identify specific commercial antisera reactive against IgM and IgA isotypes. In addition, we generated a monoclonal antibody against Asian elephant IgG. These three reagents were used to demonstrate that all three immunoglobulin isotypes are found in Asian elephant serum and milk and to detect antibody responses following tetanus toxoid booster vaccination or antibodies against a putative EEHV structural protein. The results indicate that these new reagents will be useful for developing sensitive and specific assays to detect and characterize elephant antibody responses for any pathogen or vaccine, including EEHV. PMID:25658336

  6. An atypical IgM class platelet cold agglutinin induces GPVI-dependent aggregation of human platelets.

    PubMed

    Sánchez Guiu, I M; Martínez-Martinez, I; Martínez, C; Navarro-Fernandez, J; García-Candel, F; Ferrer-Marín, F; Vicente, V; Watson, S P; Andrews, R K; Gardiner, E E; Lozano, M L; Rivera, J

    2015-08-01

    Platelet cold agglutinins (PCA) cause pseudothrombocytopenia, spurious thrombocytopenia due to ex vivo platelet clumping, complicating clinical diagnosis, but mechanisms and consequences of PCA are not well defined. Here, we characterised an atypical immunoglobulin (Ig)M PCA in a 37-year-old woman with lifelong bleeding and chronic moderate thrombocytopenia, that induces activation and aggregation of autologous or allogeneic platelets via interaction with platelet glycoprotein (GP)VI. Patient temperature-dependent pseudothrombocytopenia was EDTA-independent, but was prevented by integrin αIIbβ3 blockade. Unstimulated patient platelets revealed elevated levels of bound IgM, increased expression of activation markers (P-selectin and CD63), low GPVI levels and abnormally high thromboxane (TX)A2 production. Patient serum induced temperature- and αIIbβ3-dependent decrease of platelet count in allogeneic donor citrated platelet-rich plasma (PRP), but not in PRP from Glanzmann's thrombasthenia or afibrinogenaemia patients. In allogeneic platelets, patient plasma induced shape change, P-selectin and CD63 expression, (14)C-serotonin release, and TXA2 production. Activation was not inhibited by aspirin, cangrelor or blocking anti-Fc receptor (FcγRIIA) antibody, but was abrogated by inhibitors of Src and Syk, and by a soluble GPVI-Fc fusion protein. GPVI-deficient platelets were not activated by patient plasma. These data provide the first evidence for an IgM PCA causing platelet activation/aggregation via GPVI. The PCA activity persisted over a five-year follow-up period, supporting a causative role in patient chronic thrombocytopenia and bleeding.

  7. Multiplex Microsphere Immunoassays for the Detection of IgM and IgG to Arboviral Diseases

    PubMed Central

    Basile, Alison J.; Horiuchi, Kalanthe; Panella, Amanda J.; Laven, Janeen; Kosoy, Olga; Lanciotti, Robert S.; Venkateswaran, Neeraja; Biggerstaff, Brad J.

    2013-01-01

    Serodiagnosis of arthropod-borne viruses (arboviruses) at the Division of Vector-Borne Diseases, CDC, employs a combination of individual enzyme-linked immunosorbent assays and microsphere immunoassays (MIAs) to test for IgM and IgG, followed by confirmatory plaque-reduction neutralization tests. Based upon the geographic origin of a sample, it may be tested concurrently for multiple arboviruses, which can be a cumbersome task. The advent of multiplexing represents an opportunity to streamline these types of assays; however, because serologic cross-reactivity of the arboviral antigens often confounds results, it is of interest to employ data analysis methods that address this issue. Here, we constructed 13-virus multiplexed IgM and IgG MIAs that included internal and external controls, based upon the Luminex platform. Results from samples tested using these methods were analyzed using 8 different statistical schemes to identify the best way to classify the data. Geographic batteries were also devised to serve as a more practical diagnostic format, and further samples were tested using the abbreviated multiplexes. Comparative error rates for the classification schemes identified a specific boosting method based on logistic regression “Logitboost” as the classification method of choice. When the data from all samples tested were combined into one set, error rates from the multiplex IgM and IgG MIAs were <5% for all geographic batteries. This work represents both the most comprehensive, validated multiplexing method for arboviruses to date, and also the most systematic attempt to determine the most useful classification method for use with these types of serologic tests. PMID:24086608

  8. Cocoa and cocoa fibre differentially modulate IgA and IgM production at mucosal sites.

    PubMed

    Massot-Cladera, Malen; Franch, Àngels; Pérez-Cano, Francisco J; Castell, Margarida

    2016-05-01

    Previous studies have shown that a 10 % cocoa (C10) diet, containing polyphenols and fibre among others, modifies intestinal and systemic Ig production. The present study aimed at evaluating the impact of C10 on IgA and IgM production in the intestinal and extra-intestinal mucosal compartments, establishing the involvement of cocoa fibre (CF) in such effects. Mechanisms by which C10 intake may affect IgA synthesis in the salivary glands were also studied. To this effect, rats were fed either a standard diet, a diet containing C10, CF or inulin. Intestinal (the gut wash (GW), Peyer's patches (PP) and mesenteric lymph nodes (MLN)) and extra-intestinal (salivary glands) mucosal tissues and blood samples were collected for IgA and IgM quantification. The gene expressions of IgA production- and homing-related molecules were studied in the salivary glands. The C10 diet decreased intestinal IgA and IgM production. Although the CF diet decreased the GW IgA concentration, it increased PP, MLN and serum IgA concentrations. Both the C10 and the CF diets produced a down-regulatory effect on IgA secretion in the extra-intestinal tissues. The C10 diet interacted with the mechanisms involved in IgA synthesis, whereas the CF showed particular effects on the homing and transcytosis of IgA across the salivary glands. Overall, CF was able to up-regulate IgA production in the intestinal-inductor compartments, whereas it down-regulated its production at the mucosal-effector ones. Further studies must be directed to ascertain the mechanisms involved in the effect of particular cocoa components on gut-associated lymphoid tissue. PMID:26975903

  9. Mapping the low-surface brightness Universe in the UV band with Lyα emission from IGM filaments

    NASA Astrophysics Data System (ADS)

    Silva, Marta B.; Kooistra, Robin; Zaroubi, Saleem

    2016-10-01

    A large fraction of the baryonic matter in the Universe is located in filaments in the intergalactic medium (IGM). However, the low surface brightness of these filaments has not yet allowed their direct detection except in very special regions in the circum-galactic medium. Here we simulate the intensity and spatial fluctuations in Lyman alpha (Lyα) emission from filaments in the IGM and discuss the prospects for the next generation of space-based instruments to detect the low-surface brightness Universe at ultraviolet (UV) wavelengths. Starting with a high-resolution N-body simulation, we obtain the dark matter density fluctuations and associate baryons with the dark matter particles assuming that they follow the same spatial distribution. The IGM thermal and ionization state is set by a model of the UV background and by the relevant cooling processes for a hydrogen and helium gas. The Lyα emissivity is then estimated, taking into account recombination and collisional excitation processes. We find that the detection of these filaments through their Lyα emission is well in the reach of the next generation of UV space-based instruments and so it should be achieved in the next decade. The density field is populated with halos and galaxies and their Lyα emission is estimated. Galaxies are treated as foregrounds and so we discuss methods to reduce their contamination from observational maps. Finally, we estimate the UV continuum background as a function of the redshift of the Lyα emission line and discuss how this continuum can affect observations.

  10. Generation and characterization of antibodies against Asian elephant (Elephas maximus) IgG, IgM, and IgA.

    PubMed

    Humphreys, Alan F; Tan, Jie; Peng, RongSheng; Benton, Susan M; Qin, Xiang; Worley, Kim C; Mikulski, Rose L; Chow, Dar-Chone; Palzkill, Timothy G; Ling, Paul D

    2015-01-01

    Asian elephant (Elephas maximus) immunity is poorly characterized and understood. This gap in knowledge is particularly concerning as Asian elephants are an endangered species threatened by a newly discovered herpesvirus known as elephant endotheliotropic herpesvirus (EEHV), which is the leading cause of death for captive Asian elephants born after 1980 in North America. While reliable diagnostic assays have been developed to detect EEHV DNA, serological assays to evaluate elephant anti-EEHV antibody responses are lacking and will be needed for surveillance and epidemiological studies and also for evaluating potential treatments or vaccines against lethal EEHV infection. Previous studies have shown that Asian elephants produce IgG in serum, but they failed to detect IgM and IgA, further hampering development of informative serological assays for this species. To begin to address this issue, we determined the constant region genomic sequence of Asian elephant IgM and obtained some limited protein sequence information for putative serum IgA. The information was used to generate or identify specific commercial antisera reactive against IgM and IgA isotypes. In addition, we generated a monoclonal antibody against Asian elephant IgG. These three reagents were used to demonstrate that all three immunoglobulin isotypes are found in Asian elephant serum and milk and to detect antibody responses following tetanus toxoid booster vaccination or antibodies against a putative EEHV structural protein. The results indicate that these new reagents will be useful for developing sensitive and specific assays to detect and characterize elephant antibody responses for any pathogen or vaccine, including EEHV.

  11. Generation and characterization of antibodies against Asian elephant (Elephas maximus) IgG, IgM, and IgA.

    PubMed

    Humphreys, Alan F; Tan, Jie; Peng, RongSheng; Benton, Susan M; Qin, Xiang; Worley, Kim C; Mikulski, Rose L; Chow, Dar-Chone; Palzkill, Timothy G; Ling, Paul D

    2015-01-01

    Asian elephant (Elephas maximus) immunity is poorly characterized and understood. This gap in knowledge is particularly concerning as Asian elephants are an endangered species threatened by a newly discovered herpesvirus known as elephant endotheliotropic herpesvirus (EEHV), which is the leading cause of death for captive Asian elephants born after 1980 in North America. While reliable diagnostic assays have been developed to detect EEHV DNA, serological assays to evaluate elephant anti-EEHV antibody responses are lacking and will be needed for surveillance and epidemiological studies and also for evaluating potential treatments or vaccines against lethal EEHV infection. Previous studies have shown that Asian elephants produce IgG in serum, but they failed to detect IgM and IgA, further hampering development of informative serological assays for this species. To begin to address this issue, we determined the constant region genomic sequence of Asian elephant IgM and obtained some limited protein sequence information for putative serum IgA. The information was used to generate or identify specific commercial antisera reactive against IgM and IgA isotypes. In addition, we generated a monoclonal antibody against Asian elephant IgG. These three reagents were used to demonstrate that all three immunoglobulin isotypes are found in Asian elephant serum and milk and to detect antibody responses following tetanus toxoid booster vaccination or antibodies against a putative EEHV structural protein. The results indicate that these new reagents will be useful for developing sensitive and specific assays to detect and characterize elephant antibody responses for any pathogen or vaccine, including EEHV. PMID:25658336

  12. IGM CONSTRAINTS FROM THE SDSS-III/BOSS DR9 Lyα FOREST TRANSMISSION PROBABILITY DISTRIBUTION FUNCTION

    SciTech Connect

    Lee, Khee-Gan; Hennawi, Joseph F.; Spergel, David N.; Weinberg, David H.; Hogg, David W.; Viel, Matteo; Bolton, James S.; Bailey, Stephen; Carithers, William; Schlegel, David J.; Pieri, Matthew M.; Lundgren, Britt; Palanque-Delabrouille, Nathalie; Yèche, Christophe; Schneider, Donald P.

    2015-02-01

    The Lyα forest transmission probability distribution function (PDF) is an established probe of the intergalactic medium (IGM) astrophysics, especially the temperature-density relationship of the IGM. We measure the transmission PDF from 3393 Baryon Oscillations Spectroscopic Survey (BOSS) quasars from Sloan Digital Sky Survey Data Release 9, and compare with mock spectra that include careful modeling of the noise, continuum, and astrophysical uncertainties. The BOSS transmission PDFs, measured at (z) = [2.3, 2.6, 3.0], are compared with PDFs created from mock spectra drawn from a suite of hydrodynamical simulations that sample the IGM temperature-density relationship, γ, and temperature at mean density, T {sub 0}, where T(Δ) = T {sub 0}Δ{sup γ} {sup –} {sup 1}. We find that a significant population of partial Lyman-limit systems (LLSs) with a column-density distribution slope of β{sub pLLS} ∼ – 2 are required to explain the data at the low-transmission end of transmission PDF, while uncertainties in the mean Lyα forest transmission affect the high-transmission end. After modeling the LLSs and marginalizing over mean transmission uncertainties, we find that γ = 1.6 best describes the data over our entire redshift range, although constraints on T {sub 0} are affected by systematic uncertainties. Within our model framework, isothermal or inverted temperature-density relationships (γ ≤ 1) are disfavored at a significance of over 4σ, although this could be somewhat weakened by cosmological and astrophysical uncertainties that we did not model.

  13. Report of a joint DMRQC/Organon field trial to detect hepatitis A IgM by ELISA.

    PubMed

    Supran, E M; Craske, J; Hart, R J; Kurtz, J B; Parry, J V; Skidmore, S J; Gardner, P S

    1983-10-01

    The results of a field trial of a joint DMRQC/Organon ELISA kit for the detection of hepatitis A IgM antibody are reported. The participating laboratories were asked to use the kit to test a panel of 360 specimens consisting of duplicate coded samples of 180 sera. The panel was also tested by MACRIA in the Virus Reference Laboratory, Colindale. The ELISA was shown to be specific and sensitive giving good discrimination between acute and late convalescent hepatitis A sera. It was proposed that the same cut-off control as is used in the RIA (equivalent to 10 RIA units) should be adopted for the ELISA also.

  14. Genetic Resistance to Malaria Is Associated With Greater Enhancement of Immunoglobulin (Ig)M Than IgG Responses to a Broad Array of Plasmodium falciparum Antigens

    PubMed Central

    Arama, Charles; Skinner, Jeff; Doumtabe, Didier; Portugal, Silvia; Tran, Tuan M.; Jain, Aarti; Traore, Boubacar; Doumbo, Ogobara K.; Davies, David Huw; Troye-Blomberg, Marita; Dolo, Amagana; Felgner, Philip L.; Crompton, Peter D.

    2015-01-01

    Background. People of the Fulani ethnic group are more resistant to malaria compared with genetically distinct ethnic groups, such as the Dogon people, in West Africa, and studies suggest that this resistance is mediated by enhanced antibody responses to Plasmodium falciparum antigens. However, prior studies measured antibody responses to <0.1% of P falciparum proteins, so whether the Fulani mount an enhanced and broadly reactive immunoglobulin (Ig)M and IgG response to P falciparum remains unknown. In general, little is known about the extent to which host genetics influence the overall antigen specificity of IgM and IgG responses to natural infections. Methods. In a cross-sectional study in Mali, we collected plasma from asymptomatic, age-matched Fulani (n = 24) and Dogon (n = 22) adults with or without concurrent P falciparum infection. We probed plasma against a protein microarray containing 1087 P falciparum antigens and compared IgM and IgG profiles by ethnicity. Results. We found that the breadth and magnitude of P falciparum-specific IgM and IgG responses were significantly higher in the malaria-resistant Fulani versus the malaria-susceptible Dogon, and, unexpectedly, P falciparum-specific IgM responses more strongly distinguished the 2 ethnic groups. Conclusions. These findings point to an underappreciated role for IgM in protection from malaria, and they suggest that host genetics may influence the antigen specificity of IgM and IgG responses to infection. PMID:26361633

  15. Rapid Proliferation and Differentiation of a Subset of Circulating IgM Memory B Cells to a CpG/Cytokine Stimulus In Vitro.

    PubMed

    Vásquez, Camilo; Franco, Manuel A; Angel, Juana

    2015-01-01

    Circulating human IgM expressing memory B cells have been incompletely characterized. Here, we compared the phenotype and in vitro functional response (capacity to proliferate and differentiate to antibody secreting cells) in response to CpG and a cytokine cocktail (IL-2, IL-6, and IL-10) of sorted naïve B cells, IgM memory B cells and isotype-switched circulating memory B cells. Compared to naïve B cells, IgM memory B cells had lower integrated mean fluorescence intensity (iMFI) of BAFF-R, CD38, CD73, and IL-21R, but higher iMFI of CD95, CD11c, TLR9, PD-1, and CD122. Compared to switched memory B cells, IgM memory B cells had higher iMFI of BAFF-R, PD-1, IL-21R, TLR9, and CD122, but lower iMFI of CD38, CD95, and CD73. Four days after receiving the CpG/cytokine cocktail, higher frequencies of IgM than switched memory B cells-and these in turn greater than naïve cells-proliferated and differentiated to antibody secreting cells. At this time point, a small percentage (median of 7.6%) of stimulated IgM memory B cells changed isotype to IgG. Thus, among the heterogeneous population of human circulating IgM memory B cells a subset is capable of a rapid functional response to a CpG/cytokine stimulus in vitro.

  16. Serum carbohydrate-binding IgM are present in Vietnamese striped catfish (Pangasianodon hypophthalmus) but not in North African catfish (Clarias gariepinus).

    PubMed

    Giang, Duong Thi Huong; Van Driessche, Edilbert; Beeckmans, Sonia

    2012-02-01

    Pangasianodon hypophthalmus serum was fractionated by affinity chromatography on 12 different Sepharose-carbohydrate columns and proteins eluted by the corresponding sugar. Binding to the affinity matrices is dependent on Ca(2+) ions. Upon gel filtration using Superose-12, essentially one fraction was obtained, eluting as a protein with a molecular mass of about 900 kDa. SDS-PAGE in reducing conditions revealed the presence of large (72 kDa) subunits (H-chains) and one up to three small (24, 26 and/or 28-29 kDa) subunits (L-chains). The isolated proteins were shown to be IgM since they bind monoclonal anti-P. hypophthalmus IgM antibodies. Rabbit polyclonal anti-galactose-binding IgM only cross-react with some sugar-binding IgM. The H-chains of the anti-carbohydrate IgM are glycosylated. Circular dichroism studies revealed that the IgMs have an "all-β" type of structure, and that Ca(2+) ions, though essential for carbohydrate-binding activity, are not required for the structural integrity of the molecules. In non-reducing SDS-PAGE, only monomers and halfmers were obtained, showing that there are no disulfide bonds linking the monomers, and that a disulfide bond connecting both H-chains within one monomer is only present in 45% of the molecules. Both the monomers and the halfmers display molecular mass heterogeneity which is indicative for redox forms at the level of the intradomain disulfide bonds. The native carbohydrate-binding IgMs agglutinate erythrocytes from different animals, as well as fish pathogenic bacteria. Similar proteins could not be isolated from another catfish, Clarias gariepinus. PMID:21911003

  17. Decreased frequency and activated phenotype of blood CD27 IgD IgM B lymphocytes is a permanent abnormality in systemic lupus erythematosus patients

    PubMed Central

    2010-01-01

    Introduction Systemic lupus erythematosus (SLE) is characterized by B cell hyper-activation and auto-reactivity resulting in pathogenic auto-antibody generation. The phenotypic analysis of blood B cell subsets can be used to understand these alterations. Methods The combined detection of CD19, CD27 and IgD (or IgM) by flow cytometry (FC) analysis delineates five well-defined blood B cell-subsets: naive, switched (S) memory, double negative (DN) memory and CD27 IgD IgM (non-switched memory) B lymphocytes, and plasma cells (PCs). This phenotypic study was performed in 69 consecutive SLE patients and 31 healthy controls. Results SLE patients exhibited several abnormalities in the distribution of these B cell subsets, including elevated levels of DN memory B cells and PCs, and decreased CD27 IgD IgM B cells. Active SLE patients also showed decreased presence of S memory B cells and increased proportions of naive B lymphocytes. Nevertheless, when the patients in remission who did not require treatment were studied separately, the only remaining abnormality was a reduction of the CD27 IgD IgM B cell-subset detectable in most of these patients. The level of reduction of CD27 IgD IgM B cells was associated with elevated values of serum SLE auto-antibodies. Further analysis of this latter B cell-subset specifically showed increased expression of CD80, CD86, CD95, 9G4 idiotype and functional CXCR3 and CXCR4. Conclusions The presence of a reduced blood CD27 IgD IgM B cell-subset, exhibiting an activated state and enriched for auto-reactivity, is a consistent B cell abnormality in SLE. These findings suggest that CD27 IgD IgM B lymphocytes play a role in the pathogenesis of this disease. PMID:20525218

  18. Modulation of natural IgM autoantibodies to oxidative stress-related neo-epitopes on apoptotic cells in newborns of mothers with anti-Ro autoimmunity.

    PubMed

    Grönwall, Caroline; Clancy, Robert M; Getu, Lelise; Lloyd, Katy A; Siegel, Don L; Reed, Joanne H; Buyon, Jill P; Silverman, Gregg J

    2016-09-01

    At birth, the human immune system already contains substantial levels of polymeric IgM, that include autoantibodies to neo-epitopes on apoptotic cells (ACs) that are proposed to play homeostatic and anti-inflammatory roles. Yet the biologic origins and developmental regulation of these naturally arising antibodies remain poorly understood. Herein, we report that levels of IgM-antibodies to malondialdehyde (MDA) protein adducts, a common type of in vivo generated oxidative stress-related neoepitope, directly correlate with the relative binding of neonatal-IgM to ACs. Levels of IgM to phosphorylcholine (PC), a natural antibody prevalent in adults, were relatively scant in cord blood, while there was significantly greater relative representation of IgM anti-MDA antibodies in newborns compared to adults. To investigate the potential interrelationships between neonatal IgM with pathogenic IgG-autoantibodies, we studied 103 newborns born to autoimmune mothers with IgG anti-Ro (i.e., 70 with neonatal lupus and 33 without neonatal lupus). In these subjects the mean levels of IgM anti-Ro60 were significantly higher than in the newborns from non-autoimmune mothers. In contrast, levels of IgM anti-MDA in IgG anti-Ro exposed neonates were significantly lower than in neonates from non-autoimmune mothers. The presence or absence of neonatal lupus did not appear to influence the total levels of IgM in the anti-Ro exposed newborns. Taken together, our studies provide evidence that the immune development of the natural IgM-repertoire may be affected, and become imprinted by, the transfer of maternal IgG into the fetus. PMID:27289167

  19. Serum IgM antibodies contribute to high levels of opsonophagocytic activities in toddlers immunized with a single dose of the 9-valent pneumococcal conjugate vaccine.

    PubMed

    Simell, Birgit; Nurkka, Anu; Ekström, Nina; Givon-Lavi, Noga; Käyhty, Helena; Dagan, Ron

    2012-10-01

    In immunogenicity trials of pneumococcal conjugate vaccines (PCVs), only IgG antibody concentrations to pneumococcal capsular polysaccharides (PPSs) are usually determined, along with the opsonophagocytic activity (OPA) of antipneumococcal antibodies. We aimed to determine the role of both IgG and IgM in OPA in toddlers receiving one dose of 9-valent PCV (PCV9). The IgG and IgM antibody concentrations to PPSs of serotypes 6A, 9V, 14, 19F, and 23F were measured by enzyme immunoassay in sera from toddlers (ages 18 to 35 months) 1 month after a single PCV9 dose. The OPA for the same serotypes was measured by multiplexed opsonophagocytosis assay (MOPA). Further, IgG and IgM concentrations and MOPA were measured to PPS of serotypes 6A, 14, and 19F in sera collected 12 months after vaccination. The detected MOPA titers were high in comparison to the IgG concentrations 1 month after immunization. The IgM concentrations were higher than IgG concentrations for serotypes 6A and 14 (P < 0.001) and as high as IgG for serotypes 9V, 19F, and 23F. Correlation of the IgM antibody concentrations with MOPA (r = 0.35 to 0.65) was stronger compared to that of the IgG antibodies (r = 0.07 to 0.41). The depletion of IgG antibodies in three sets of pooled sera only slightly decreased the OPA activity against serotype 14. At 12 months after immunization, 50 to 100% of serum samples still showed detectable MOPA activity against serotypes 6A, 14, and 19F. Our results suggest that IgM contributes to OPA 1 month after a single PCV9 vaccination in toddlers and that functionally active IgM and IgG antibodies persist for at least a year. PMID:22875604

  20. Modulation of natural IgM autoantibodies to oxidative stress-related neo-epitopes on apoptotic cells in newborns of mothers with anti-Ro autoimmunity.

    PubMed

    Grönwall, Caroline; Clancy, Robert M; Getu, Lelise; Lloyd, Katy A; Siegel, Don L; Reed, Joanne H; Buyon, Jill P; Silverman, Gregg J

    2016-09-01

    At birth, the human immune system already contains substantial levels of polymeric IgM, that include autoantibodies to neo-epitopes on apoptotic cells (ACs) that are proposed to play homeostatic and anti-inflammatory roles. Yet the biologic origins and developmental regulation of these naturally arising antibodies remain poorly understood. Herein, we report that levels of IgM-antibodies to malondialdehyde (MDA) protein adducts, a common type of in vivo generated oxidative stress-related neoepitope, directly correlate with the relative binding of neonatal-IgM to ACs. Levels of IgM to phosphorylcholine (PC), a natural antibody prevalent in adults, were relatively scant in cord blood, while there was significantly greater relative representation of IgM anti-MDA antibodies in newborns compared to adults. To investigate the potential interrelationships between neonatal IgM with pathogenic IgG-autoantibodies, we studied 103 newborns born to autoimmune mothers with IgG anti-Ro (i.e., 70 with neonatal lupus and 33 without neonatal lupus). In these subjects the mean levels of IgM anti-Ro60 were significantly higher than in the newborns from non-autoimmune mothers. In contrast, levels of IgM anti-MDA in IgG anti-Ro exposed neonates were significantly lower than in neonates from non-autoimmune mothers. The presence or absence of neonatal lupus did not appear to influence the total levels of IgM in the anti-Ro exposed newborns. Taken together, our studies provide evidence that the immune development of the natural IgM-repertoire may be affected, and become imprinted by, the transfer of maternal IgG into the fetus.

  1. Simultaneous expression of Borrelia OspA and OspC and IgM response in cerebrospinal fluid in early neurologic Lyme disease.

    PubMed Central

    Schutzer, S E; Coyle, P K; Krupp, L B; Deng, Z; Belman, A L; Dattwyler, R; Luft, B J

    1997-01-01

    Lyme disease is the major tick-borne disease, caused by Borrelia burgdorferi (Bb). Neurological involvement is common in all stages. In vivo expression of Bb antigens (Ags) and the immune response to them has not been well investigated in the cerebrospinal fluid (CSF). Upregulation of outer surface protein (Osp) C and concomitant downregulation of OspA before tick inoculation of the spirochete has been reported in skin and blood in animals. CSF OspA Ag in early disease suggests otherwise in CSF. Early Ag expression and IgM response in human CSF was investigated here. Paired CSF and serum was collected from 16 early, predominantly erythema migrans Lyme disease patients with neurologic problems, 13 late Lyme disease patients, and 19 other neurologic disease (OND) controls. Samples were examined for IgM reactivity to recombinant Bb-specific Osps using ELISA and immunoblot. Of 12 early Lyme disease patients with neurologic involvement with both CSF and serum IgM against OspC, 7 (58%) had IgM to OspA (n = 5) or OspB (n = 2) that was restricted to the CSF, not serum. Overall, 12 of 16 (75%) of these early Lyme disease patients with neurologic involvement had CSF and serum IgM against OspC. Only 3 of 13 (23%) late Lyme disease patients and none of 19 OND controls had CSF IgM directed against OspC. In conclusion, in CSF, OspC and OspA can be coexpressed, and IgM response to them occurs in early Lyme disease patients with neurologic involvement. This biologic finding may also provide a discriminating marker for CNS infection in Lyme disease. PMID:9259573

  2. ALS - resources

    MedlinePlus

    Resources - ALS ... The following organizations are good resources for information on amyotrophic lateral sclerosis : Muscular Dystrophy Association -- mda.org/disease/amyotrophic-lateral-sclerosis National Amyotrophic Lateral Sclerosis (ALS) Registry -- ...

  3. Detection of IgA and IgM antibodies to HIV-1 in neonates by radioimmune western blotting.

    PubMed Central

    Portincasa, P.; Conti, G.; Re, M. C.; Chezzi, C.

    1992-01-01

    OBJECTIVE--To detect infection with HIV-1 by IgA and IgM response at birth in children born to HIV-1 seropositive mothers. DESIGN--Western blotting and radioimmune western blotting on stored sera from infected and uninfected babies born to HIV-1 seropositive mothers. Sera were pretreated to remove IgG. SETTING--Parma and Bologna, Italy. SUBJECTS--12 infected and five uninfected babies born to HIV-1 seropositive mothers and three babies born to seronegative mothers. MAIN OUTCOME MEASURES--Effectiveness of western blotting and radioimmune western blotting in detecting antibodies to HIV-1 gene products. RESULTS--With conventional western blotting we found IgA class antibodies to HIV-1 proteins in serum from three out of 12 infected children; in two of these three the serum was collected at age 3 months (positive controls). Radioimmune western blotting detected both IgA and IgM antibodies in serum from all infected children tested, whereas all serum from uninfected children born to seropositive and seronegative mothers showed no such antibodies. CONCLUSION--Although the technique should be tested on more patients, radioimmune western blotting seems to be a valuable tool for serological diagnosis of congenital HIV-1 infection at birth in neonates born to seropositive mothers. Images FIG 1 FIG 2 PMID:1628052

  4. Inflammation rapidly reorganizes mouse bone marrow B cells and their environment in conjunction with early IgM responses.

    PubMed

    Moreau, Joshua M; Berger, Alexandra; Nelles, Megan E; Mielnik, Michael; Furlonger, Caren; Cen, Selena Y; Besla, Rickvinder; Robbins, Clinton S; Paige, Christopher J

    2015-09-01

    Systemic inflammation perturbs the bone marrow environment by evicting resident B cells and favoring granulopoiesis over lymphopoiesis. Despite these conditions, a subset of marrow B cell remains to become activated and produce potent acute immunoglobulin M (IgM) responses. This discrepancy is currently unresolved and a complete characterization of early perturbations in the B-cell niche has not been undertaken. Here, we show that within a few hours of challenging mice with adjuvant or cecal puncture, B cells accumulate in the bone marrow redistributed away from sinusoid vessels. This response correlates with enhanced sensitivity to CXC chemokine ligand 12 (CXCL12) but not CXCL13 or CC chemokine ligand 21. Concurrently, a number of B-cell survival and differentiation factors are elevated to produce a transiently supportive milieu. Disrupting homing dynamics with a CXC chemokine receptor 4 inhibitor reduced the formation of IgM-secreting cells. These data highlight the rapidity with which peripheral inflammation modifies the marrow compartment, and demonstrate that such modifications regulate acute IgM production within this organ. Furthermore, our study indicates that conversion to a state of emergency granulopoiesis is temporally delayed, allowing B cells opportunity to respond to antigen.

  5. [The confusing diversity of IgM tests in the diagnosis of Toxoplasma infections: efforts towards an optimal strategy].

    PubMed

    Auer, Herbert; Vander-Möse, Angelika; Aspöck, Horst

    2003-01-01

    IgM antibodies are indicative for a recent infection, thus the detection of this isotype is of essential significance particularly in the diagnosis of infections with Toxoplasma gondii during pregnancy (primary infection, seroconversion). Numerous serological tests and test kits (e.g. indirect immunofluorescent assay/IFAT, enzyme-linked immunosorbent assay/ELISA, Immunosorbent agglutination assay/ISAGA, Westernblot/WB) using different antigens and antigen preparations are provided by numerous companies. The sensitivities of such tests, however, variy considerably: The serological test results of four pregnant women with seroconversions and of three newborns from mothers with seroconversions are presented: VIDAS M and ISAGA M from one company yielded false negative results in three pregnant women whereas ISAGA M from another company could detect specific IgM. However, examination of the cord blood of the three newborns unanimously revealed IgM-negative results. Thus, our diagnostic strategy for pregnant women includes IIFT (or SFT) as basic test and ISAGA M (Toxotool I from Innogenetics) as well as IgG avidity test as additional tests; the serological diagnosis of suspected congenital infection comprises IFAT (or SFT), ISAGA M (from Innogenetics) and IgM/IgG Westernblot.

  6. Fat-associated lymphoid clusters control local IgM secretion during pleural infection and lung inflammation.

    PubMed

    Jackson-Jones, Lucy H; Duncan, Sheelagh M; Magalhaes, Marlène S; Campbell, Sharon M; Maizels, Rick M; McSorley, Henry J; Allen, Judith E; Bénézech, Cécile

    2016-01-01

    Fat-associated lymphoid clusters (FALC) are inducible structures that support rapid innate-like B-cell immune responses in the serous cavities. Little is known about the physiological cues that activate FALCs in the pleural cavity and more generally the mechanisms controlling B-cell activation in FALCs. Here we show, using separate models of pleural nematode infection with Litomosoides sigmodontis and Altenaria alternata induced acute lung inflammation, that inflammation of the pleural cavity rapidly activates mediastinal and pericardial FALCs. IL-33 produced by FALC stroma is crucial for pleural B1-cell activation and local IgM secretion. However, B1 cells are not the direct target of IL-33, which instead requires IL-5 for activation. Moreover, lung inflammation leads to increased IL-5 production by type 2 cytokine-producing innate lymphoid cells (ILC2) in the FALC. These findings reveal a link between inflammation, IL-33 release by FALC stromal cells, ILC2 activation and pleural B-cell activation in FALCs, resulting in local and antigen-specific IgM production. PMID:27582256

  7. Changes of tetanus specific IgG, IgM and IgG subclasses after DPT vaccination.

    PubMed

    Kim, J S; Kim, S J; Shin, K J; Hwang, P H; Cho, S C

    1989-01-01

    We evaluated tetanus specific IgG, IgM, IgG subclasses after DPT vaccination in infants and children. Tetanus toxoid specific IgG, IgM IgG subclasses were measured to characterize the isotope profile of antibody against tetanus toxoid. The values of the tetanus specific IgG in the positive group were significantly increased compared to those of the control group, and were significantly increased after two inoculation. Tetanus specific IgG was very low in adults and neonates. In our tetanus specific IgG subclasses study, forty-five of 56 cases (80%) showed predominantly IgG1 antibody responses to tetanus toxoid, while twenty-five of 56 cases (45%) showed IgG4 responses. Both IgG1 and IgG4 responses were demonstrated in 17 cases (30%). So we suggest that IgG was mainly involved in humoral immune response after DPT vaccination, and IgG1 may play an important role among IgG subclasses. IgG4, alone or together with IgG1, can also play a role in immune response to tetanus toxoid.

  8. Fat-associated lymphoid clusters control local IgM secretion during pleural infection and lung inflammation

    PubMed Central

    Jackson-Jones, Lucy H.; Duncan, Sheelagh M.; Magalhaes, Marlène S.; Campbell, Sharon M.; Maizels, Rick M.; McSorley, Henry J.; Allen, Judith E.; Bénézech, Cécile

    2016-01-01

    Fat-associated lymphoid clusters (FALC) are inducible structures that support rapid innate-like B-cell immune responses in the serous cavities. Little is known about the physiological cues that activate FALCs in the pleural cavity and more generally the mechanisms controlling B-cell activation in FALCs. Here we show, using separate models of pleural nematode infection with Litomosoides sigmodontis and Altenaria alternata induced acute lung inflammation, that inflammation of the pleural cavity rapidly activates mediastinal and pericardial FALCs. IL-33 produced by FALC stroma is crucial for pleural B1-cell activation and local IgM secretion. However, B1 cells are not the direct target of IL-33, which instead requires IL-5 for activation. Moreover, lung inflammation leads to increased IL-5 production by type 2 cytokine-producing innate lymphoid cells (ILC2) in the FALC. These findings reveal a link between inflammation, IL-33 release by FALC stromal cells, ILC2 activation and pleural B-cell activation in FALCs, resulting in local and antigen-specific IgM production. PMID:27582256

  9. Differential diagnosis of IgM MGUS and WM according to B-lymphoid infiltration by morphology and flow cytometry.

    PubMed

    Ocio, Enrique M; del Carpio, Daniel; Caballero, Álvaro; Alonso, Jennifer; Paiva, Bruno; Pesoa, Roberto; Villaescusa, Teresa; López-Anglada, Lucía; Vidriales, Belén; García-Sanz, Ramón

    2011-02-01

    The distinction between IgM monoclonal gammopathy of undetermined significance (MGUS), asymptomatic Waldenstrom's macroglobulinemia (WM; aWM), and symptomatic WM (sWM) relies on two features: the presence of infiltration by lymphoplasmacytic lymphoma in the bone marrow (BM) biopsy and the existence of signs or symptoms attributable to the disease. Nevertheless, some patients lack a BM biopsy or it is not conclusive for diagnosis. In this study we have investigated 94 patients with IgM monoclonal gammopathies, in which a BM trephine biopsy and morphological and flow cytometry (FCM) evaluation of BM aspirate were available at diagnosis. We found a clear correlation between BM infiltration of B-lymphocytes assessed by morphology and by FCM with a Pearson correlation of 0.62 (P<.001). Moreover, in the absence of a BM trephine biopsy, the cut-off points that would help in the differential diagnosis between MGUS, aWM, and sWM would be 20% for morphology and 5% for FCM, both assessed in the BM aspirate. PMID:21454201

  10. Diagnostic accuracy of an IgM enzyme-linked immunosorbent assay and comparison with 2 polymerase chain reactions for early diagnosis of human leptospirosis.

    PubMed

    Vanasco, N B; Jacob, P; Landolt, N; Chiani, Y; Schmeling, M F; Cudos, C; Tarabla, H; Lottersberger, J

    2016-04-01

    Enzyme-linked immunosorbent assay (ELISA) tests and polymerase chain reaction (PCR) may play a key role for early detection and treatment of human leptospirosis in developing countries. The aims of this study were to develop and validate an IgM ELISA under field conditions and to compare the diagnostic accuracy among IgG, IgM ELISAs, conventional PCR (cPCR), and real-time PCR (rtPCR) for early detection of human leptospirosis. Overall accuracy of IgM ELISA was sensitivity of 87.9%, specificity of 97.0%, and area under the curve of 0.940. When the 4 methods were compared, IgM ELISA showed the greatest diagnostic accuracy (J=0.6) followed by rtPCR (J=0.4), cPCR (J=0.2) and IgG ELISA (J=0.1). Our results support the use of IgM ELISA and rtPCR for early diagnosis of the disease. Moreover, due to their high specificity, they could be also useful to replace or supplement microscopic agglutination test as a confirmatory test, allowing more confirmations.

  11. Diagnostic accuracy of an IgM enzyme-linked immunosorbent assay and comparison with 2 polymerase chain reactions for early diagnosis of human leptospirosis.

    PubMed

    Vanasco, N B; Jacob, P; Landolt, N; Chiani, Y; Schmeling, M F; Cudos, C; Tarabla, H; Lottersberger, J

    2016-04-01

    Enzyme-linked immunosorbent assay (ELISA) tests and polymerase chain reaction (PCR) may play a key role for early detection and treatment of human leptospirosis in developing countries. The aims of this study were to develop and validate an IgM ELISA under field conditions and to compare the diagnostic accuracy among IgG, IgM ELISAs, conventional PCR (cPCR), and real-time PCR (rtPCR) for early detection of human leptospirosis. Overall accuracy of IgM ELISA was sensitivity of 87.9%, specificity of 97.0%, and area under the curve of 0.940. When the 4 methods were compared, IgM ELISA showed the greatest diagnostic accuracy (J=0.6) followed by rtPCR (J=0.4), cPCR (J=0.2) and IgG ELISA (J=0.1). Our results support the use of IgM ELISA and rtPCR for early diagnosis of the disease. Moreover, due to their high specificity, they could be also useful to replace or supplement microscopic agglutination test as a confirmatory test, allowing more confirmations. PMID:26867967

  12. Functional properties of isotype-switched immunoglobulin M (IgM) and IgG monoclonal antibodies to Pseudomonas aeruginosa lipopolysaccharide.

    PubMed Central

    Pollack, M; Koles, N L; Preston, M J; Brown, B J; Pier, G B

    1995-01-01

    Controversy exists regarding isotype-related differences in the antibacterial and protective properties of lipopolysaccharide (LPS)-specific antibodies of the immunoglobulin M (IgM) class and various IgG subclasses. To clarify this issue, a murine hybridoma secreting IgM monoclonal antibody (MAb) specific for the O polysaccharide of Pseudomonas aeruginosa serogroup O6 LPS was class switched, by sib selection, to produce an IgG3 MAb with identical specificity and variable region heavy and light chain nucleotide sequences. This IgG3-secreting cell line was further switched to the production of O-specific, variable region-identical IgG1, IgG2b, and IgG2a MAbs. Functional comparisons of these LPS-specific IgM and IgG MAb isotypes revealed similar LPS binding, opsonic, and protective activities. Relatively minor isotype-related differences in levels of efficiency of MAb-mediated, complement-dependent opsonophagocytic killing (IgM > IgG2a > IgG3 > IgG2b > > IgG1) were not associated with corresponding differences in in vivo functions. These findings, in conjunction with previously published data, support a cautious approach to generic conclusions regarding the immunotherapeutic superiority of LPS-specific antibodies belonging to either the IgM or IgG class or to a particular IgG subclass. PMID:7591089

  13. Enzyme immuno assay for the detection of virus specific IgG and IgM antibody in patients with haemorrhagic fever with renal syndrome.

    PubMed

    Ivanov, A P; Tkachenko, E A; Petrov, V A; Pashkov, A J; Dzagurova, T K; Vladimirova, T P; Voronkova, G M; van der Groen, G

    1988-01-01

    Consecutive serum samples collected from 235 patients with Haemorrhagic Fever with Renal Syndrome (HFRS), between two days and two years after onset of disease, have been analysed for the presence of IgG and IgM type of antibodies specific for Hanta-viruses. The sera were screened in parallel by a newly developed indirect Immuno Enzyme Assay (EIA) in parallel with Indirect Immunofluorescent Antibody Assay (IFA). In both tests the Hantaan virus strain 76-118 was used as the antigen. The EIA was much more sensitive than the IFA test for the detection of IgM type antibodies. With the indirect EIA IgM type antibodies against Hantaan virus 76-118 have been detected in HFRS patient's sera from the second day of illness indicating the usefulness of this test for the early serological diagnosis of this disease.

  14. Reduced IgG anti-small nuclear ribonucleoprotein autoantibody production in systemic lupus erythematosus patients with positive IgM anti-cytomegalovirus antibodies

    PubMed Central

    Palafox Sánchez, Claudia Azucena; Satoh, Minoru; Chan, Edward KL; Carcamo, Wendy C; Muñoz Valle, José Francisco; Orozco Barocio, Gerardo; Oregon Romero, Edith; Navarro Hernández, Rosa Elena; Salazar Páramo, Mario; Cabral Castañeda, Antonio; Vázquez del Mercado, Mónica

    2009-01-01

    Introduction Systemic lupus erythematosus is characterized by production of autoantibodies to RNA or DNA–protein complexes such as small nuclear ribonucleoproteins (snRNPs). A role of Epstein–Barr virus in the pathogenesis has been suggested. Similar to Epstein–Barr virus, cytomegalovirus (CMV) infects the majority of individuals at a young age and establishes latency with a potential for reactivation. Homology of CMV glycoprotein B (UL55) with the U1snRNP-70 kDa protein (U1–70 k) has been described; however, the role of CMV infection in production of anti-snRNPs is controversial. We investigated the association of CMV serology and autoantibodies in systemic lupus erythematosus. Methods Sixty-one Mexican patients with systemic lupus erythematosus were tested for CMV and Epstein–Barr virus serology (viral capsid antigen, IgG, IgM) and autoantibodies by immunoprecipitation and ELISA (IgG and IgM class, U1RNP/Sm, U1–70 k, P peptide, rheumatoid factor, dsDNA, β2-glycoprotein I). Results IgG anti-CMV and IgM anti-CMV were positive in 95% (58/61) and 33% (20/61), respectively, and two cases were negative for both. Clinical manifestation and autoantibodies in the IgM anti-CMV(+) group (n = 20) versus the IgM anti-CMV(-)IgG (+) (n = 39) group were compared. Most (19/20) of the IgM anti-CMV(+) cases were IgG anti-CMV(+), consistent with reactivation or reinfection. IgM anti-CMV was unrelated to rheumatoid factor or IgM class autoantibodies and none was positive for IgM anti-Epstein–Barr virus–viral capsid antigen, indicating that this is not simply due to false positive results caused by rheumatoid factor or nonspecific binding by certain IgM. The IgM anti-CMV(+) group has significantly lower levels of IgG anti-U1RNP/Sm and IgG anti-U1–70 k (P = 0.0004 and P = 0.0046, respectively). This finding was also confirmed by immunoprecipitation. Among the IgM anti-CMV(-) subset, anti-Su was associated with anti-U1RNP and anti-Ro (P < 0.05). High levels of Ig

  15. Multiple Plasmodium falciparum Erythrocyte Membrane Protein 1 Variants per Genome Can Bind IgM via Its Fc Fragment Fcμ.

    PubMed

    Jeppesen, Anine; Ditlev, Sisse Bolm; Soroka, Vladyslav; Stevenson, Liz; Turner, Louise; Dzikowski, Ron; Hviid, Lars; Barfod, Lea

    2015-10-01

    The Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1) adhesive proteins expressed on the surfaces of infected erythrocytes (IEs) are of key importance in the pathogenesis of P. falciparum malaria. Several structurally and functionally defined PfEMP1 types have been associated with severe clinical manifestations, such as cerebral malaria in children and placental malaria in pregnant women. PfEMP1 that can bind the Fc part of IgM (Fcμ) characterizes one such type, although the functional significance of this IgM binding to PfEMP1 remains unclear. In this study, we report the identification and functional analysis of five IgM-binding PfEMP1 proteins encoded by P. falciparum NF54. In addition to the VAR2CSA-type PFL0030c protein, already known to bind Fcμ and to mediate chondroitin sulfate A (CSA)-specific adhesion of IEs in the placenta, we found four PfEMP1 proteins not previously known to bind IgM this way. Although they all contained Duffy binding-like ε (DBLε) domains similar to those in VAR2CSA-type PfEMP1, they did not mediate IE adhesion to CSA, and IgM binding did not shield IEs from phagocytosis of IgG-opsonized IEs. In this way, these new IgM-binding PfEMP1 proteins resemble the rosette-mediating and IgM-binding PfEMP1 HB3VAR06, but none of them mediated formation of rosettes. We could map the capacity for Fc-specific IgM binding to DBLε domains near the C terminus for three of the four PfEMP1 proteins tested. Our study provides new evidence regarding Fc-dependent binding of IgM to PfEMP1, which appears to be a common and multifunctional phenotype.

  16. The pyloric caeca area is a major site for IgM(+) and IgT(+) B cell recruitment in response to oral vaccination in rainbow trout.

    PubMed

    Ballesteros, Natalia A; Castro, Rosario; Abos, Beatriz; Rodríguez Saint-Jean, Sylvia S; Pérez-Prieto, Sara I; Tafalla, Carolina

    2013-01-01

    Although previous studies have characterized some aspects of the immune response of the teleost gut in response to diverse pathogens or stimuli, most studies have focused on the posterior segments exclusively. However, there are still many details of how teleost intestinal immunity is regulated that remain unsolved, including the location of IgM(+) and IgT(+) B cells along the digestive tract and their role during the course of a local stimulus. Thus, in the current work, we have studied the B cell response in five different segments of the rainbow trout (Oncorhynchus mykiss) digestive tract in both naïve fish and fish orally vaccinated with an alginate-encapsulated DNA vaccine against infectious pancreatic necrosis virus (IPNV). IgM(+) and IgT(+) cells were identified all along the tract with the exception of the stomach in naïve fish. While IgM(+) cells were mostly located in the lamina propria (LP), IgT(+) cells were primarily localized as intraepithelial lymphocytes (IELs). Scattered IgM(+) IELs were only detected in the pyloric caeca. In response to oral vaccination, the pyloric caeca region was the area of the digestive tract in which a major recruitment of B cells was demonstrated through both real time PCR and immunohistochemistry, observing a significant increase in the number of both IgM(+) and IgT(+) IELs. Our findings demonstrate that both IgM(+) and IgT(+) respond to oral stimulation and challenge the paradigm that teleost IELs are exclusively T cells. Unexpectedly, we have also detected B cells in the fat tissue associated to the digestive tract that respond to vaccination, suggesting that these cells surrounded by adipocytes also play a role in mucosal defense.

  17. Targeting Pathogenic Post-Ischemic Self-Recognition by Natural IgM to Protect Against Post-Transplant Cardiac Reperfusion Injury

    PubMed Central

    Atkinson, Carl; Qiao, Fei; Yang, Xiaofeng; Zhu, Peng; Reaves, Nicholas; Kulik, Liudmila; Goddard, Martin; Holers, V. Michael; Tomlinson, Stephen

    2015-01-01

    Background Natural IgM antibodies represent a class of innate pattern recognition receptors that recognize danger associated molecular patterns expressed on stressed or dying cells. They play important roles in tissue homeostasis by disposing of pre-necrotic cells and suppressing inflammation. However, ischemic insult leads to a pathogenic level of IgM binding and complement activation, resulting in inflammation and injury. We investigate the role of self-reactive IgM in the unique setting of transplantation, where the donor organ undergoes both cold and warm ischemia, and global ischemic insult. Methods and Results By transplanting hearts from wild-type donor mice into antibody-deficient mice reconstituted with specific self-reactive IgM mAbs, we identified neoepitopes expressed post-transplant, and demonstrated a key role for IgM recognition of these epitopes in graft injury. With this information, we developed and characterized a therapeutic strategy that exploited the post-ischemia recognition system of natural antibodies. Based on neoepitope identification, we constructed an anti-annexin-IV single chain antibody (scFv) and an scFv linked to Crry, an inhibitor of C3 activation (scFv-Crry). In an allograft transplant model, in which recipients contain a full natural antibody repertoire, both constructs blocked graft IgM binding and complement activation, and significantly reduced graft inflammation and injury. Furthermore, scFv-Crry specifically targeted to the transplanted heart and, unlike complement deficiency, did not affect immunity to infection, an important consideration for immunosuppressed transplant recipients. Conclusions We identified pathophysiologically important epitopes expressed within the heart post-transplant, and describe a novel translatable strategy for targeted complement inhibition that has several advantages over currently available approaches. PMID:25825397

  18. Evaluation of commercially available diagnostic tests for the detection of dengue virus NS1 antigen and anti-dengue virus IgM antibody.

    PubMed

    Hunsperger, Elizabeth A; Yoksan, Sutee; Buchy, Philippe; Nguyen, Vinh Chau; Sekaran, Shamala Devi; Enria, Delia A; Vazquez, Susana; Cartozian, Elizabeth; Pelegrino, Jose L; Artsob, Harvey; Guzman, Maria G; Olliaro, Piero; Zwang, Julien; Guillerm, Martine; Kliks, Susie; Halstead, Scott; Peeling, Rosanna W; Margolis, Harold S

    2014-10-01

    Commercially available diagnostic test kits for detection of dengue virus (DENV) non-structural protein 1 (NS1) and anti-DENV IgM were evaluated for their sensitivity and specificity and other performance characteristics by a diagnostic laboratory network developed by World Health Organization (WHO), the UNICEF/UNDP/World Bank/WHO Special Programme for Research and Training in Tropical Diseases (TDR) and the Pediatric Dengue Vaccine Initiative (PDVI). Each network laboratory contributed characterized serum specimens for the panels used in the evaluation. Microplate enzyme-linked immunosorbent assay (ELISA) and rapid diagnostic test (RDT formats) were represented by the kits. Each ELISA was evaluated by 2 laboratories and RDTs were evaluated by at least 3 laboratories. The reference tests for IgM anti-DENV were laboratory developed assays produced by the Armed Forces Research Institute for Medical Science (AFRIMS) and the Centers for Disease Control and Prevention (CDC), and the NS1 reference test was reverse transcriptase polymerase chain reaction (RT-PCR). Results were analyzed to determine sensitivity, specificity, inter-laboratory and inter-reader agreement, lot-to-lot variation and ease-of-use. NS1 ELISA sensitivity was 60-75% and specificity 71-80%; NS1 RDT sensitivity was 38-71% and specificity 76-80%; the IgM anti-DENV RDTs sensitivity was 30-96%, with a specificity of 86-92%, and IgM anti-DENV ELISA sensitivity was 96-98% and specificity 78-91%. NS1 tests were generally more sensitive in specimens from the acute phase of dengue and in primary DENV infection, whereas IgM anti-DENV tests were less sensitive in secondary DENV infections. The reproducibility of the NS1 RDTs ranged from 92-99% and the IgM anti-DENV RDTs from 88-94%. PMID:25330157

  19. Anti-α-glucose-based glycan IgM antibodies predict relapse activity in multiple sclerosis after the first neurological event

    PubMed Central

    Freedman, MS; Laks, J; Dotan, N; Altstock, RT; Dukler, A; Sindic, CJM

    2009-01-01

    Background There is no specific serum-based biomarker for the diagnosis or prognosis of relapsing-remitting multiple sclerosis (RRMS). Objective We investigated whether levels of IgM antibodies to Glc(α1,4)Glc(α) (GAGA4) or to a panel of four glucose-based glycans could differentiate MS from other neurological diseases (OND) or predict risk of early relapse following first presentation (FP) of RRMS. Methods Retrospective analysis of 440 sera samples of three cohorts: A) FP-RRMS (n = 44), OND (n = 44); B) FP-RRMS (n = 167), OND (n = 85); and C) FP (n = 100). Anti-GAGA4 IgM levels were measured by enzyme immunoassay in cohort-A and cohort-B. Cohort-C IgM antibodies to glucosebased glycan panel were measured by immunofluorescence. Results FP-RRMS had higher levels of anti-GAGA4 IgM than OND patients (cohort-A, P = 0.01; cohort-B, P = 0.0001). Sensitivity and specificity were 27% and 97% for cohort-A; and 26% and 90% for cohort-B, respectively. In cohort-C, 58 patients experienced early relapse (<24 months), 31 had late relapse (≥24 months), and 11 did not experience second attack during follow-up. Kaplan– Meier curves demonstrated decrease in time to next relapse for patients positive for the antibody panel (P = 0.02, log rank). Conclusions Serum anti-GAGA4 IgM discerns FP-RRMS patients from OND patients. Higher levels of serum anti-α-glucose IgM in FP patients predict imminent early relapse. PMID:19324980

  20. Analysis of subsets of B cells, Breg, CD4Treg and CD8Treg cells in adult patients with primary selective IgM deficiency

    PubMed Central

    Louis, Ankmalika Gupta; Agrawal, Sudhanshu; Gupta, Sudhir

    2016-01-01

    Primary selective IgM deficiency (SIGMD) is a rare and recently IUIS-recognized primary immunodeficiency disease with increased susceptibility to infections, allergy, and autoimmune diseases. The pathogenesis of selective IgM remains unclear. The objective of the study was to understand the pathogenesis of selective IgM deficiency via a comprehensive analysis of subsets of B cells, naïve and memory subsets of CD4+ and CD8+ T cells, and Breg, CD4Treg, and CD8Treg cells. Twenty adult patients with SIGMD (serum IgM 4 mg/dl-32 mg/dl) and age-and gender-matched healthy controls were studied. Naïve B cells, transitional B cells, marginal zone B cells, germinal center B cells, IgM memory B cells, switched memory B cells, plasmablasts, CD21low B cells, B1 cells, CXCR3+ naive and memory B cells; naïve, central memory, and effector memory subsets of CD4+ and CD8+ T cells, and CD4Treg, CD8Treg and Breg were phenotypically analyzed using multicolor flow cytometry. A significant increase in CD21low, IgM memory B cells, Breg and CD8Treg, and a significant decreased in germinal center B cells, and CXCR3+ naïve and memory B cells were observed in SIGMD. These alterations in subsets of B cells, and Breg and CD8Treg cells may play a role in the pathogenesis of SIGMD. PMID:27168952

  1. [[Virus-like particle-based immunoglobulin M capture enzyme-linked immunosorbent assay for the detection of IgM antibodies against Chikungunya virus].

    PubMed

    Li, Jian-dong; Zhang, Quan-fu; Zhang, Shuo; Li, Chuan; Liu, Qin-zhi; Liang, Mi-fang; Li, De-xin

    2014-11-01

    To establish a MacELISA method for the detection of IgM antibodies against Chikungunya virus (CHIKV), we prepared virus like particle (VLP) antigens of CHIKV using the whole structural protein C-E3-E2-6K-E1 encoding gene with a baculovirus expression system in Sf9 insect cells. The VLPs were purified and used to immunize Kunming mice. Then, polyclonal antibodies were purified from the samples of ascites with a protein G HiTrap SP column and labeled with horseradish peroxidase. A MacELISA method for the detection of IgM antibodies against CHIKV was assembled with goat anti-human IgM antibody, VLP antigens and an enzyme-labeled polyclonal antibody. The results were evaluated with a serum panel containing serum samples from laboratory-confirmed CHIK, HFRS patients, healthy donors, and commercially available CHIKV IgM as a quality control. It was shown that the MacELISA had a specificity of 99% (99/100), the coefficients of variation (CoV) within a plate were <10%, and the CoV of different ELISA plates in terms of the plate variation coefficient was <15%. A comparative analysis was performed to compare the current method against a commercial CHIKV IgM antibody detection kit for IIFA-IgM. The detection limit of MacELISA was significantly lower than that of the IIFA-IgM commercial kit (P< 0.0001). Here, we demonstrate that the VLP-based MacELISA is a promising tool for the early diagnosis and epidemiological investigation of CHIKV infection, with a high level of sensitivity and specificity for the detection of IgM antibodies against CHIKV.

  2. Clinicopathologic Characterization of Diffuse-Large-B-Cell Lymphoma with an Associated Serum Monoclonal IgM Component

    PubMed Central

    Scarpino, Stefania; Salerno, Gerardo; Tatarelli, Caterina; Talerico, Caterina; Lombardi, Mariangela; Monarca, Bruno; Amadori, Sergio; Ruco, Luigi

    2014-01-01

    Recently, diffuse-large-B-cell lymphoma (DLBCL) associated with serum IgM monoclonal component (MC) has been shown to be a very poor prognostic subset although, detailed pathological and molecular data are still lacking. In the present study, the clinicopathological features and survival of IgM-secreting DLBCL were analyzed and compared to non-secreting cases in a series of 151 conventional DLBCL treated with R-CHOP. IgM MC was detected in 19 (12.5%) out of 151 patients at disease onset. In 17 of these cases secretion was likely due to the neoplastic clone, as suggested by the expression of heavy chain IgM protein in the cytoplasm of tumor cells. In IgM-secreting cases immunoblastic features (p<.0001), non-GCB-type (p = .002) stage III-IV(p = .003), ≥2 extra nodal sites (p<.0001), bone-marrow (p = .002), central-nervous-system (CNS) involvement at disease onset or relapse (p<.0001), IPI-score 3–5 (p = .009) and failure to achieve complete remission (p = .005), were significantly more frequent. FISH analyses for BCL2, BCL6 and MYC gene rearrangements detected only two cases harboring BCL2 gene translocation and in one case a concomitant BCL6 gene translocation was also observed. None of the IgM-secreting DLBCL was found to have L265P mutation of MYD88 gene. Thirty-six month event-free (11.8% vs 66.4% p<.0001), progression-free (23.5% vs 75.7%, p<.0001) and overall (47.1% vs 74.8%, p<.0001) survivals were significantly worse in the IgM-secreting group. In multivariate analysis IgM-secreting (p = .005, expB = 0.339, CI = 0.160-0.716) and IPI-score 3–5 (p = .010, expB = 0.274, CI = 0.102–0.737) were the only significant factors for progression-free-survival. Notably, four relapsed patients, who were treated with salvage immmunochemotherapy combined with bortezomib or lenalidomide, achieved lasting remission. Our data suggests that IgM-secreting cases are a distinct subset of DLBCL, originating from activated

  3. Clonidine inhibits anti-non-Gal IgM xenoantibody elicited in multiple pig-to-primate models

    PubMed Central

    Stewart, John M.; Tarantal, Alice F.; Hawthorne, Wayne J.; Salvaris, Evelyn J.; O’Connell, Philip J.; Nottle, Mark B.; d’Apice, Anthony J. F.; Cowan, Peter J.; Kearns-Jonker, Mary

    2016-01-01

    Background Survival of vascularized xenografts is dependent on pre-emptive inhibition of the xenoantibody response against galactosyltransferase knockout (GTKO) porcine organs. Our analysis in multiple GTKO pig-to-primate models of xenotransplantation has demonstrated that the anti-non-gal-α-1,3-gal (anti-non-Gal) xenoanti-body response displays limited structural diversity. This allowed our group to identify an experimental compound which selectively inhibited induced anti-non-Gal IgM xenoantibodies. However, because this compound had an unknown safety profile, we extended this line of research to include screening small molecules with known safety profiles allowing rapid advancement to large animal models. Methods The NIH clinical collections of small molecules were screened by ELISA for their ability to inhibit xenoantibody binding to GTKO pig endothelial cells. Serum collected from non-immunosuppressed rhesus monkeys at day 14 post-injection with GTKO pig endothelial cells was utilized as a source of elicited xenoantibody for initial screening. Virtual small molecule screening based on xenoanti-body structure was used to assess the likelihood that the identified small molecules bound xenoantibody directly. As a proxy for selectivity, ELISAs against tetanus toxoid and the natural antigens laminin, thyroglobulin, and single-stranded DNA (ssDNA) were utilized to assess the ability of the identified reagents to inhibit additional antibody responses. The identified inhibitory small molecules were further tested for their ability to inhibit xenoantibody elicited in multiple settings, including rhesus monkeys pre-treated with an anti-non-Gal selective anti-idiotypic antibody, non-immunosuppressed rhesus monkeys immunized with wild-type fetal pig isletlike cell clusters, and non-immunosuppressed baboons transplanted with GTKO multiple transgenic pig kidneys. Results Four clinically relevant small molecules inhibited anti-non-Gal IgM binding to GTKO pig endothelial

  4. Profound specific suppression by antigen of persistent IgM, IgG, and IgE antibody production.

    PubMed Central

    Dintzis, H M; Dintzis, R Z

    1992-01-01

    Ongoing, high-titer T-cell-dependent immune responses in adult mice, consisting of IgM, IgG, and IgE anti-fluorescein antibodies, can be specifically and substantially reduced (90-99%) when the mice are injected with appropriate doses of fluoresceinated dextran of defined molecular weight and hapten valence. This suppressive form of the antigen is nontoxic and specific, as responses to other antigens are unaffected. The suppression is long lasting and reduces high-affinity antibodies most markedly. Moreover, plasma cell secretion of specific antibody is virtually eliminated. This demonstrates that the reduction in antibody titer is not simply due to masking of serum antibody by the suppressive polymer. The results are discussed with reference to proposed models of B-cell and T-cell tolerance. Extension of these findings to disease-related immunogens may yield effective antigen-specific treatments of human allergy and autoimmune diseases. PMID:1736295

  5. Immune dysfunction in Rett syndrome patients revealed by high levels of serum anti-N(Glc) IgM antibody fraction.

    PubMed

    Papini, Anna Maria; Nuti, Francesca; Real-Fernandez, Feliciana; Rossi, Giada; Tiberi, Caterina; Sabatino, Giuseppina; Pandey, Shashank; Leoncini, Silvia; Signorini, Cinzia; Pecorelli, Alessandra; Guerranti, Roberto; Lavielle, Solange; Ciccoli, Lucia; Rovero, Paolo; De Felice, Claudio; Hayek, Joussef

    2014-01-01

    Rett syndrome (RTT), a neurodevelopmental disorder affecting exclusively (99%) female infants, is associated with loss-of-function mutations in the gene encoding methyl-CpG binding protein 2 (MECP2) and, more rarely, cyclin-dependent kinase-like 5 (CDKL5) and forkhead box protein G1 (FOXG1). In this study, we aimed to evaluate the function of the immune system by measuring serum immunoglobulins (IgG and IgM) in RTT patients (n = 53) and, by comparison, in age-matched children affected by non-RTT pervasive developmental disorders (non-RTT PDD) (n = 82) and healthy age-matched controls (n = 29). To determine immunoglobulins we used both a conventional agglutination assay and a novel ELISA based on antibody recognition by a surrogate antigen probe, CSF114(Glc), a synthetic N-glucosylated peptide. Both assays provided evidence for an increase in IgM titer, but not in IgG, in RTT patients relative to both healthy controls and non-RTT PDD patients. The significant difference in IgM titers between RTT patients and healthy subjects in the CSF114(Glc) assay (P = 0.001) suggests that this procedure specifically detects a fraction of IgM antibodies likely to be relevant for the RTT disease. These findings offer a new insight into the mechanism underlying the Rett disease as they unveil the possible involvement of the immune system in this pathology. PMID:25389532

  6. Detection of IgM antibodies against a recombinant nucleocapsid protein of canine distemper virus in dog sera using a dot-blot assay.

    PubMed

    Barben, G; Stettler, M; Jaggy, A; Vandevelde, M; Zurbriggen, A

    1999-03-01

    A dot-blot assay for the detection of IgM antibodies (ABs) against canine distemper virus (CDV) in canine serum is described. The diagnostic potential of this technique was evaluated by analysing sera from three test groups: (i) specific pathogen-free (SPF) beagle dogs experimentally infected with virulent CDV; (ii) SPF dogs immunized with a combined vaccine containing CDV, and (iii) SPF dogs immunized with a CDV-free vaccine. As antigen for the dot-blot assay we used the recombinant nucleocapsid protein (N protein) of the virulent A75/17 CDV strain. All 12 dogs of group 1, infected with virulent CDV, showed detectable CDV-specific IgM levels in their serum. All dogs of group 2 were also positive for anti-CDV IgM after the first immunization with the CDV-containing vaccine. The four dogs immunized with a CDV-free vaccine (group iii) remained negative throughout the course of the experiment. From these results, we conclude that the IgM detection test, which requires only a single serum sample, is a useful method for diagnosing current or recent CDV infection in CDV-infected or CDV-immunized dogs under experimental conditions. PMID:10216448

  7. Development of a particle agglutination assay system for detecting Japanese encephalitis virus-specific human IgM, using hydroxyapatite-coated nylon beads.

    PubMed

    Yamamoto, Akira; Nakayama, Mikio; Kurosawa, Yae; Sugo, Ken; Karasawa, Hideharu; Ogawa, Tetsuro; Takasaki, Tomohiko; Tashiro, Masato; Kurane, Ichiro

    2002-07-01

    Japanese encephalitis virus-specific IgM is a reliable indicator for serodiagnosis of Japanese encephalitis. A particle agglutination (PA) assay system was developed to detect anti-Japanese encephalitis virus IgM in human serum samples. The newly developed PA assay consisted of hydroxyapatite-coated nylon beads and V-bottom 96-well microplates. Hydroxyapatite-coated nylon beads were coated with Japanese encephalitis virus antigens. Japanese encephalitis virus antigen-coated, hydroxyapatite-coated nylon beads agglutinated in the IgM-captured wells when anti-Japanese encephalitis virus IgM-positive serum samples were used. A button pattern was formed at the bottom of the wells when anti-Japanese encephalitis virus IgM-negative serum samples were used. Thirty anti-Japanese encephalitis virus IgM-positive serum samples from Japanese encephalitis-confirmed cases were tested by the PA assay. All these serum samples were determined to be Japanese encephalitis virus IgM-positive. IgM titers determined by the PA assay corresponded to those determined by enzyme-linked immunosorbent assay. The titers were consistent in two independent PA assays. These results indicate that the newly developed PA assay is a reliable method for detecting anti-Japanese encephalitis virus IgM in human serum samples and that this assay will be a suitable diagnostic system especially in rural areas of Asia.

  8. Assessment of the diagnostic performance of the IDS-iSYS tests for toxo IgG, toxo IgM and avidity.

    PubMed

    Levigne, Pauline; Peyron, François; Wallon, Martine

    2016-10-01

    When acquired during pregnancy toxoplasmosis can have devastating consequences on the fetus. As maternal infection is in the majority of cases subclinical, the diagnosis of toxoplasmosis relies on serological tests for the detection of IgG, IGM and the mesure of IgG avidity. We evaluated the performance of IDS-iSYS a new automatized instrument based on chemiluminescence for the diagnosis of the disease. Our study was based on non-selected samples received in our laboratory either for the determination of serological status or for distinguishing acute from chronic infection. Seven hundred eighty three samples were enrolled in the study. Compared with Architect IgG and IgM assays, the sensitivity and specificity were respectively 99% and 99% for IgG, and 75% and 97% for IgM. We observed higher remaining titers for IDS iSYS IgG, which could reduce the proportions of patients who have to be retested because of doubtful titers. IgM detection and avidity scored equivalent performance with both methods. This automate appears to be a reliable and easy-to-use tool for diagnosing toxoplasmosis in different clinical settings.

  9. IgM, IgA and IgG producing cells in cerebrospinal fluid and peripheral blood in multiple sclerosis.

    PubMed Central

    Henriksson, A; Kam-Hansen, S; Link, H

    1985-01-01

    The protein A plaque assay was used to enumerate IgM, IgA and IgG producing cells per 20 X 10(3) lymphocytes in cerebrospinal fluid (CSF) and peripheral blood (PB) from 37 patients with multiple sclerosis (MS) and in PB from healthy controls. Fifty-seven percent of the MS patients displayed in CSF cells producing IgM, 70% IgA and 89% IgG. IgM or IgA producing cells predominated in CSF from 10 patients, IgG in 27. Immunoglobulin producing cells were often present when the corresponding CSF Ig index was normal, confirming that enumeration of Ig producing cells is a more sensitive variable of the intrathecal immune status. No Ig producing cells were found in CSF from four patients with tension headache, indicating absence of intrathecal Ig synthesis in healthy individuals. The patients with MS had higher numbers of IgM, IgA and IgG producing cells in PB than healthy controls, confirming occurrence of an extrathecal B cell response in MS. Active and stable MS patients did not differ regarding Ig producing cells in CSF nor in PB, which speaks in favour of continuous immune activity within as well as outside the CNS independent of clinical symptoms. PMID:4064372

  10. Search for QSOS Suitable for Subsequent Observation of he II 304 Absorption Arising in the IGM, Ly-Alpha and ... PART2

    NASA Astrophysics Data System (ADS)

    Tytler, David

    1991-07-01

    THIS IS PART TWO OF PROPOSAL 3801, CONTAINING 55 TARGETS OF PRIORITY 7,8 AND 9. EXCEPT FOR TARGETS, THIS PROPOSAL IS IDENTICAL TO 3801, WHICH HAS 87 TARGETS OF PRIORITY 3,4,5 AND 6. Ultraviolet images will be obtained in snapshot mode of the 500 known high-redshift (z > 2.8) QSOs in order to identify the few (about 20) targets which have sufficient ultraviolet flux for subsequent FOC/FOS or GHRS observations of He II 304. The detection of absorption by the Helium II Lyman-alpha line at 304 A, one of the most exciting prospects of the HST, will provide the first direct detection of the diffuse intergalactic medium (IGM). The absence of Gunn-Peterson H I 1215 absorption shows that the IGM is hot and/or of very low density, thus He I 584 is not expected to be observable. He II 304--the most promising line--should be observable from three sources: the diffuse IGM, the discrete Ly-alpha clouds, and the much rarer metal line absorption systems. The Gunn-Peterson continuum optical depth is not well constrained by models (range 0.3-3000). The mere detection of only one QSO below 304 A would rule out many models, limiting the IGM density, temperature, and ionization mechanisms. Similarly the total absence of flux from several targets would rule out other models.

  11. Search for QSOS Suitable for Subsequent Observation of he II 304 Absorption Arising in the IGM, Ly-Alpha and ... PART3

    NASA Astrophysics Data System (ADS)

    Tytler, David

    1991-07-01

    THIS IS PART Three OF PROPOSAL 3801. IT CONTAINS 40 TARGETS OF HIGH PRIORITY. EXCEPT FOR TARGETS, THIS PROPOSAL IS IDENTICAL TO 3801. Targets are in order of decreasing priority. Priority matters a lot. Best targets are listed first. Ultraviolet images will be obtained in snapshot mode of the 500 known high-redshift (z > 2.8) QSOs in order to identify the few (about 20) targets which have sufficient ultraviolet flux for subsequent FOC/FOS or GHRS observations of He II 304. The detection of absorption by the Helium II Lyman-alpha line at 304 A, one of the most exciting prospects of the HST, will provide the first direct detection of the diffuse intergalactic medium (IGM). The absence of Gunn-Peterson H I 1215 absorption shows that the IGM is hot and/or of very low density, thus He I 584 is not expected to be observable. He II 304--the most promising line--should be observable from three sources: the diffuse IGM, the discrete Ly-alpha clouds, and the much rarer metal line absorption systems. The Gunn-Peterson continuum optical depth is not well constrained by models (range 0.3-3000). The mere detection of only one QSO below 304 A would rule out many models, limiting the IGM density, temperature, and ionization mechanisms. Similarly the total absence of flux from several targets would rule out other models.

  12. The effect of various metal ions and chelating agents on the formation of noncovalently and covalently linked IgM polymers.

    PubMed

    Eskeland, T

    1977-01-01

    The influence of various concentrations of Mg2+, Ca2+, Mn2+, Fe2+, Fe3+, Co2+, Ni2+, Cu2+, Zn2+, Cd2+, Hg2+, EDTA, or 8-hydroxyquinoline-5-sulfonic acid on the formation in vitro of polymeric IgM from reduced and dissociated IgM has been investigated. 4-200muM Zn, Cd, or Hg ions in the dialysis buffer used for reassociation resulted in the formation of a homogeneous polymer peak sedimenting as 19S IgM, whereas the other metal ions resulted in the formation of polymers sedimenting mostly as tetramers, pentamers (19S IgM), and hexamers. Dialysis in the presence of 1 and 3mM EDTA or 8-hydroxyquinoline-5-sulfonic acid gave slight polymer formation; 10 mM of the agents gave none. Ca, Mg, Fe, Ni, Zn, Cd, or Hg ions regularly gave only noncovalently linked polymers, whereas Cu ions in particular, but also Co and, to a minor extent, Mn ions catalyzed the formation of covalently linked polymers. Experiments performed with Mg, Ca, Ni, and Zn ions or with Cu and Zn ions in the same buffer during reassociation showed that Zn ions inhibit the effect of the other ions on polymer assembly and covalent stabilization.

  13. Long-lived antigen-induced IgM plasma cells demonstrate somatic mutations and contribute to long-term protection

    PubMed Central

    Bohannon, Caitlin; Powers, Ryan; Satyabhama, Lakshmipriyadarshini; Cui, Ang; Tipton, Christopher; Michaeli, Miri; Skountzou, Ioanna; Mittler, Robert S.; Kleinstein, Steven H.; Mehr, Ramit; Lee, Frances Eun-Yun; Sanz, Ignacio; Jacob, Joshy

    2016-01-01

    Long-lived plasma cells are critical to humoral immunity as a lifelong source of protective antibodies. Antigen-activated B cells—with T-cell help—undergo affinity maturation within germinal centres and persist as long-lived IgG plasma cells in the bone marrow. Here we show that antigen-specific, induced IgM plasma cells also persist for a lifetime. Unlike long-lived IgG plasma cells, which develop in germinal centres and then home to the bone marrow, IgM plasma cells are primarily retained within the spleen and can develop even in the absence of germinal centres. Interestingly, their expressed IgV loci exhibit somatic mutations introduced by the activation-induced cytidine deaminase (AID). However, these IgM plasma cells are probably not antigen-selected, as replacement mutations are spread through the variable segment and not enriched within the CDRs. Finally, antibodies from long-lived IgM plasma cells provide protective host immunity against a lethal virus challenge. PMID:27270306

  14. Distinct Differentiation Programs Triggered by IL-6 and LPS in Teleost IgM(+) B Cells in The Absence of Germinal Centers.

    PubMed

    Abós, Beatriz; Wang, Tiehui; Castro, Rosario; Granja, Aitor G; Leal, Esther; Havixbeck, Jeffrey; Luque, Alfonso; Barreda, Daniel R; Secombes, Chris J; Tafalla, Carolina

    2016-01-01

    Although originally identified as a B cell differentiation factor, it is now known that mammalian interleukin-6 (IL-6) only regulates B cells committed to plasma cells in response to T-dependent (TD) antigens within germinal centers (GCs). Even though adaptive immunity is present in teleost fish, these species lack lymph nodes and GCs. Thus, the aim of the present study was to establish the role of trout IL-6 on B cells, comparing its effects to those induced by bacterial lipopolysaccharide (LPS). We demonstrate that the effects of teleost IL-6 on naïve spleen B cells include proliferation, activation of NF-κB, increased IgM secretion, up-regulation of Blimp1 transcription and decreased MHC-II surface expression that point to trout IL-6 as a differentiation factor for IgM antibody-secreting cells (ASCs). However, LPS induced the secretion of IgM without up-regulating Blimp1, driving the cells towards an intermediate activation state in which antigen presenting mechanisms are elicited together with antibody secretion and expression of pro-inflammatory genes. Our results reveal that, in trout, IL-6 is a differentiation factor for B cells, stimulating IgM responses in the absence of follicular structures, and suggest that it was after follicular structures appeared that this cytokine evolved to modulate TD responses within the GC. PMID:27481356

  15. Characterization of IgA and IgM binding and internalization by surface-expressed human Fcα/μ receptor.

    PubMed

    Yoo, Esther M; Trinh, K Ryan; Lim, Hana; Wims, Letitia A; Morrison, Sherie L

    2011-09-01

    The Fcα/μ receptor (Fcα/μR) is an unusual Fc receptor in that it binds to two different antibody isotypes, IgA and IgM. This receptor is of interest because it is thought to be involved in the capture of IgA- and IgM-immune complexes and antigen presentation. To further characterize this receptor, we were able to stably express human Fcα/μR on the surface of the 293T cell line. Using this system, we determined the affinity of the interactions of the receptor with IgA and IgM, which led to novel insights including the important finding that IgM polymers can bind to human Fcα/μR in the absence of J chain. This is in contrast to the polymeric immunoglobulin receptor (pIgR), which requires the presence of J chain to bind to polymeric IgA and IgM. The dissociation constants (K(d)) of all of the different human IgA isotypes and allotypes for human Fcα/μR were determined, and we show that the N-linked glycans on IgA1 are not required for binding to the receptor. In addition, we demonstrate that IgA can be rapidly internalized by human Fcα/μR in the presence of cross-linking antibody.

  16. Hemolytic anemia related to an IgM autoantibody to phosphatidylcholine that binds in vitro to stored and to bromelain-treated human erythrocytes.

    PubMed

    Cabral, A R; Cabiedes, J; Alarcón-Segovia, D

    1990-12-01

    Both normal and autoimmune mice have IgM natural autoantibodies to bromelain-treated erythrocytes in which phosphatidylcholine (PTC) becomes exposed. At one stage this antibody may participate in the genesis of autoimmune hemolytic anemia in the NZB mouse. We have recently studied a patient with hemolytic anemia who had persistently high serum titers of IgM anticardiolipin antibodies (aCL) that were also demonstrated in a hemolysate of his erythrocytes obtained at the time of the anemia. We affinity-purified the antibody and sought its binding to normal human bromelain-treated erythrocytes (BrE) because of the IgM isotype of the antibody, since cardiolipin is not a constituent of the erythrocyte wall, and because the anionic phospholipids, with which aCL are known to cross-react, are not located at the outer leaflet of the erythrocyte membrane. We found binding of the antibody to HBrE in their hemolysates and by flow cytometry. We also demonstrated that the aCL cross-reacted extensively with PTC, as well as with other anionic or zwitterionic phospholipids. The purified IgM antibody lysed BrE in the presence of complement and also bound to in vitro-aged erythrocytes. Because this patient had no other evidence of systemic lupus erythematosus or any other autoimmune condition, his disease may represent a variant of the recently described primary antiphospholipid syndrome.

  17. Agglutinating mouse IgG3 compares favourably with IgMs in typing of the blood group B antigen: Functionality and stability studies

    PubMed Central

    Klaus, Tomasz; Bzowska, Monika; Kulesza, Małgorzata; Kabat, Agnieszka Martyna; Jemioła-Rzemińska, Małgorzata; Czaplicki, Dominik; Makuch, Krzysztof; Jucha, Jarosław; Karabasz, Alicja; Bereta, Joanna

    2016-01-01

    Mouse immunoglobulins M (IgMs) that recognize human blood group antigens induce haemagglutination and are used worldwide for diagnostic blood typing. Contrary to the current belief that IgGs are too small to simultaneously bind antigens on two different erythrocytes, we obtained agglutinating mouse IgG3 that recognized antigen B of the human ABO blood group system. Mouse IgG3 is an intriguing isotype that has the ability to form Fc-dependent oligomers. However, F(ab′)2 fragments of the IgG3 were sufficient to agglutinate type B red blood cells; therefore, IgG3-triggered agglutination did not require oligomerization. Molecular modelling indicated that mouse IgG3 has a larger range of Fab arms than other mouse IgG subclasses and that the unique properties of mouse IgG3 are likely due to the structure of its hinge region. With a focus on applications in diagnostics, we compared the stability of IgG3 and two IgMs in formulated blood typing reagents using an accelerated storage approach and differential scanning calorimetry. IgG3 was much more stable than IgMs. Interestingly, the rapid decrease in IgM activity was caused by aggregation of the molecules and a previously unknown posttranslational proteolytic processing of the μ heavy chain. Our data point to mouse IgG3 as a potent diagnostic tool. PMID:27484487

  18. Agglutinating mouse IgG3 compares favourably with IgMs in typing of the blood group B antigen: Functionality and stability studies.

    PubMed

    Klaus, Tomasz; Bzowska, Monika; Kulesza, Małgorzata; Kabat, Agnieszka Martyna; Jemioła-Rzemińska, Małgorzata; Czaplicki, Dominik; Makuch, Krzysztof; Jucha, Jarosław; Karabasz, Alicja; Bereta, Joanna

    2016-01-01

    Mouse immunoglobulins M (IgMs) that recognize human blood group antigens induce haemagglutination and are used worldwide for diagnostic blood typing. Contrary to the current belief that IgGs are too small to simultaneously bind antigens on two different erythrocytes, we obtained agglutinating mouse IgG3 that recognized antigen B of the human ABO blood group system. Mouse IgG3 is an intriguing isotype that has the ability to form Fc-dependent oligomers. However, F(ab')2 fragments of the IgG3 were sufficient to agglutinate type B red blood cells; therefore, IgG3-triggered agglutination did not require oligomerization. Molecular modelling indicated that mouse IgG3 has a larger range of Fab arms than other mouse IgG subclasses and that the unique properties of mouse IgG3 are likely due to the structure of its hinge region. With a focus on applications in diagnostics, we compared the stability of IgG3 and two IgMs in formulated blood typing reagents using an accelerated storage approach and differential scanning calorimetry. IgG3 was much more stable than IgMs. Interestingly, the rapid decrease in IgM activity was caused by aggregation of the molecules and a previously unknown posttranslational proteolytic processing of the μ heavy chain. Our data point to mouse IgG3 as a potent diagnostic tool. PMID:27484487

  19. Evaluation of Meridian ImmunoCard Mycoplasma test for the detection of Mycoplasma pneumoniae-specific IgM in paediatric patients.

    PubMed

    Matas, L; Domínguez, J; De Ory, F; García, N; Galí, N; Cardona, P J; Hernández, A; Rodrigo, C; Ausina, V

    1998-01-01

    The Meridian ImmunoCard Mycoplasma kit, a 10-min card-based enzyme-linked immunosorbent assay (ELISA) designed to detect immunoglobulin M (IgM) antibodies to Mycoplasma pneumoniae was evaluated. We compared the ImmunoCard with the Fujirebio Serodia Myco II particle agglutination test, as well as with the complement fixation (CF) test to detect M. pneumoniae antibodies in paediatric patients. The ImmunoCard test and Serodia Myco II test agreed in 93.95%, and ImmunoCard test and CF test agreed in 83.51% of the 182 specimens tested. Nine specimens gave negative particle agglutination titres in the acute phase sample, and 28 specimens gave negative CF titres in the acute phase sample, although in the ImmunoCard test they were positive. These results may indicate that the ImmunoCard assay detects lower IgM levels of antibodies than the Serodia Myco II and CF test. The ImmunoCard appears to be a good screening assay test for M. pneumoniae IgM in children in whom M. pneumoniae IgM is found frequently.

  20. Immunohistochemical detection of IgM and IgG in lung tissue of dogs with leptospiral pulmonary haemorrhage syndrome (LPHS)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Leptospiral pulmonary haemorrhage syndrome (LPHS) is a severe form of leptospirosis. Pathogenic mechanisms are poorly understood. Lung tissues from 26 dogs with LPHS, 5 dogs with pulmonary haemorrhage due to other causes and 6 healthy lungs were labelled for IgG, IgM and leptospiral antigens. Three ...

  1. Distinct Differentiation Programs Triggered by IL-6 and LPS in Teleost IgM+ B Cells in The Absence of Germinal Centers

    PubMed Central

    Abós, Beatriz; Wang, Tiehui; Castro, Rosario; Granja, Aitor G.; Leal, Esther; Havixbeck, Jeffrey; Luque, Alfonso; Barreda, Daniel R.; Secombes, Chris J.; Tafalla, Carolina

    2016-01-01

    Although originally identified as a B cell differentiation factor, it is now known that mammalian interleukin-6 (IL-6) only regulates B cells committed to plasma cells in response to T-dependent (TD) antigens within germinal centers (GCs). Even though adaptive immunity is present in teleost fish, these species lack lymph nodes and GCs. Thus, the aim of the present study was to establish the role of trout IL-6 on B cells, comparing its effects to those induced by bacterial lipopolysaccharide (LPS). We demonstrate that the effects of teleost IL-6 on naïve spleen B cells include proliferation, activation of NF-κB, increased IgM secretion, up-regulation of Blimp1 transcription and decreased MHC-II surface expression that point to trout IL-6 as a differentiation factor for IgM antibody-secreting cells (ASCs). However, LPS induced the secretion of IgM without up-regulating Blimp1, driving the cells towards an intermediate activation state in which antigen presenting mechanisms are elicited together with antibody secretion and expression of pro-inflammatory genes. Our results reveal that, in trout, IL-6 is a differentiation factor for B cells, stimulating IgM responses in the absence of follicular structures, and suggest that it was after follicular structures appeared that this cytokine evolved to modulate TD responses within the GC. PMID:27481356

  2. Assessment of the diagnostic performance of the IDS-iSYS tests for toxo IgG, toxo IgM and avidity.

    PubMed

    Levigne, Pauline; Peyron, François; Wallon, Martine

    2016-10-01

    When acquired during pregnancy toxoplasmosis can have devastating consequences on the fetus. As maternal infection is in the majority of cases subclinical, the diagnosis of toxoplasmosis relies on serological tests for the detection of IgG, IGM and the mesure of IgG avidity. We evaluated the performance of IDS-iSYS a new automatized instrument based on chemiluminescence for the diagnosis of the disease. Our study was based on non-selected samples received in our laboratory either for the determination of serological status or for distinguishing acute from chronic infection. Seven hundred eighty three samples were enrolled in the study. Compared with Architect IgG and IgM assays, the sensitivity and specificity were respectively 99% and 99% for IgG, and 75% and 97% for IgM. We observed higher remaining titers for IDS iSYS IgG, which could reduce the proportions of patients who have to be retested because of doubtful titers. IgM detection and avidity scored equivalent performance with both methods. This automate appears to be a reliable and easy-to-use tool for diagnosing toxoplasmosis in different clinical settings. PMID:27450223

  3. Immune Dysfunction in Rett Syndrome Patients Revealed by High Levels of Serum Anti-N(Glc) IgM Antibody Fraction

    PubMed Central

    Papini, Anna Maria; Real-Fernandez, Feliciana; Rossi, Giada; Tiberi, Caterina; Sabatino, Giuseppina; Pandey, Shashank; Lavielle, Solange; Ciccoli, Lucia; Hayek, Joussef

    2014-01-01

    Rett syndrome (RTT), a neurodevelopmental disorder affecting exclusively (99%) female infants, is associated with loss-of-function mutations in the gene encoding methyl-CpG binding protein 2 (MECP2) and, more rarely, cyclin-dependent kinase-like 5 (CDKL5) and forkhead box protein G1 (FOXG1). In this study, we aimed to evaluate the function of the immune system by measuring serum immunoglobulins (IgG and IgM) in RTT patients (n = 53) and, by comparison, in age-matched children affected by non-RTT pervasive developmental disorders (non-RTT PDD) (n = 82) and healthy age-matched controls (n = 29). To determine immunoglobulins we used both a conventional agglutination assay and a novel ELISA based on antibody recognition by a surrogate antigen probe, CSF114(Glc), a synthetic N-glucosylated peptide. Both assays provided evidence for an increase in IgM titer, but not in IgG, in RTT patients relative to both healthy controls and non-RTT PDD patients. The significant difference in IgM titers between RTT patients and healthy subjects in the CSF114(Glc) assay (P = 0.001) suggests that this procedure specifically detects a fraction of IgM antibodies likely to be relevant for the RTT disease. These findings offer a new insight into the mechanism underlying the Rett disease as they unveil the possible involvement of the immune system in this pathology. PMID:25389532

  4. Diagnostic Accuracy of Recombinant Immunoglobulin-like Protein A-Based IgM ELISA for the Early Diagnosis of Leptospirosis in the Philippines

    PubMed Central

    Kitashoji, Emi; Koizumi, Nobuo; Lacuesta, Talitha Lea V.; Usuda, Daisuke; Ribo, Maricel R.; Tria, Edith S.; Go, Winston S.; Kojiro, Maiko; Parry, Christopher M.; Dimaano, Efren M.; Villarama, Jose B.; Ohnishi, Makoto; Suzuki, Motoi; Ariyoshi, Koya

    2015-01-01

    Background Leptospirosis is an important but largely under-recognized public health problem in the tropics. Establishment of highly sensitive and specific laboratory diagnosis is essential to reveal the magnitude of problem and to improve treatment. This study aimed to evaluate the diagnostic accuracy of a recombinant LigA protein based IgM ELISA during outbreaks in the clinical-setting of a highly endemic country. Methodology/Principal Findings A prospective study was conducted from October 2011 to September 2013 at a national referral hospital for infectious diseases in Manila, Philippines. Patients who were hospitalized with clinically suspected leptospirosis were enrolled. Plasma and urine were collected on admission and/or at discharge and tested using the LigA-IgM ELISA and a whole cell-based IgM ELISA. Sensitivity and specificity of these tests were evaluated with cases diagnosed by microscopic agglutination test (MAT), culture and LAMP as the composite reference standard and blood bank donors as healthy controls: the mean+3 standard deviation optical density value of healthy controls was used as the cut-off limit (0.062 for the LigA-IgM ELISA and 0.691 for the whole cell-based IgM ELISA). Of 304 patients enrolled in the study, 270 (89.1%) were male and the median age was 30.5 years; 167 (54.9%) were laboratory confirmed. The sensitivity and ROC curve AUC for the LigA-IgM ELISA was significantly greater than the whole cell-based IgM ELISA (69.5% vs. 54.3%, p<0.01; 0.90 vs. 0.82, p<0.01) on admission, but not at discharge. The specificity of LigA-IgM ELISA and whole cell-based IgM ELISA were not significantly different (98% vs. 97%). Among 158 MAT negative patients, 53 and 28 were positive by LigA- and whole cell-based IgM ELISA, respectively; if the laboratory confirmation was re-defined by LigA-IgM ELISA and LAMP, the clinical findings were more characteristic of leptospirosis than the diagnosis based on MAT/culture/LAMP. Conclusions/Significance The newly

  5. Evaluation of fully automated assays for the detection of Rubella IgM and IgG antibodies by the Elecsys(®) immunoassay system.

    PubMed

    van Helden, Josef; Grangeot-Keros, Liliane; Vauloup-Fellous, Christelle; Vleminckx, Renaud; Masset, Frédéric; Revello, Maria-Grazia

    2014-04-01

    Screening for acute rubella infection in pregnancy is an important element of antenatal care. This study compared the sensitivity, specificity and reproducibility of two new, fully automated Elecsys(®) Rubella IgM and IgG immunoassays designed for the Elecsys 2010, Modular Analytics E170, COBAS e-411 and COBAS e-601 and e602 analytical platforms, with current assays using serum from patients with primary rubella infections, vaccinated patients, patients with potentially cross-reacting infections and on routine samples in clinical laboratories in France, Germany and Italy. Both assays showed good within-run and within-laboratory precision. A sensitivity of 79.8-96.0% was demonstrated for Elecsys IgM in primary, early acute infection, consistent with existing assays. In samples obtained from routine antenatal screening, the Elecsys Rubella IgM assay revealed high specificity (98.7-99.0%). A significantly (p<0.0001) lower reactivity was demonstrated in samples from previously infected patients where acute rubella infection was excluded, and the incidence of false positives in patients with potentially cross-reacting infections was lower with Elecsys Rubella IgM compared with other. The Elecsys Rubella IgG assay exhibited a relative sensitivity of 99.9-100.0% and specificity of 97.4-100.0% in samples from routine antenatal screening. The Elecsys Rubella IgM and IgG assays allow convenient, rapid and reliable determination of anti-rubella antibodies. Sensitivity, specificity and reproducibility were comparable with existing assay systems. Assay results were available in approximately half the time required for currently employed methods and the assays are compatible with widely used analytical platforms.

  6. In Vivo-Expressed Proteins of Virulent Leptospira interrogans Serovar Autumnalis N2 Elicit Strong IgM Responses of Value in Conclusive Diagnosis

    PubMed Central

    Raja, Veerapandian; Shanmughapriya, Santhanam; Kanagavel, Murugesan; Artiushin, Sergey C.; Velineni, Sridhar; Timoney, John F.

    2015-01-01

    Leptospirosis is a serious zoonosis that is underdiagnosed because of limited access to laboratory facilities in Southeast Asia, Central and South America, and Oceania. Timely diagnosis of locally distributed serovars of high virulence is crucial for successful care and outbreak management. Using pooled patient sera, an expression gene library of a virulent Leptospira interrogans serovar Autumnalis strain N2 isolated in South India was screened. The identified genes were characterized, and the purified recombinant proteins were used as antigens in IgM enzyme-linked immunosorbent assay (ELISA) either singly or in combination. Sera (n = 118) from cases of acute leptospirosis along with sera (n = 58) from healthy subjects were tested for reactivity with the identified proteins in an ELISA designed to detect specific IgM responses. We have identified nine immunoreactive proteins, ArgC, RecA, GlpF, FliD, TrmD, RplS, RnhB, Lp28.6, and Lrr44.9, which were found to be highly conserved among pathogenic leptospires. Apparently, the proteins ArgC, RecA, GlpF, FliD, TrmD, and Lrr44.9 are expressed during natural infection of the host and undetectable in in vitro cultures. Among all the recombinant proteins used as antigens in IgM ELISA, ArgC had the highest sensitivity and specificity, 89.8% and 95.5%, respectively, for the conclusive diagnosis of leptospirosis. The use of ArgC and RecA in combination for IgM ELISA increased the sensitivity and specificity to 95.7% and 94.9%, respectively. ArgC and RecA thus elicited specific IgM responses and were therefore effective in laboratory confirmation of Leptospira infection. PMID:26607308

  7. Atheroprotective natural anti-phosphorylcholine antibodies of IgM subclass are decreased in Swedish controls as compared to non-westernized individuals from New Guinea

    PubMed Central

    Frostegård, Johan; Tao, WenJing; Georgiades, Anastasia; Råstam, Lennart; Lindblad, Ulf; Lindeberg, Staffan

    2007-01-01

    Objective To determine the importance of IgM antibodies against phosphorylcholine (aPC), a novel protective factor for cardiovascular disease (CVD), in a population with a non-western life style as compared with a Swedish control group. Methods and results Risk factors for cardiovascular disease were determined in a group of 108 individuals aged 40–86 years from New Guinea and 108 age-and sex-matched individuals from a population based study in Sweden. Antibodies were tested by ELISA. aPC IgM levels were significantly higher among New Guineans than among Swedish controls (p < 0.0001). This difference remained significant among both men and women when controlled for LDL and blood pressure which were lower and smoking which was more prevalent in New Guineans as compared to Swedish controls (p < 0.0001). aPC IgM was significantly and negatively associated with age and systolic blood pressure among Swedish controls and with waist circumference among New Guineans. aPC IgM levels were significantly higher among women than men in both groups. The proportion of the saturated fatty acid (FA) myristic acid in serum cholesterol esters was negatively but polyunsaturated eicosapentaenoic acid and also lipoprotein (a) were positively associated with aPC IgM levels. Conclusion IgM-antibodies against PC, which have atheroprotective properties, are higher in a population from Kitava, New Guinea with a traditional lifestyle, than in Swedish Controls, and higher among women than men in both populations tested. Such antibodies could contribute to the low incidence of cardiovascular disease reported from Kitava and could also provide an explanation as to why women have a later onset of CVD than men. PMID:17374168

  8. Utility of IgM ELISA, TaqMan real-time PCR, reverse transcription PCR, and RT-LAMP assay for the diagnosis of Chikungunya fever.

    PubMed

    Reddy, Vijayalakshmi; Ravi, Vasanthapuram; Desai, Anita; Parida, Manmohan; Powers, Ann M; Johnson, Barbara W

    2012-11-01

    Chikungunya fever a re-emerging infection with expanding geographical boundaries, can mimic symptoms of other infections like dengue, malaria which makes the definitive diagnosis of the infection important. The present study compares the utility of four laboratory diagnostic methods viz. IgM capture ELISA, an in house reverse transcription PCR for the diagnosis of Chikungunya fever, TaqMan real-time PCR, and a one step reverse transcription-loop mediated isothermal amplification assay (RT-LAMP). Out of the 70 serum samples tested, 29 (41%) were positive for Chikungunya IgM antibody by ELISA and 50 (71%) samples were positive by one of the three molecular assays. CHIKV specific nucleic acid was detected in 33/70 (47%) by reverse transcription PCR, 46/70 (66%) by TaqMan real-time PCR, and 43/70 (62%) by RT-LAMP assay. A majority of the samples (62/70; 89%) were positive by at least one of the four assays used in the study. The molecular assays were more sensitive for diagnosis in the early stages of illness (2-5 days post onset) when antibodies were not detectable. In the later stages of illness, the IgM ELISA is a more sensitive diagnostic test. In conclusion we recommend that the IgM ELISA be used as an initial screening test followed one of the molecular assays in samples that are collected in the early phase of illness and negative for CHIKV IgM antibodies. Such as approach would enable rapid confirmation of the diagnosis and implementation of public health measures especially during outbreaks.

  9. IgG avidity assay: a tool for excluding acute toxoplasmosis in prolonged IgM titer sera from pregnant women.

    PubMed

    Emelia, O; Rahana, A R; Mohamad Firdaus, A; Cheng, H S; Nursyairah, M S; Fatinah, A S; Azmawati, M N; Siti, N A M; Aisah, M Y

    2014-12-01

    An accurate diagnosis for toxoplasmosis is crucial for pregnant women as this infection may lead to severe sequelae in the fetus. The value of IgG avidity assay as a tool to determine acute and chronic toxoplasmosis during pregnancy was evaluated in Universiti Kebangsaan Malaysia Medical Centre (UKMMC). In this study, 281 serum samples from 281 pregnant women in various trimesters were collected. These samples were assayed using specific anti-Toxoplasma IgM and IgG antibodies, followed by IgG avidity test. The overall seroprevalence of toxoplasmosis in pregnant women was 35.2% (33.5% for anti-Toxoplasma IgG and 1.8% for both anti-Toxoplasma IgG and IgM antibodies). Of 5 (1.8%) serum samples positive for IgM ELISA, 4 had high-avidity antibodies, suggesting past infection and one sample with borderline avidity index. Two samples with low avidity were from IgM negative serum samples. The IgG avidity assay exhibited an excellent specificity of 97.6% and a negative predictive value (NPV) of 95.6%. The study also demonstrated no significant correlation between avidity indexes of the sera with IgG (r=0.12, p=0.24) and IgM (r=-0.00, p=0.98), suggesting the complementary needs of the two tests for a better diagnosis outcome. These findings highlight the usefulness of IgG avidity assay in excluding a recently acquired toxoplasmosis infection in IgM-positive serum sample.

  10. The importance of quaternary structure in the expression of the C1-binding site of IgM.

    PubMed Central

    Bubb, M O; Conradie, J D

    1976-01-01

    The ability of C1 to bind to the Fc5mu-fragment of a monoclonal IgM was determined by means of a haemolytic C1-inhibition assay. Fc5mu fragments were produced by trypsin digestion at five different temperatures ranging from 54-62 degrees (2 degrees increments) and were purified by immunoadsorption through a column of monospecific anti-Fabmu and by molecular exclusion chromatography. Analytical ultracentrifugation showed the final preparations to be free of aggregates. A plot of mug Fc5mu required to inhibit 50% of available C1 versus temperatures of production of the fragment yielded a curve with a minimum at 58-60 degrees. Upon mild reduction and alkylation of these Fc5mu fragments their C1-fixing capacity became approximately the same irrespective of temperature of production. Fc5mu was also prepared at 25 degrees in the presence of 5 M urea, purified by immunoadsorption as before and aliquots then exposed to temperatures ranging from 40-70 degrees (5 degrees increments) for 15 min. After aggregates had been removed by chromatography a similar minimum in C1-fixation was again observed at 60 degrees. Reduction and alkylation once more abolished these differences. Fc5mu and its reduced and alkylated subunits, produced at 60 degrees and then exposed to various concentrations of urea (0-7 M) for 24 hd did not yield a minimum in C1 fixation. Reduced and alkylated Fcmu incubated at various temperatures (40-70 degrees) also did not fix C1 differentially. Examination in the near and far u.v. region of the circular dichroism spectra of different Fc5mu preparations showed a gradual loss of structure associated with restricted aromatic chromophores and secondary (beta) structure with increased temperature. Urea denaturation had a more pronounced and irreversible effect on Fc5 mu conformation. These changes could not be correlated with the CU-fixation patterns observed. It would therefore appear that elevated temperatures induce a static change in the pentameric FC-part of

  11. Pre-cut Filter Paper for Detecting Anti-Japanese Encephalitis Virus IgM from Dried Cerebrospinal Fluid Spots

    PubMed Central

    Bharucha, Tehmina; Chanthongthip, Anisone; Phuangpanom, Soumphou; Phonemixay, Ooyanong; Sengvilaipaseuth, Onanong; Vongsouvath, Manivanh; Lee, Sue; Newton, Paul N.; Dubot-Pérès, Audrey

    2016-01-01

    Background The use of filter paper as a simple, inexpensive tool for storage and transportation of blood, ‘Dried Blood Spots’ or Guthrie cards, for diagnostic assays is well-established. In contrast, there are a paucity of diagnostic evaluations of dried cerebrospinal fluid (CSF) spots. These have potential applications in low-resource settings, such as Laos, where laboratory facilities for central nervous system (CNS) diagnostics are only available in Vientiane. In Laos, a major cause of CNS infection is Japanese encephalitis virus (JEV). We aimed to develop a dried CSF spot protocol and to evaluate its diagnostic performance using the World Health Organisation recommended anti-JEV IgM antibody capture enzyme-linked immunosorbent assay (JEV MAC-ELISA). Methodology and Principal Findings Sample volumes, spotting techniques and filter paper type were evaluated using a CSF-substitute of anti-JEV IgM positive serum diluted in Phosphate Buffer Solution (PBS) to end-limits of detection by JEV MAC-ELISA. A conventional protocol, involving eluting one paper punch in 200μl PBS, did not detect the end-dilution, nor did multiple punches utilising diverse spotting techniques. However, pre-cut filter paper enabled saturation with five times the volume of CSF-substitute, sufficiently improving sensitivity to detect the end-dilution. The diagnostic accuracy of this optimised protocol was compared with routine, neat CSF in a pilot, retrospective study of JEV MAC-ELISA on consecutive CSF samples, collected 2009–15, from three Lao hospitals. In comparison to neat CSF, 132 CSF samples stored as dried CSF spots for one month at 25–30°C showed 81.6% (65.7–92.3 95%CI) positive agreement, 96.8% (91.0–99.3 95%CI) negative agreement, with a kappa coefficient of 0.81 (0.70–0.92 95%CI). Conclusions/Significance The novel design of pre-cut filter paper saturated with CSF could provide a useful tool for JEV diagnostics in settings with limited laboratory access. It has the

  12. [Significance of IgM titration by an immunoenzyme technic for the serodiagnosis and epidemiological surveillance of dengue in French Polynesia].

    PubMed

    Chungue, E; Boutin, J P; Roux, J

    1989-01-01

    The usefulness of IgM antibody capture enzyme-linked immunosorbent assay (ELISA-IgM) for the diagnosis of dengue was studied using several groups of sera. An attempt was made to detect IgM against each dengue serotype. Of 76 paired sera from patients with dengue type 4 infection, the diagnosis was confirmed by ELISA-IgM in 74: in 47 out of 49 primary dengue infections and in all 27 secondary dengue infections. It was possible to detect IgM in 15 of the acute sera of primary infections and in 16 of the acute sera of secondary infections. In 20 patients from whom only single serum with high haemagglutination-inhibition titre (greater than or equal to 640) was available, 18 were found positive for dengue IgM. Dengue infection was also confirmed in 10 patients with dengue type 1 and in 10 patients with dengue type 2. IgM was detected in 1 of the acute sera of patients with either dengue type 1 or dengue type 2 infection. In all cases in which IgM was detected, a positive response was obtained for the homologous antigen. ELISA-IgM on sequential serum samples from 7 proven cases of dengue type 4 revealed that antibody titres greater than or equal to 400 are reached early in the second week of the illness and persist for 60 days or more at a low level (titre less than 400). An attempt was made to obtain a type-specific presumptive diagnosis by analysing the relative titre against each of the 4 antigens. It was possible to make a diagnosis of dengue type 4 in 85% and 52% of IgM-positive responses in primary and secondary infections (p less than 0.01), respectively, using paired sera from patients with known dengue type 4 infection. ELISA-IgM was shown to be of interest in the diagnosis and surveillance of dengue in French Polynesia where dengue viruses are, presently, the only known flavivirus.

  13. Th17 responses and natural IgM antibodies are related to gut microbiota composition in systemic lupus erythematosus patients

    PubMed Central

    López, Patricia; de Paz, Banesa; Rodríguez-Carrio, Javier; Hevia, Arancha; Sánchez, Borja; Margolles, Abelardo; Suárez, Ana

    2016-01-01

    Intestinal dysbiosis, characterized by a reduced Firmicutes/Bacteroidetes ratio, has been reported in systemic lupus erythematosus (SLE) patients. In this study, in vitro cultures revealed that microbiota isolated from SLE patient stool samples (SLE-M) promoted lymphocyte activation and Th17 differentiation from naïve CD4+ lymphocytes to a greater extent than healthy control-microbiota. Enrichment of SLE-M with Treg-inducing bacteria showed that a mixture of two Clostridia strains significantly reduced the Th17/Th1 balance, whereas Bifidobacterium bifidum supplementation prevented CD4+ lymphocyte over-activation, thus supporting a possible therapeutic benefit of probiotics containing Treg-inducer strains in order to restore the Treg/Th17/Th1 imbalance present in SLE. In fact, ex vivo analyses of patient samples showed enlarged Th17 and Foxp3+ IL-17+ populations, suggesting a possible Treg-Th17 trans-differentiation. Moreover, analyses of fecal microbiota revealed a negative correlation between IL-17+ populations and Firmicutes in healthy controls, whereas in SLE this phylum correlated directly with serum levels of IFNγ, a Th1 cytokine slightly reduced in patients. Finally, the frequency of Synergistetes, positively correlated with the Firmicutes/Bacteroidetes ratio in healthy controls, tended to be reduced in patients when anti-dsDNA titers were increased and showed a strong negative correlation with IL-6 serum levels and correlated positively with protective natural IgM antibodies against phosphorylcholine. PMID:27044888

  14. In vitro synthesis of IgM rheumatoid factor in response to Staphylococcus aureus, by lymphocytes from healthy adults

    SciTech Connect

    Goldstein, R.; Karsh, J.

    1986-12-01

    Peripheral blood mononuclear cells from 20 healthy adults were tested in vitro for the production of IgM rheumatoid factor (RF) in response to Staphylococcus aureus Cowan I (SAC) or pokeweed mitogen. Fifteen of the 20 normal subjects produced greater than or equal to 4 ng/ml IgM-RF (mean +/- SD 46 +/- 55 ng/ml) in response to SAC, compared with only 2 of 20 who produced greater than or equal to 4 ng/ml IgM-RF (mean +/- SD 2 +/- 4 ng/ml) in response to pokeweed mitogen (P = 0.0001). Separation and reconstitution of autologous T and B cell-enriched fractions, with and without prior T cell irradiation, provided evidence for a radiosensitive T helper/inducer cell involved in the IgM-RF response to SAC in 70% of the normal subjects studied. SAC appears to be a potent stimulus of IgM-RF production, with a cellular mechanism distinct from that of other in vitro systems.

  15. Generation of Two-color Antigen Microarrays for the Simultaneous Detection of IgG and IgM Autoantibodies.

    PubMed

    Chruscinski, Andrzej; Huang, Flora Y Y; Ulndreaj, Antigona; Chua, Conan; Fehlings, Michael; Rao, Vivek; Ross, Heather J; Levy, Gary A

    2016-01-01

    Autoantibodies, which are antibodies against self-antigens, are present in many disease states and can serve as markers for disease activity. The levels of autoantibodies to specific antigens are typically detected with the enzyme-linked immunosorbent assay (ELISA) technique. However, screening for multiple autoantibodies with ELISA can be time-consuming and requires a large quantity of patient sample. The antigen microarray technique is an alternative method that can be used to screen for autoantibodies in a multiplex fashion. In this technique, antigens are arrayed onto specially coated microscope slides with a robotic microarrayer. The slides are probed with patient serum samples and subsequently fluorescent-labeled secondary antibodies are added to detect binding of serum autoantibodies to the antigens. The autoantibody reactivities are revealed and quantified by scanning the slides with a scanner that can detect fluorescent signals. Here we describe methods to generate custom antigen microarrays. Our current arrays are printed with 9 solid pins and can include up to 162 antigens spotted in duplicate. The arrays can be easily customized by changing the antigens in the source plate that is used by the microarrayer. We have developed a two-color secondary antibody detection scheme that can distinguish IgG and IgM reactivities on the same slide surface. The detection system has been optimized to study binding of human and murine autoantibodies. PMID:27685156

  16. Th17 responses and natural IgM antibodies are related to gut microbiota composition in systemic lupus erythematosus patients.

    PubMed

    López, Patricia; de Paz, Banesa; Rodríguez-Carrio, Javier; Hevia, Arancha; Sánchez, Borja; Margolles, Abelardo; Suárez, Ana

    2016-04-05

    Intestinal dysbiosis, characterized by a reduced Firmicutes/Bacteroidetes ratio, has been reported in systemic lupus erythematosus (SLE) patients. In this study, in vitro cultures revealed that microbiota isolated from SLE patient stool samples (SLE-M) promoted lymphocyte activation and Th17 differentiation from naïve CD4(+) lymphocytes to a greater extent than healthy control-microbiota. Enrichment of SLE-M with Treg-inducing bacteria showed that a mixture of two Clostridia strains significantly reduced the Th17/Th1 balance, whereas Bifidobacterium bifidum supplementation prevented CD4(+) lymphocyte over-activation, thus supporting a possible therapeutic benefit of probiotics containing Treg-inducer strains in order to restore the Treg/Th17/Th1 imbalance present in SLE. In fact, ex vivo analyses of patient samples showed enlarged Th17 and Foxp3(+) IL-17(+) populations, suggesting a possible Treg-Th17 trans-differentiation. Moreover, analyses of fecal microbiota revealed a negative correlation between IL-17(+) populations and Firmicutes in healthy controls, whereas in SLE this phylum correlated directly with serum levels of IFNγ, a Th1 cytokine slightly reduced in patients. Finally, the frequency of Synergistetes, positively correlated with the Firmicutes/Bacteroidetes ratio in healthy controls, tended to be reduced in patients when anti-dsDNA titers were increased and showed a strong negative correlation with IL-6 serum levels and correlated positively with protective natural IgM antibodies against phosphorylcholine.

  17. Syndrome of selective IgM deficiency with severe T cell deficiency associated with disseminated cutaneous mycobacterium avium intracellulaire infection

    PubMed Central

    Gharib, Asal; Louis, Ankmalika Gupta; Agrawal, Sudhanshu; Gupta, Sudhir

    2015-01-01

    Cutaneous non-disseminated, non-tuberculous mycobacterial infections have been reported in both immunocompetent and immunocompromised subjects. Systemic Mycobacterium avium intracellulaire (MAI) have been reported in non-HIV patients with Idiopathic CD4 lymphocytopenia. We report a comprehensive immunological analysis in syndrome of selective IgM deficiency and T lymphocytopenia (both CD4+ and CD8+) with disseminated cutaneous MAI infection. Naïve (TN) and Central memory (TCM) subsets of both CD4+ and CD8+ T cells were decreased, whereas terminally differentiated effector memory (TEMRA) subset of CD4+ and CD8+ T cells were markedly increased. IFN-γ producing T cells were markedly decreased. Although CD14highCD16- proinflammatory monocytes were modestly increased, IFN-γR+ monocytes were markedly decreased. The expression of TLR3, TLR5, TLR7, and TLR9 on monocytes was decreased. Germinal center B cells (CD19+IgD-CD38+CD27lo) and B1 cells (CD20+CD27+CD43+CD70-) were markedly decreased. A role of immune alterations, including B cells and antibodies in disseminated cutaneous MAI infection is discussed. PMID:26550546

  18. Motor-dominant polyneuropathy due to IgM monoclonal antibody against disialosyl gangliosides in a patient with mantle cell lymphoma.

    PubMed

    Mori, Akio; Ueno, Yuji; Kuroki, Takuma; Hoshino, Yasunobu; Shimura, Hideki; Sekiguchi, Yasunobu; Noguchi, Masaaki; Hamada, Yukihiro; Kusunoki, Susumu; Hattori, Nobutaka; Urabe, Takao

    2014-02-15

    A rapidly progressive motor-dominant neuropathy associated with IgM monoclonal antibody against gangliosides with disialosyl residues, GD3, GD1b, GT1b, and GQ1b, in a 60-year-old Japanese man with mantle cell lymphoma is reported. Plasma exchange and chemotherapy for mantle cell lymphoma were performed for the neuropathy and mantle cell lymphoma. After therapy, the motor neuropathy dramatically improved concurrently with substantial reduction of the antibody activities especially in reaction to GD1b. This is the first case report of neuropathy with anti-disialosyl IgM antibodies associated with mantle cell lymphoma, and plasma exchange and chemotherapy were effective. PMID:24315580

  19. Al Composites

    NASA Astrophysics Data System (ADS)

    Chandanayaka, Tharaka; Azarmi, Fardad

    2014-05-01

    In the present study, cold spraying technique was used to fabricate a metal matrix composite (MMC) that consists of Ni matrix and 20 vol.% Ni3Al particles at two different particle sizes as reinforcement. This study intends to investigate the effect of reinforcement particle size on microstructural and mechanical properties of cold sprayed MMCs. Two different Ni3Al powders with nominal particle size of -45 to +5 and +45 to 100 μm were used as reinforcement in this study. Cold sprayed Ni-Ni3Al samples were subjected to the microstructural observation and characterization prior to any mechanical testing. Then, samples were tested using nano-indentation, Knoop hardness, Vickers hardness, and Resonance frequency to evaluate their mechanical properties. No significant changes were observed in microstructural characteristics due to different particle sizes. The results obtained from a variety of mechanical testings indicated that the increasing reinforcement particle size resulted in the slight reduction of mechanical properties such as elastic modulus and hardness in cold sprayed MMCs. The mechanical interlock between deposited particles defines the bonding strength in cold sprayed samples. Small size particles have a higher velocity and impact resulting in stronger interlock between deformed particles.

  20. Enzyme immunoassays (EIAs) for the detection of anti-Haemophilus ducreyi serum IgA, IgG, and IgM antibodies.

    PubMed

    Roggen, E L; Hoofd, G; Van Dyck, E; Piot, P

    1994-01-01

    In Belgium, the Department of Infection and Immunity of the Institute of Tropical Medicine in Antwerp modified an experimental enzyme immunoassay (EIA) for the detection of serum IgG to Hemophilus ducreyi to develop EIAs for detection of anti-H. ducreyi IgA and IgM antibodies. They tested the modified EIA on sera from people in Nairobi, Kenya; Kigali, Rwanda; Banjul, The Gambia; and Bangkok, Thailand, who had a sexually transmitted disease. The EIA was able to identify correctly those who did not have anti-H ducreyi IgA, IgG, and IgM antibodies in 97%, 92%, and 99% of cases, respectively. Among people with a genital ulceration for more than 8 days, it was able to identify correctly those who had IgA, IgG, and IgM antibodies in 88%, 93%, and 78% of cases, respectively. 95% of all culture-proven chancroid patients tested seropositive for at least 1 antibody type. The sensitivity of IgG and IgA EIAs was significantly enhanced in patients with culture-proven chancroid who were older than 24 years old (p .01). HIV seropositive people from Kigali who had culture-proven chancroid had higher anti-H. ducreyi IgG seropositivity rates (but not IgA and IgM seropositivity rates), than did HIV seronegative chancroid people from Kigali (p .05). The increased IgG seropositivity rate was not related to higher antibody titers, however, suggesting that HIV infection modifies the response to H. ducreyi. These results show that the 3 EIAs hold promise as a means to study the kinetics of antibodies and the epidemiology of chancroid.

  1. Biosynthesis of immunoglobulin A (IgA) and immunoglobulin M (IgM). Requirement for J chain and a disulphide-exchanging enzyme for polymerization

    PubMed Central

    Corte, E. Della; Parkhouse, R. M. E.

    1973-01-01

    Mouse myeloma cells secreting 19S IgM (immunoglobulin M) (MOPC 104E and TEPC 183) or monomer and polymer IgA (immunoglobulin A) (MOPC 315) were incubated with radioactive leucine and the intracellular and secreted immunoglobulins and immunoglobulin subunits were prepared by preparative sucrose-density-gradient centrifugation. Samples were reduced in the presence or absence of isolated J chain, passed over Sephadex G-25 and then incubated at 37°C for 30min with or without a source of disulphide-interchange enzyme. The extent of reassembly of reduced subunits was then evaluated by electrophoresis in polyacrylamide gels. Provided that J chain and the disulphide-interchange enzyme were supplied, both IgM and IgA could be assembled from their respective subunits, obtained by reductive cleavage of polymeric forms. Under similar conditions, assembly of polymeric forms from intracellular or secreted 7S monomer subunits also occurred. Under these conditions polymerization was total, there being no residue of the monomeric form. Reassembly did not occur in the absence of either J chain or the enzyme. All of the J chain released from IgM by reductive cleavage was incorporated back into the reassembled polymer. The J chain is therefore likely to be an essential structural requirement for polymeric immunoglobulins. A variety of controls ruled out non-specific interactions, and further suggested that the amino acid sequence of polypeptide chains determines the specificity of polymerization. The fact that intracellular IgA and IgM monomer subunits known to be deficient in galactose and fucose can be completely polymerized suggests that the addition of carbohydrate does not control polymerization. ImagesFig. 7. PMID:4205352

  2. Multiplex Immunoassay for Lyme Disease Using VlsE1-IgG and pepC10-IgM Antibodies: Improving Test Performance through Bioinformatics ▿

    PubMed Central

    Porwancher, Richard B.; Hagerty, C. Greg; Fan, Jianqing; Landsberg, Lisa; Johnson, Barbara J. B.; Kopnitsky, Mark; Steere, Allen C.; Kulas, Karen; Wong, Susan J.

    2011-01-01

    The Centers for Disease Control and Prevention currently recommends a 2-tier serologic approach to Lyme disease laboratory diagnosis, comprised of an initial serum enzyme immunoassay (EIA) for antibody to Borrelia burgdorferi followed by supplementary IgG and IgM Western blotting of EIA-positive or -equivocal samples. Western blot accuracy is limited by subjective interpretation of weakly positive bands, false-positive IgM immunoblots, and low sensitivity for detection of early disease. We developed an objective alternative second-tier immunoassay using a multiplex microsphere system that measures VlsE1-IgG and pepC10-IgM antibodies simultaneously in the same sample. Our study population comprised 79 patients with early acute Lyme disease, 82 patients with early-convalescent-phase disease, 47 patients with stage II and III disease, 34 patients post-antibiotic treatment, and 794 controls. A bioinformatic technique called partial receiver-operator characteristic (ROC) regression was used to combine individual antibody levels into a single diagnostic score with a single cutoff; this technique enhances test performance when a high specificity is required (e.g., ≥95%). Compared to Western blotting, the multiplex assay was equally specific (95.6%) but 20.7% more sensitive for early-convalescent-phase disease (89.0% versus 68.3%, respectively; 95% confidence interval [95% CI] for difference, 12.1% to 30.9%) and 12.5% more sensitive overall (75.0% versus 62.5%, respectively; 95% CI for difference, 8.1% to 17.1%). As a second-tier test, a multiplex assay for VlsE1-IgG and pepC10-IgM antibodies performed as well as or better than Western blotting for Lyme disease diagnosis. Prospective validation studies appear to be warranted. PMID:21367982

  3. Multiplex immunoassay for Lyme disease using VlsE1-IgG and pepC10-IgM antibodies: improving test performance through bioinformatics.

    PubMed

    Porwancher, Richard B; Hagerty, C Greg; Fan, Jianqing; Landsberg, Lisa; Johnson, Barbara J B; Kopnitsky, Mark; Steere, Allen C; Kulas, Karen; Wong, Susan J

    2011-05-01

    The Centers for Disease Control and Prevention currently recommends a 2-tier serologic approach to Lyme disease laboratory diagnosis, comprised of an initial serum enzyme immunoassay (EIA) for antibody to Borrelia burgdorferi followed by supplementary IgG and IgM Western blotting of EIA-positive or -equivocal samples. Western blot accuracy is limited by subjective interpretation of weakly positive bands, false-positive IgM immunoblots, and low sensitivity for detection of early disease. We developed an objective alternative second-tier immunoassay using a multiplex microsphere system that measures VlsE1-IgG and pepC10-IgM antibodies simultaneously in the same sample. Our study population comprised 79 patients with early acute Lyme disease, 82 patients with early-convalescent-phase disease, 47 patients with stage II and III disease, 34 patients post-antibiotic treatment, and 794 controls. A bioinformatic technique called partial receiver-operator characteristic (ROC) regression was used to combine individual antibody levels into a single diagnostic score with a single cutoff; this technique enhances test performance when a high specificity is required (e.g., ≥ 95%). Compared to Western blotting, the multiplex assay was equally specific (95.6%) but 20.7% more sensitive for early-convalescent-phase disease (89.0% versus 68.3%, respectively; 95% confidence interval [95% CI] for difference, 12.1% to 30.9%) and 12.5% more sensitive overall (75.0% versus 62.5%, respectively; 95% CI for difference, 8.1% to 17.1%). As a second-tier test, a multiplex assay for VlsE1-IgG and pepC10-IgM antibodies performed as well as or better than Western blotting for Lyme disease diagnosis. Prospective validation studies appear to be warranted.

  4. Evaluation of end-point titration, single dilution and capture enzyme immunoassays for measurement of antirotaviral IgA and IgM in infantile secretions and serum.

    PubMed

    Coulson, B S; Grimwood, K; Bishop, R F; Barnes, G L

    1989-10-01

    In order to facilitate measurement of antirotaviral IgA in large collections of faeces and secretions, adaptations of enzyme immunoassay methods for estimating antirotaviral IgA and IgM in duodenal fluid, saliva, faeces and serum were studied. To quantitate specific IgA, a single dilution of each sample was assayed. Results were expressed as antirotaviral IgA units derived from a standard curve. Units were calculated by log-logit analysis on computer. There was strong correlation between antirotaviral IgA units and end-point titres in 257 faecal samples (correlation coefficient r = 0.92) and in 182 duodenal fluids and salivary samples (correlation coefficient r = 0.74). The assay was validated using acute and convalescent faeces from children with or without rotavirus infection. Immune conversions in IgA were detected in 33 (75%) of the children by units and 34 (77%) by titres. None of nine children with gastroenteritis due to other infectious agents showed immune conversions to rotavirus. A monoclonal capture IgM assay showed similar end-point titres and numbers of immune conversions when compared with a direct assay for antirotaviral IgM in serum and secretions. Use of the capture method eliminated false-positive reactions with the cell control. The assay for antirotaviral IgA units in secretions is simple, rapid, reproducible and reliable, and has proven of value in longitudinal epidemiological studies of rotavirus coproIgA profiles. Both the capture IgM technique and the single dilution IgA method permit analysis of large numbers of specimens and are appropriate for examination of immune responses to natural rotavirus infection or during vaccine trials.

  5. Molecular cloning of IgZ heavy chain isotype in Catla catla and comparative expression profile of IgZ and IgM following pathogenic infection.

    PubMed

    Patel, Bhakti; Banerjee, Rajanya; Basu, Madhubanti; Lenka, Saswati; Samanta, Mrinal; Das, Surajit

    2016-08-01

    Immunoglobulins serve as a crucial arm of the adaptive immune system against detrimental pathogenic threats in teleosts. However, whether the novel Ig isotype IgZ is present in the Indian major carp, Catla catla, has not yet been elucidated. The present study reports the presence of IgZ ortholog in C. catla (CcIgZ) and further demonstrates its comparative tissue specific expression with IgM (CcIgM) in response to bacterial and parasitic stimulation. The putative 139 amino acid sequence of IgZ heavy chain cDNA of C. catla showed homology with IgZ constant domains of other teleosts. Phylogenetic analysis of the predicted IgZ transcript sequence clustered with previously identified IgZ heavy chain sequences of Cyprinidae family members. The inductive expression profiles of IgZ and IgM genes were evaluated in immunologically relevant tissues at 24, 48 and 72 hr post infection with Aeromonas hydrophila, Streptococcus uberis and Argulus sp. Both CcIgZ and CcIgM were expressed most strongly in the kidneys of healthy fish. Basal expression of CcIgM transcript was higher than that of CcIgZ in all the examined tissues. Stimulation with bacteria triggered significant increase of IgZ in the intestine (P < 0.001) and spleen (P < 0.01), whereas IgM was relatively up-regulated in blood (P < 0.001) after stimulation with each of the three pathogens assessed. The study is the first to report identification of IgZ in C. catla. Further, it provides insights into the differential expression profiles of IgZ and IgM isotypes against various pathogenic infection in C. catla, which may facilitate better prophylaxis again such infections. PMID:27301776

  6. Antigenic modulation of the cytophilic binding of guinea-pig IgG and IgM antibodies to homologous macrophages.

    PubMed Central

    Webster, R O; Lawrence, D A

    1979-01-01

    The cytophilic binding of immune complexes by peritoneal exudate cells (PEC) from adjuvant-stimulated guinea-pigs was studied using 125I-labelled guinea-pig IgG1, IgG2 and IgM antibodies to the dinitrophenyl (DNP) group. The influence of hapten density upon cytophilic activity was studied by the addition of DNP-conjugated antigens to antibody in 2-200 molar ratios of DNP:antibody. Only IgG2 binding was enhanced by immune complex formation, and the increased binding of IgG2 anti-DNP was dependent on the number of DNP determinants per antigen molecule. Cytophilic activity with epsilon-DNP-L-lysine (DNP-LYS), alpha,epsilon-di-DNP-L-lysine (DNP-LYS-DNP), or DNP1-8-BSA was no greater than that seen in the absence of hapten. Increased cytophilic binding was noted only with DNP20-41-BSA. The binding of IgG2 and IgG2 anti-DNP:DNP-bovine serum albumin (BSA) complexes was inhibited by monomeric IgG2. The relative cytophilic capacities of guinea-pig immunoglobulins appeared as follows: IgG greater than IgG1 greater than IgM. IgG1 and IgM binding of DNP conjugates did not enhance their cytophilic activity; therefore, IgG1 and IgM cytophilic binding to PEC was considered biologically insignificant. This investigation provides further evidence that cytophilic binding of immune complexes to macrophages is due to the co-operative action of multiple Fc sites rather than a conformational change in the IgG2 antibodies, and serum proteins, notably complement components, can alter the binding and/or phagocytosis of IgG2 anti-DNP:DNP-BSA complexes. PMID:86509

  7. Detection of Human Papillomavirus 16-Specific IgG and IgM Antibodies in Patient Sera: A Potential Indicator of Oral Squamous Cell Carcinoma Risk Factor

    PubMed Central

    Kerishnan, Jesinda P.; Gopinath, Subash C.B.; Kai, Sia Bik; Tang, Thean-Hock; Ng, Helen Lee-Ching; Rahman, Zainal Ariff Abdul; Hashim, Uda; Chen, Yeng

    2016-01-01

    The association between human papillomavirus type 16 (HPV16) and oral cancer has been widely reported. However, detecting anti-HPV antibodies in patient sera to determine risk for oral squamous cell carcinoma (OSCC) has not been well studied. In the present investigation, a total of 206 OSCC serum samples from the Malaysian Oral Cancer Database & Tissue Bank System, with 134 control serum samples, were analyzed by enzyme-linked immunosorbant assay (ELISA) to detect HPV16-specific IgG and IgM antibodies. In addition, nested PCR analysis using comprehensive consensus primers (PGMY09/11 and GP5+/6+) was used to confirm the presence of HPV. Furthermore, we have evaluated the association of various additional causal factors (e.g., smoking, alcohol consumption, and betel quid chewing) in HPV-infected OSCC patients. Statistical analysis of the Malaysian population indicated that OSCC was more prevalent in female Indian patients that practices betel quid chewing. ELISA revealed that HPV16 IgG, which demonstrates past exposure, could be detected in 197 (95.6%) OSCC patients and HPV16-specific IgM was found in a total of 42 (20.4%) OSCC patients, indicating current exposure. Taken together, our study suggest that HPV infection may play a significant role in OSCC (OR: 13.6; 95% CI: 3.89-47.51) and HPV16-specific IgG and IgM antibodies could represent a significant indicator of risk factors in OSCC patients. PMID:27279791

  8. Detection of Human Papillomavirus 16-Specific IgG and IgM Antibodies in Patient Sera: A Potential Indicator of Oral Squamous Cell Carcinoma Risk Factor.

    PubMed

    Kerishnan, Jesinda P; Gopinath, Subash C B; Kai, Sia Bik; Tang, Thean-Hock; Ng, Helen Lee-Ching; Rahman, Zainal Ariff Abdul; Hashim, Uda; Chen, Yeng

    2016-01-01

    The association between human papillomavirus type 16 (HPV16) and oral cancer has been widely reported. However, detecting anti-HPV antibodies in patient sera to determine risk for oral squamous cell carcinoma (OSCC) has not been well studied. In the present investigation, a total of 206 OSCC serum samples from the Malaysian Oral Cancer Database & Tissue Bank System, with 134 control serum samples, were analyzed by enzyme-linked immunosorbant assay (ELISA) to detect HPV16-specific IgG and IgM antibodies. In addition, nested PCR analysis using comprehensive consensus primers (PGMY09/11 and GP5(+)/6(+)) was used to confirm the presence of HPV. Furthermore, we have evaluated the association of various additional causal factors (e.g., smoking, alcohol consumption, and betel quid chewing) in HPV-infected OSCC patients. Statistical analysis of the Malaysian population indicated that OSCC was more prevalent in female Indian patients that practices betel quid chewing. ELISA revealed that HPV16 IgG, which demonstrates past exposure, could be detected in 197 (95.6%) OSCC patients and HPV16-specific IgM was found in a total of 42 (20.4%) OSCC patients, indicating current exposure. Taken together, our study suggest that HPV infection may play a significant role in OSCC (OR: 13.6; 95% CI: 3.89-47.51) and HPV16-specific IgG and IgM antibodies could represent a significant indicator of risk factors in OSCC patients.

  9. Complement activation by necrotic cells in normal plasma environment compares to that by late apoptotic cells and involves predominantly IgM.

    PubMed

    Ciurana, Caroline L F; Zwart, Bas; van Mierlo, Gerard; Hack, C Erik

    2004-09-01

    Necrotic cells are generally considered to stimulate inflammation, whereas apoptotic cells should not. However, apoptotic cells have pro-inflammatory properties since they can activate complement. To what extent this activation compares to that by necrotic cells is not known. We compared complement activation by necrotic cells and apoptotic cells in plasma. Jurkat cells were made apoptotic or necrotic by incubation with etoposide or by heat shock, respectively. Cells incubated in recalcified plasma were tested for C3 and C4 fixation and fluid phase generation of complement activation products. Fixation of C3 and C4 to necrotic cells occurred mainly via the classical pathway, independent from the method of necrosis induction and the cell type. Depletion of IgM from plasma almost completely abrogated complement fixation by necrotic cells, which was restored by supplementation with purified IgM. Complement activation by late apoptotic cells was comparable to that by necrotic cells regarding the extent and dependence on IgM. Moreover, incubation of plasma with necrotic or late apoptotic cells led to the generation of comparable amounts of complement activation products. These results indicate that late apoptotic and necrotic cells employ similar complement activation mechanisms in the plasma environment.

  10. Heterozygous Mutation in IκBNS Leads to Reduced Levels of Natural IgM Antibodies and Impaired Responses to T-Independent Type 2 Antigens

    PubMed Central

    Pedersen, Gabriel K.; Ádori, Monika; Stark, Julian M.; Khoenkhoen, Sharesta; Arnold, Carrie; Beutler, Bruce; Karlsson Hedestam, Gunilla B.

    2016-01-01

    Mice deficient in central components of classical NF-κB signaling have low levels of circulating natural IgM antibodies and fail to respond to immunization with T-independent type 2 (TI-2) antigens. A plausible explanation for these defects is the severely reduced numbers of B-1 and marginal zone B (MZB) cells in such mice. By using an ethyl-N-nitrosourea mutagenesis screen, we identified a role for the atypical IκB protein IκBNS in humoral immunity. IκBNS-deficient mice lack B-1 cells and have severely reduced numbers of MZB cells, and thus resemble several other strains with defects in classical NF-κB signaling. We analyzed mice heterozygous for the identified IκBNS mutation and demonstrate that these mice have an intermediary phenotype in terms of levels of circulating IgM antibodies and responses to TI-2 antigens. However, in contrast to mice that are homozygous for the IκBNS mutation, the heterozygous mice had normal frequencies of B-1 and MZB cells. These results suggest that there is a requirement for IκBNS expression from two functional alleles for maintaining normal levels of circulating natural IgM antibodies and responses to TI-2 antigens. PMID:26973645

  11. Detection of IgM Antibrucella Antibody in the Absence of IgGs: A Challenge for the Clinical Interpretation of Brucella Serology

    PubMed Central

    Solís García del Pozo, Julián; Lorente Ortuño, Santiago; Navarro, Elena; Solera, Javier

    2014-01-01

    The use of enzyme-linked immunosorbent assay (ELISA) for the detection of IgG and IgM antibodies antibrucella has become widespread in the diagnosis of human brucellosis. IgM anti-Brucella antibodies are indicative of acute infection. Between 2009–2013, 5307 patients were evaluated for serologic diagnosis at the Microbiology Laboratory of the Albacete General Hospital. A ELISA IgM-positive, IgG-negative anti-Brucella antibody serology pattern was detected in 17 of those patients. Epidemiology data, symptoms, laboratory data, treatment and outcome from these patients were reviewed. Sixteen patients presented with musculoskeletal pain, fatigue and/or fever and 1 was asymptomatic. Five patients received treatment with doxycycline combined with rifampin, gentamycin or streptomycin during 6–12 weeks, with no improvement. None of the 17 patients were finally diagnosed with brucellosis. Our results indicate that anti-Brucella IgM positive serology, per se, is not enough to diagnose acute brucellosis and other methods should be used for confirmation. Brucella serology data should be interpreted taking into account the patient's clinical history and epidemiological context. PMID:25474572

  12. Heterozygous Mutation in IκBNS Leads to Reduced Levels of Natural IgM Antibodies and Impaired Responses to T-Independent Type 2 Antigens.

    PubMed

    Pedersen, Gabriel K; Ádori, Monika; Stark, Julian M; Khoenkhoen, Sharesta; Arnold, Carrie; Beutler, Bruce; Karlsson Hedestam, Gunilla B

    2016-01-01

    Mice deficient in central components of classical NF-κB signaling have low levels of circulating natural IgM antibodies and fail to respond to immunization with T-independent type 2 (TI-2) antigens. A plausible explanation for these defects is the severely reduced numbers of B-1 and marginal zone B (MZB) cells in such mice. By using an ethyl-N-nitrosourea mutagenesis screen, we identified a role for the atypical IκB protein IκBNS in humoral immunity. IκBNS-deficient mice lack B-1 cells and have severely reduced numbers of MZB cells, and thus resemble several other strains with defects in classical NF-κB signaling. We analyzed mice heterozygous for the identified IκBNS mutation and demonstrate that these mice have an intermediary phenotype in terms of levels of circulating IgM antibodies and responses to TI-2 antigens. However, in contrast to mice that are homozygous for the IκBNS mutation, the heterozygous mice had normal frequencies of B-1 and MZB cells. These results suggest that there is a requirement for IκBNS expression from two functional alleles for maintaining normal levels of circulating natural IgM antibodies and responses to TI-2 antigens.

  13. Gold magnetic nanoparticle conjugate-based lateral flow assay for the detection of IgM class antibodies related to TORCH infections.

    PubMed

    Li, Xingxing; Zhang, Qinlu; Hou, Peng; Chen, Mingwei; Hui, Wenli; Vermorken, Alphons; Luo, Zhiyi; Li, Hong; Li, Qin; Cui, Yali

    2015-11-01

    In this study, a lateral flow immunochromatographic assay (LFIA) system for the detection of immunoglobulin M (IgM) antibodies, related to TORCH [(T)oxoplasmosis, (O)ther agents, (R)ubella (also known as German Measles), (C)ytomegalovirus, and (H)erpes simplex virus infections], based on gold magnetic nanoparticles, was established. Following modification with poly(methacrylic acid), the gold magnetic nanoparticles conjugated with an anti‑human IgM antibody (μ‑chain specific) to construct a probe. A lateral flow assay device was constructed based on these conjugates. IgM antibodies to four types of pathogens, notably toxoplasmosis, rubella virus, cytomegalovirus and herpes simplex virus type 2, were detected using this device. Compared with commercial colloidal gold‑based LFIA strips, our method exhibited higher sensitivity. No interference with triglycerides, hemoglobin and bilirubin occurred, and no cross‑reactivity was noted among the four pathogens. The gold magnetic nanoparticle‑LFIA strips were used to assess 41 seropositive and 121 seronegative serum samples. The sensitivity was 100% (162/162) and the specificity was 100% (162/162). This method cannot only be used for the detection of TORCH IgM-specific antibodies, but it can potentially be developed for use in the diagnosis of other acute or recently identified autoimmune diseases. PMID:26329478

  14. Aberrant levels of natural IgM antibodies in osteoarthritis and rheumatoid arthritis patients in comparison to healthy controls.

    PubMed

    Nguyen, Tue G; McKelvey, Kelly J; March, Lyn M; Hunter, David J; Xue, Meilang; Jackson, Christopher J; Morris, Jonathan M

    2016-02-01

    Natural IgM antibodies (nIgM) are polyreactive autoantibodies that have diverse roles in regulating autoimmunity, systemic inflammation and removal of oxidized low-density lipoproteins (oxLDL). We hypothesized that aberrant states of nIgM may exist in persons with osteoarthritis (OA) and rheumatoid arthritis (RA). Herein, we characterized and compared the levels of nIgM specific for phosphorylcholine (anti-PC), double-stranded DNA (anti-dsDNA), and galactosyl (anti-Gal) in persons with OA, RA and healthy controls (HC). Levels of anti-PC nIgM in OA patients were significantly lower than both HC and RA patients in an age-adjusted analysis (P<0.05). In contrast, anti-Gal nIgM levels were significantly higher in RA patients than OA patients (P<0.05) and markedly increased in comparison to HC. Anti-PC nIgM significantly correlated with anti-dsDNA and anti-Gal nIgM levels in HC and RA (P<0.05) but not in OA patients. Elevated CRP levels were associated with RA conditions and old ages in general. There was no significant correlation between anti-PC nIgM and CRP or oxLDL levels. Our study highlights for the first time the evidence of aberrant state of nIgM in human OA compared to healthy individuals that implicates a deficiency in immune responses to oxLDL which may contribute to the metabolic syndromes in the development of OA.

  15. Downstream class switching leads to IgE antibody production by B lymphocytes lacking IgM switch regions.

    PubMed

    Zhang, Tingting; Franklin, Andrew; Boboila, Cristian; McQuay, Amy; Gallagher, Michael P; Manis, John P; Khamlichi, Ahmed Amine; Alt, Frederick W

    2010-02-16

    Ig heavy chain (IgH) class-switch recombination (CSR) replaces the IgH C mu constant region exons with one of several sets of downstream IgH constant region exons (e.g., C gamma, C epsilon, or C alpha), which affects switching from IgM to another IgH class (e.g., IgG, IgE, or IgA). Activation-induced cytidine deaminase (AID) initiates CSR by promoting DNA double-strand breaks (DSBs) within switch (S) regions flanking the donor C mu (S mu) and a downstream acceptor C(H) (e.g., S gamma, S epsilon, S alpha) that are then joined to complete CSR. DSBs generated in S mu frequently are joined within S mu to form internal switch region deletions (ISD). AID-induced ISD and mutations have been considered rare in downstream S regions, suggesting that AID targeting to these S regions requires its prior recruitment to S mu. We have now assayed for CSR and ISD in B cells lacking S mu (S mu(-/-) B cells). In S mu(-/-) B cells activated for CSR to IgG1 and IgE, CSR to IgG1 was greatly reduced; but, surprisingly, CSR to IgE occurred at nearly normal levels. Moreover, normal B cells had substantial S gamma1 ISD and increased mutations in and near S gamma1, and levels of both were greatly increased in S mu(-/-) B cells. Finally, S mu(-/-) B cells underwent downstream CSR between S gamma1 and S epsilon on alleles that lacked S mu CSR to these sequences. Our findings show that AID targets downstream S regions independently of CSR with Smu and implicate an alternative pathway for IgE class switching that involves generation and joining of DSBs within two different downstream S regions before S mu joining.

  16. CD5-positive B-cell malignancies frequently express cross-reactive idiotypes associated with IgM autoantibodies.

    PubMed Central

    Kipps, T. J.; Robbins, B. A.; Tefferi, A.; Meisenholder, G.; Banks, P. M.; Carson, D. A.

    1990-01-01

    Using monoclonal antibodies (MAb) specific for cross-reactive idiotypes (CRIs) associated with human monoclonal IgM autoantibodies, we examined 57 biopsy specimens that previously had been noted to have immunohistologic features of CD5-positive B-cell small lymphocytic (SL) non-Hodgkin's lymphoma (NHL). Twenty-five lymphoma specimens were noted to be from patients with chronic lymphocytic leukemia (CLL). Eight of thirty-four (24%) immunoglobulin (Ig) kappa light-chain expressing lymphomas reacted with 17.109, a MAb specific for a major CRI encoded by a conserved Ig kappa variable region gene (Vk gene) of the VkIIIb sub-subgroup. All 17.109-reactive tissues and two 17.109-negative specimens were recognized by another MAb specific for VkIIIb framework determinant(s). Seven of all fifty-six (13%) Ig-expressing tumors bound G6, a MAb specific for an autoantibody heavy-chain-associated CRI that is encoded by a conserved antibody heavy chain variable region gene(s) (VHgene) of the VH1 subgroup. All seven G6-positive lymphomas and two G6-negative tumors reacted with Cc1, another MAb specific for a rheumatoid factor heavy-chain-associated CRI. A third autoantibody-heavy-chain-associated CRI, termed Lc1, was expressed by seven (13%) other lymphomas. Finally, a fourth MAb specific for RF heavy-chain-associated CRI, named B6, detected two additional tumors. The expression frequencies of autoantibody-associated CRIs among SL NHL patients without peripheral lymphocytosis did not differ from those noted among patients with CLL but were significantly higher than those observed among patients with NHL of follicular center-cell origin. These data imply that the malignant B cells of patients with either CD5-positive B-cell SL NHL or CLL express a restricted set of Ig V genes that have not substantially diversified from the germline DNA. Images Figure 1 PMID:1691593

  17. Circulating Natural IgM Antibodies Against Angiogenin in the Peripheral Blood Sera of Patients with Osteosarcoma as Candidate Biomarkers and Reporters of Tumorigenesis

    PubMed Central

    Savitskaya, Yulia A.; Rico, Genaro; Linares, Luis; González, Roberto; Téllez, René; Estrada, Eréndira; Marín, Norma; Martínez, Elisa; Alfaro, Alfonso; Ibarra, Clemente

    2010-01-01

    Background: Tumor immunology research has led to the identification of a number of tumor-associated self antigens, suggesting that most tumors trigger an immunogenic response, as is the case in osteosarcoma, where the detection of natural serum IgM antibodies might achieve the diagnosis of osteosarcoma. Natural IgM antibodies to tumor-associated proteins may expand the number of available tumor biomarkers for osteosarcoma and may be used together in a serum profile to enhance test sensitivity and specificity. Natural IgM antibodies can be consistently detected in the peripheral blood sera months to years before the tumor is diagnosed clinically. The study of the level of a potential biomarker many months (or years) prior to diagnosis is fundamentally important. Integrated circulating and imaging markers in clinical practice treating osteosarcoma have potential applications for controlling tumor angiogenesis. Objectives: To study the expression of natural IgM antibodies to the tumor antigens of angiogenesis in the peripheral blood sera of osteosarcoma patients and healthy individuals, and to develop serum-based predictive biomarkers. Methods: Peripheral venous blood samples were collected from 117 osteosarcoma patients and 117 patients with other tumors. All diagnosis was histologically confirmed. Staging of patients was performed according to the Enneking Surgical Staging System. The control group consisted of 117 age- and sex- matched healthy individuals. In this study, novel immunoconjugates were designed, synthesized and then used to develop a rapid, specific and sensitive enzyme-linked immunosorbent assay (ELISA) method to detect angiogenin (ANG)–IgM directly in the peripheral blood sera of humans. Results: Serum ANG–IgM levels are significantly higher in osteosarcoma patients than in healthy individuals (P < 0.005). Serum ANG–IgM levels varied widely, but were highly dependent on the concentration of IgM (r = 0.85; P < 0.0005). We found ANG–IgM in the

  18. VH and VL Domains of Polyspecific IgM and Monospecific IgG Antibodies Contribute Differentially to Antigen Recognition and Virus Neutralization Functions.

    PubMed

    Pasman, Y; Kaushik, A K

    2016-07-01

    We analysed contributions of variable heavy (FdVH ) and variable light (FdVL ) domains in comparison to scFv (FdVH +FdVL ) of naturally occurring polyspecific bovine IgM with an exceptionally long CDR3H and an induced monospecific bovine herpes virus-1 (BoHV-1) neutralizing IgG1 antibody in the context of to antigen-binding site and antibody function. Various recombinant FdVH , FdVL and scFv were constructed and expressed in Pichia pastoris from the bovine IgM and IgG1 antibody encoding cDNA. The scFv1H12 showed polyspecific antigen binding similar to parent IgM antibody, though subtle differences, for example, higher thyroglobulin recognition. Such differences reflect influence of the constant region on the antigen-binding site configuration. Unlike, variable light domain FdVL 1H12, the variable heavy domain FdVH 1H12 alone recognized multiple antigens that differed from the recognition pattern of scFv1H12 (FdVH +FdVL ) and the parent IgM antibody. Nonetheless, role of FdVL 1H12 in providing structural support to FdVH in antigen recognition is noted, apart from its intrinsic antigen recognition ability. Surface plasmon resonance analysis revealed low to moderate affinity of scFv1H12 to IgG antigen. By contrast, the individual FdVH 073 and FdVL 074, originating from induced BoHV-1 neutralizing IgG1 antibody, recognized target epitope on BoHV-1 weakly when compared to FdVH +FdVL (scFv3-18L). Interestingly, both the FdVH and FdVL domains of induced IgG antibody are required to achieve BoHV-1 neutralization. To conclude, there exist subtle functional differences in the contribution of FdVH and FdVL to antigen-binding site generation of polyspecific IgM and monospecific IgG antibodies relevant to antigen recognition and virus neutralization functions.

  19. Diagnosis of Leptospirosis: Comparison between Microscopic Agglutination Test, IgM-ELISA and IgM Rapid Immunochromatography Test

    PubMed Central

    Niloofa, Roshan; Fernando, Narmada; de Silva, Nipun Lakshitha; Karunanayake, Lilani; Wickramasinghe, Hasith; Dikmadugoda, Nandana; Premawansa, Gayani; Wickramasinghe, Rajitha; de Silva, H. Janaka; Premawansa, Sunil; Rajapakse, Senaka; Handunnetti, Shiroma

    2015-01-01

    Background Leptospirosis is diagnosed on clinical grounds, and confirmed by microscopic agglutination test (MAT). IgM-ELISA (Serion-Virion) and immunochromatography test (Leptocheck-WB) are two immunodiagnostic assays for leptospirosis. Their sensitivity, specificity and applicability in Sri Lanka have not been systematically evaluated. Methods Clinically diagnosed leptospirosis patients (n = 919) were recruited from three hospitals in the Western Province of Sri Lanka, during June 2012 to December 2013. MAT, IgM-ELISA and Leptocheck-WB were performed on all patient sera. MAT titer of ≥400 in single sample, four-fold rise or seroconversion ≥100 in paired samples were considered as positive for MAT. For diagnostic confirmation, MAT was performed during both acute and convalescent phases. Anti-leptospiral IgM ≥20 IU/ml and appearance of a band in the test window were considered as positive for IgM-ELISA and Leptocheck-WB test respectively. Patients with an alternative diagnosis (n = 31) were excluded. Data analysis was performed using two methods, i) considering MAT as reference standard and ii) using Bayesian latent class model analysis (BLCM) which considers each test as imperfect. Results MAT, IgM-ELISA and Leptocheck-WB positivity were 39.8%, 45.8% and 38.7% respectively during the acute phase. Acute-phase MAT had specificity and sensitivity of 95.7% and 55.3% respectively, when compared to overall MAT positivity. IgM-ELISA and Leptocheck-WB had similar diagnostic sensitivity when compared with acute-phase MAT as the gold standard, although IgM-ELISA showed higher specificity (84.5%) than Leptocheck-WB (73.3%). BLCM analysis showed that IgM-ELISA and Leptocheck-WB had similar sensitivities (86.0% and 87.4%), while acute-phase MAT had the lowest sensitivity (77.4%). However, acute-phase MAT had high specificity (97.6%), while IgM-ELISA and Leptocheck-WB showed similar but lower specificity (84.5% and 82.9%). Conclusions Both IgM-ELISA and Leptocheck-WB shows

  20. Characterization of RAG1 and IgM (mu chain) marking development of the immune system in red-spotted grouper (Epinephelus akaara).

    PubMed

    Mao, Ming-Guang; Lei, Ji-Lin; Alex, Perálvarez-Marín; Hong, Wan-Shu; Wang, Ke-Jian

    2012-10-01

    In vertebrates, lymphoid-specific recombinase protein encoded by recombination-activating genes (RAG1/2) plays a key role in V(D)J recombination of the T-cell receptor and B-cell receptor. In this study, both RAG1 and the immunoglobulin M (IgM) mu chain were cloned to characterize their potential role in the immune defense at developmental stages of red-spotted grouper, Epinephelus akaara. The open reading frame (ORF) of E. akaara RAG1 included 2778 nucleotide residues encoding a putative protein of 925 amino acids, while the ORF of the IgM mu chain had 1734 nucleotide residues encoding 578 amino acids including variable (VH) and constant (CH1-CH2-CH3-CH4) regions. E. akaara RAG1 was composed of a zinc-binding dimerization domain (ZDD) with a RING finger and zinc finger A (ZFA) in the non-core region and a nonamer-binding region (NBR), with a zinc finger B (ZFB), the central and C-terminal domains in the core region. Tridimensional models of the ZDD and NBR of E. akaara RAG1 were constructed for the first time in fishes, while a 3D model of the E. akaara IgM mu chain was also clarified. The RAG1 mRNA was only detected in the thymus and kidney of 4-month and 1.5-year old groupers using qPCR, and the RAG1 protein was confirmed using western blotting and immunohistochemistry. The IgM mu mRNA was examined in most tissues except the gonad. RAG1 and IgM mu gene expression were observed at 15 dph (days post-hatching) and 23 dph respectively, and increased to a higher level at 37 dph. In addition, this was the first time that the morphology of the E. akaara thymus was characterized. The oval-shaped thymus of 4-month old fish was clearly seen and there were amounts of T lymphocytes present. The results suggested that the immune action of E. akaara was likely to start to develop around 15 dph to 29 dph. The transcript level of the RAG1 gene and the number of lymphocytes in the thymus between 4-month and 1.5-year old groupers indicated that age-related thymic

  1. The amino-terminal domain of ORF149 of koi herpesvirus is preferentially targeted by IgM from carp populations surviving infection.

    PubMed

    Torrent, F; Villena, A; Lee, P A; Fuchs, W; Bergmann, S M; Coll, J M

    2016-10-01

    Recombinantly expressed fragments of the protein encoded by ORF149 (pORF149), a structural protein from the common- and koi-carp-infecting cyprinid herpesvirus-3 (CyHV-3) that was previously shown to be antigenic, were used to obtain evidence that its amino-terminal part contains immunodominant epitopes in fish populations that survived the infection. To obtain such evidence, nonspecific binding of carp serum tetrameric IgM had to be overcome by a novel ELISA protocol (rec2-ELISA). Rec2-ELISA involved pre-adsorption of carp sera with a heterologous recombinant fragment before incubation with pORF149 fragments and detection with anti-carp IgM monoclonal antibodies. Only in this way was it possible to distinguish between sera from uninfected and survivor carp populations. Although IgM from survivors recognised pORF149 fragments to a lesser degree than whole virus, specificity was confirmed by correlation of rec2- and CyHV-3-ELISAs, inhibition of rec2-ELISA by an excess of frgIIORF149, ELISA using IgM-capture, Western blotting, and reduction of reactivity in CyHV-3-ELISA by pre-adsorption of sera with frgIIORF149. The similarity of IgM-binding profiles between frgIORF149 (amino acid residues 42-629) and frgIIORF149 (42-159) and their reactivities with previously described anti-CyHV-3 monoclonal antibodies confirmed that most pORF149 epitopes were localised in its amino-terminal part. PMID:27383208

  2. Diagnostic Accuracy of the InBios Scrub Typhus Detect Enzyme-Linked Immunoassay for the Detection of IgM Antibodies in Northern Thailand

    PubMed Central

    Blacksell, Stuart D.; Tanganuchitcharnchai, Ampai; Nawtaisong, Pruksa; Kantipong, Pacharee; Laongnualpanich, Achara; Day, Nicholas P. J.

    2015-01-01

    In this study, we examined the diagnostic accuracy of the InBios Scrub Typhus Detect IgM enzyme-linked immunosorbent assay (ELISA) and determined the optimal diagnostic optical density (OD) cutoffs for screening and diagnostic applications based on prospectively collected, characterized samples from undifferentiated febrile illness patients in northern Thailand. Direct comparisons with the serological gold standard, indirect immunofluorescence assay (IFA), revealed strong statistical correlation of ELISA OD values and IFA IgM titers. Determination of the optimal ELISA cutoff for seroepidemiology or screening purposes compared to the corresponding IFA reciprocal titer of 400 as previously described for Thailand was 0.60 OD, which corresponded to a sensitivity (Sn) of 84% and a specificity (Sp) of 98%. The diagnostic performance against the improved and more-stringent scrub typhus infection criteria (STIC), correcting for low false-positive IFA titers, resulted in an Sn of 93% and an Sp of 91% at an ELISA cutoff of 0.5 OD. This diagnostic ELISA cutoff corresponds to IFA reciprocal titers of 1,600 to 3,200, which greatly reduces the false-positive rates associated with low-positive IFA titers. These data are in congruence with the recently improved serodiagnostic positivity criteria using the Bayesian latent class modeling approach. In summary, the InBios Scrub Typhus Detect IgM ELISA is affordable and easy-to-use, with adequate diagnostic accuracy for screening and diagnostic purposes, and should be considered an improved alternative to the gold standard IFA for acute diagnosis. For broader application, regional cutoff validation and antigenic composition for consistent diagnostic accuracy should be considered. PMID:26656118

  3. Single or combined effects of Lactobacillus sakei and inulin on growth, non-specific immunity and IgM expression in leopard grouper (Mycteroperca rosacea).

    PubMed

    Reyes-Becerril, Martha; Ascencio, Felipe; Gracia-Lopez, Vicente; Macias, Ma Esther; Roa, Marcos Cadena; Esteban, María Ángeles

    2014-08-01

    The aim of this study was to evaluate the single or combined effects of Lactobacillus sakei with inulin suitable for immunological in vivo studies in farmed fish. By in vitro assays, L. sakei strain 5-4 showed antibacterial activities against all assayed fish pathogens (except the Vibrio harveyi strain CAIM-1793). L. sakei was able to survive at high fish bile concentrations. Fermentation of the agave inulin resulted in a large increase in number of lactobacilli. For the in vivo study, fish were fed for 8 weeks four practical diets: control diet (control), L. sakei 5-4 (10(7) CFU/g), inulin (1% or 10 g/kg) and L. sakei + inulin (10(7) CFU/g + 10 g/kg). The weight gain showed clearly the synergistic effect of L. sakei 5-4 and inulin at 6 and 8 weeks of treatments. Leopard grouper fed with L. sakei alone or combined with inulin have significantly increased the assayed physiological and humoral immune parameters. By real-time PCR assays, the mRNA transcripts of immunoglobulin M (IgM) were found to be higher expressed in intestine, head kidney, mucus, gill, spleen and skin. Moreover, mRNA expression levels of IgM in head kidney and anterior intestine were measured by real-time PCR. L. sakei 5-4 and L. sakei + inulin supplemented diet up-regulated the expression of IgM at week 4 and 8 in intestine and head kidney, respectively. These results support the idea that the L. sakei 5-4 alone or combined with agave inulin improved growth performance and stimulates the immune system of leopard grouper.

  4. Expansion of natural killer cells in mice transgenic for IgM antibody to ganglioside GD2: demonstration of prolonged survival after challenge with syngeneic tumor cells.

    PubMed

    Kawashima, Ikuo; Yoshida, Yukiko; Taya, Chouji; Shitara, Hiroshi; Yonekawa, Hiromichi; Karasuyama, Hajime; Tada, Nobuhiko; Furukawa, Koichi; Tai, Tadashi

    2003-08-01

    IgM antibodies to gangliosides, sialic acid-containing glycosphingolipids, have been shown to mediate anti-tumor effects in cancer patients with melanoma and neuroblastoma and to correlate with survival. Mechanisms by which the antibodies induce tumor suppression, however, have not been systematically studied. To investigate this point, we produced and characterized C57BL/6 mice transgenic for IgM antibody to ganglioside GD2. The transgenic (TG) mice showed high IgM, but not IgG antibody titers against GD2 in their sera. No significant clinical symptoms were observed. When EL4 cells, syngeneic T lymphoma that express ganglioside GD2, were injected into TG mice, prolonged survival was observed. Complement-dependent cytotoxicity (CDC) of EL4 cells was mediated with TG mice sera. Neither antibody-dependent cellular cytotoxicity with their sera nor cytotoxic T lymphocyte activity to EL4 cells was shown in TG mice. Spleen lymphocytes from TG mice had increased numbers of natural killer (NK) cells, but not T cells, B cells, or macrophages compared with wild-type mice. Depletion of NK cells with anti-asialo GM1 rabbit serum reduced or abrogated the observed anti-tumor effects, suggesting that NK cells play a major role in tumor eradication or suppression. NK cell activity in TG mice was much higher than wild-type mice. Moreover, TG mice showed prolonged survival after injection with syngeneic B16 melanoma cells, which express GM3, but not GD2 or GD3. Taking these results together, our studies demonstrate that the TG mice have significant anti-tumor characteristics, probably due to CDC and NK cell expansion and activation with anti-ganglioside GD2 antibody.

  5. Detection of IgE, IgG, IgA and IgM antibodies against raw and processed food antigens

    PubMed Central

    Vojdani, Aristo

    2009-01-01

    Background Despite the first documented case of food allergy to cooked food in 1921 by Prausnitz and Kustner, all commercial food antigens are prepared from raw food. Furthermore, all IgE and IgG antibodies against dietary proteins offered by many clinical laboratories are measured against raw food antigens. Methods We developed an enzyme-linked immunosorbent assay for the measurement of IgE, IgG, IgA and IgM antibodies against raw and processed food antigens. Sera with low or high reactivity to modified food antigens were subjected to myelin basic protein, oxidized low density lipoprotein, and advanced glycation end products (AGE) such as AGE-human serum albumin and AGE-hemoglobin. Results Compared to raw food antigens, IgE antibodies showed a 3–8-fold increase against processed food antigens in 31% of the patients. Similarly, IgG, IgA and IgM antibodies against modified food antigens overall were found at much higher levels than antibody reactions against raw food antigens. Almost every tested serum with high levels of antibodies against modified food antigens showed very high levels of antibodies against myelin basic protein, oxidized low density lipoprotein, AGE-human serum albumin and AGE-hemoglobin. Conclusion We conclude that the determination of food allergy, intolerance and sensitivity would be improved by testing IgE, IgG, IgA and IgM antibodies against both raw and processed food antigens. Antibodies against modified food antigens, by reacting with AGEs and tissue proteins, may cause perturbation in degenerative and autoimmune diseases such as diabetes, atherosclerosis, inflammation, autoimmunity, neurodegeneration and neuroautoimmunity. PMID:19435515

  6. The amino-terminal domain of ORF149 of koi herpesvirus is preferentially targeted by IgM from carp populations surviving infection.

    PubMed

    Torrent, F; Villena, A; Lee, P A; Fuchs, W; Bergmann, S M; Coll, J M

    2016-10-01

    Recombinantly expressed fragments of the protein encoded by ORF149 (pORF149), a structural protein from the common- and koi-carp-infecting cyprinid herpesvirus-3 (CyHV-3) that was previously shown to be antigenic, were used to obtain evidence that its amino-terminal part contains immunodominant epitopes in fish populations that survived the infection. To obtain such evidence, nonspecific binding of carp serum tetrameric IgM had to be overcome by a novel ELISA protocol (rec2-ELISA). Rec2-ELISA involved pre-adsorption of carp sera with a heterologous recombinant fragment before incubation with pORF149 fragments and detection with anti-carp IgM monoclonal antibodies. Only in this way was it possible to distinguish between sera from uninfected and survivor carp populations. Although IgM from survivors recognised pORF149 fragments to a lesser degree than whole virus, specificity was confirmed by correlation of rec2- and CyHV-3-ELISAs, inhibition of rec2-ELISA by an excess of frgIIORF149, ELISA using IgM-capture, Western blotting, and reduction of reactivity in CyHV-3-ELISA by pre-adsorption of sera with frgIIORF149. The similarity of IgM-binding profiles between frgIORF149 (amino acid residues 42-629) and frgIIORF149 (42-159) and their reactivities with previously described anti-CyHV-3 monoclonal antibodies confirmed that most pORF149 epitopes were localised in its amino-terminal part.

  7. Single or combined effects of Lactobacillus sakei and inulin on growth, non-specific immunity and IgM expression in leopard grouper (Mycteroperca rosacea).

    PubMed

    Reyes-Becerril, Martha; Ascencio, Felipe; Gracia-Lopez, Vicente; Macias, Ma Esther; Roa, Marcos Cadena; Esteban, María Ángeles

    2014-08-01

    The aim of this study was to evaluate the single or combined effects of Lactobacillus sakei with inulin suitable for immunological in vivo studies in farmed fish. By in vitro assays, L. sakei strain 5-4 showed antibacterial activities against all assayed fish pathogens (except the Vibrio harveyi strain CAIM-1793). L. sakei was able to survive at high fish bile concentrations. Fermentation of the agave inulin resulted in a large increase in number of lactobacilli. For the in vivo study, fish were fed for 8 weeks four practical diets: control diet (control), L. sakei 5-4 (10(7) CFU/g), inulin (1% or 10 g/kg) and L. sakei + inulin (10(7) CFU/g + 10 g/kg). The weight gain showed clearly the synergistic effect of L. sakei 5-4 and inulin at 6 and 8 weeks of treatments. Leopard grouper fed with L. sakei alone or combined with inulin have significantly increased the assayed physiological and humoral immune parameters. By real-time PCR assays, the mRNA transcripts of immunoglobulin M (IgM) were found to be higher expressed in intestine, head kidney, mucus, gill, spleen and skin. Moreover, mRNA expression levels of IgM in head kidney and anterior intestine were measured by real-time PCR. L. sakei 5-4 and L. sakei + inulin supplemented diet up-regulated the expression of IgM at week 4 and 8 in intestine and head kidney, respectively. These results support the idea that the L. sakei 5-4 alone or combined with agave inulin improved growth performance and stimulates the immune system of leopard grouper. PMID:24464476

  8. IgM, IgG and IgA rheumatoid factors and circulating immune complexes in patients with AIDS and AIDS-related complex with serological abnormalities.

    PubMed Central

    Procaccia, S; Lazzarin, A; Colucci, A; Gasparini, A; Forcellini, P; Lanzanova, D; Foppa, C U; Novati, R; Zanussi, C

    1987-01-01

    To investigate some humoral aspects which may reflect the involvement of B lymphocytes in the acquired immunodeficiency syndrome (AIDS), we used an enzyme-linked immunoassay (ELISA) to determine the levels of IgM, IgG and IgA rheumatoid factors (RF) in 16 patients suffering from full-blown AIDS and 32 patients with AIDS-related complex (ARC), in the clinical form of lymphoadenopathy syndrome (LAS), compared with 40 healthy, young heterosexual subjects. Both AIDS and ARC patients showed a greater incidence of high IgM RF levels, with mean values significantly higher than controls, but with no differences between the two pathological groups. IgG RF behaviour was similar in the two patient populations and the healthy subjects. IgA RF were significantly raised in AIDS and ARC. Further information on RF was obtained by determination of the immunoglobulin levels of the respective isotypes in the same patients. Mean IgG levels were above normal in AIDS and ARC patients, but the latter group showed a higher incidence of increased values and higher mean levels. The IgA isotype was significantly increased mainly in AIDS patients. The behaviour of IgM was virtually the same in the three groups studied. A difference between AIDS and ARC patients was established by the detection of circulating immune-complexes (IC) by the C1q-binding and CIC-conglutinin assays. IC were significantly high, by both methods, only in the ARC group, but normal or very low in AIDS. These overall findings suggest once again the impairment of B cell function in AIDS, with prevalent hyperactivation in ARC and exhaustion in full-blown AIDS, and apparent preservation, in the latter group, of the antibody responses which are more closely related to the activity of subsets of T helper cells. PMID:3608224

  9. Identification of IgG alloantibodies in patients with high-titer IgM cold agglutinins by serum/plasma affinity chromatography.

    PubMed

    Stahl, D; Kreft, H; Hack, H; Schraven, B; Roelcke, D

    1997-01-01

    The detection of IgG alloantibodies in the presence of high-titer cold autoagglutinins (CAs) can be extremely difficult, especially under pressure of time when transfusion of red blood cells is urgently needed. Here we demonstrate that IgG alloantibodies in the presence of high-titer IgM CAs can be easily detected by quantitative IgG purification from serum or plasma by affinity chromatography. In comparison with the routinely used methods for IgG alloantibody identification, affinity chromatography shows better or identical results and is the method leading to results most rapidly.

  10. Therapeutic Strategies for Human IgM Antibodies Directed at Tumor-Associated Ganglioside Antigens: Discoveries Made During the Morton Era and Future Directions.

    PubMed

    Jones, Peter C; Irie, Reiko F

    2016-01-01

    Tumor-associated gangliosides have been investigated for their potential as antigenic targets for more than 35 years, culminating in the recent Food and Drug Administration approval of dinutuximab (Unituxin), an IgG antibody targeted against GD2, for the treatment of neuroblastoma in children. This review is focused on discoveries and development of therapeutic approaches involving human IgM antibodies directed against gangliosides, which occurred over the past 40 years at University of California-Los Angeles and the John Wayne Cancer Institute, where Dr. Donald Morton led the surgical oncology department until his death. PMID:27481004

  11. Serological investigation of the prevalence of anti-dengue IgM and IgG antibodies in Attapeu Province, South Laos.

    PubMed

    Peyerl-Hoffmann, G; Schwöbel, B; Jordan, S; Vamisaveth, V; Phetsouvanh, R; Christophel, E M; Phompida, S; Sonnenburg, F V; Jelinek, T

    2004-02-01

    The prevalence of dengue antibodies was determined in the Attapeu region of South Laos with 225 blood samples collected from mostly febrile patients during the rainy season August - October 2001. An IgM capture ELISA was positive for one (0.4%) sample, while 177 (79%) samples were positive in an indirect IgG ELISA. Of the positive IgG samples, 20 (11.3%) were also positive on blood slides for Plasmodium falciparum. Dengue fever seems to be widespread in this area, but clinical dengue diagnosis remains difficult, especially in the first days of illness when physicians have to discriminate between dengue and other febrile illnesses.

  12. Variable region sequences of murine IgM anti-IgG monoclonal autoantibodies (rheumatoid factors). II. Comparison of hybridomas derived by lipopolysaccharide stimulation and secondary protein immunization

    PubMed Central

    1987-01-01

    We have obtained the complete variable region mRNA sequences of 11 LPS- derived and 14 secondary immunization-derived monoclonal IgM anti-IgG antibodies (rheumatoid factors, RFs). A comparative analysis of these sequences showed that monoclonal RFs derived after polyclonal activation are structurally very similar to RFs derived after secondary protein immunization. This study was undertaken to evaluate the potential relationship between two previously described phenomena: (a) during a secondary response to a protein antigen, RF is produced in quantities that equal or exceed the immunogen-specific antibody; and (b) the frequency of B cells that make RF after polyclonal activation is quite high; 3-10%. It has been unclear whether LPS-stimulated cells that produce IgM anti-IgG that is detected by an in vitro assay are related to the cells that produce RF after in vivo stimulation. The similarity of the antigen receptors found in the two types of RF, however, suggests that most or all of the RF-producing B cells detected after LPS stimulation would also be stimulated during the secondary immune response. Thus, the presence of relatively large number of B cells that can make RF after nonspecific stimulation provides an explanation for the magnitude of RF production accompanying the secondary immune response. PMID:3494096

  13. Terminal carbohydrate composition, IgM level and enzymatic and bacteriostatic activity of European sea bass (Dicentrarchus labrax) skin epidermis extracts.

    PubMed

    Guardiola, Francisco A; de Haro, Juan P; Díaz-Baños, Francisco Guillermo; Meseguer, José; Cuesta, Alberto; Esteban, M Ángeles

    2015-11-01

    Although the skin is one of the main defense barriers of fish to date, very little is known about the immune implications and the properties of the numerous substances present in skin cells. In the present study, terminal carbohydrate composition and some components of the skin immunity (total IgM level, and several enzymatic and bacteriostatic activities) present on aqueous and organic epidermal extracts of European sea bass (Dicentrarchus labrax) were determined. Most of the parameters measured followed a protein concentration dose-response. Curiously, both skin extracts have similar levels of total IgM. However, aqueous extracts showed higher presence of some terminal carbohydrates, alkaline phosphatase and esterase activities and lower proteases and ceruloplasmin activities than epidermal organic extracts. Regarding the bacteriostatic activity, the growth of all the bacterial strains tested was reduced when cultivated in presence of organic extracts, being the observed reduction correlated to the protein concentration present in the extract sample. On the contrary, skin aqueous extracts have no significant effect on bacterial growth or even allow bacteria to overgrow, suggesting that the bacteria could use the extracts as a nutrient source. The results are discussed and compared with the same activities studied on fish skin mucus in order to understand their possible implications on mucosal immunity.

  14. In Men at Risk of HIV Infection, IgM, IgG1, IgG3 and IgA Reach the Human Foreskin Epidermis

    PubMed Central

    Lemos, Maria P.; Karuna, Shelly T.; Mize, Gregory J.; Fong, Youyi; Montano, Silvia M.; Ganoza, Carmela; Lama, Javier R.; Sanchez, Jorge; McElrath, M. Juliana

    2015-01-01

    We profiled the humoral response in the penis, an area that has been minimally explored but may be relevant for protecting insertive men against HIV and other sexually-acquired infections. Comparing paired tissue samples from 20 men at risk of HIV infection, foreskin contains less IgA and more IgG2 than colon. Using foreskin dermal and epidermal explants and paired plasma from 17 men, we examined Ig accumulation by normalizing Ig to human serum albumin (HSA) transudation. Dermal IgM, IgG2, IgA, and IgE ratios were greater than in plasma, suggesting there is local antibody secretion at the dermis. Local Ig transcription was concentrated at the inner rather than the outer foreskin, and inner foreskin Ig ratios did not correlate with blood, indicating that localized production can contribute to the foreskin response. IgM, IgG1, IgG3, and IgA have preferential access to the foreskin epidermis, whereas IgG2, IgG4, and IgE are restricted to the dermis. Lastly, Ad5-specific IgA was selectively in the colon; whereas foreskin Ad5 IgG was mainly derived from blood, and reached the inner epidermis at higher ratios than the outer (p<0.002). In summary, the foreskin antibody response combines local and systemic sources and there is selective isotype accumulation in the epidermis. PMID:26509877

  15. High levels of IgM antibodies specific for Toxoplasma gondii in pregnancy 12 years after primary toxoplasma infection. Case report.

    PubMed

    Bobić, B; Sibalić, D; Djurković-Djaković, O

    1991-01-01

    A case of high levels of specific IgM antibodies registered by the immunosorbent agglutination assay (ISAgA) in a pregnant woman with a history of toxoplasmosis is reported. The patient had acute lymphoglandular toxoplasmosis diagnosed serologically by increases in the specific antibody titer detected by the Sabin-Feldman test (SFT) and pathohistologically 12 years before pregnancy. In pregnancy, she had stable titers of specific antibodies registered by the SFT, enzyme-linked immunosorbent assay (ELISA), indirect fluorescent antibody test (IFAT), indirect hemagglutination test (IHAT) and direct agglutination test. Specific IgM antibodies were detected by the ISAgA but not by the IgM-IFAT, IgM-ELISA and IgM-IHAT. She had a normal pregnancy and gave birth to a clinically healthy baby who had a negative ISAgA finding at 7 days of age. This case indicates that ISAgA is not necessarily a marker of recent infection and is therefore not reliable for the diagnosis of toxoplasmosis in pregnancy.

  16. In men at risk of HIV infection, IgM, IgG1, IgG3, and IgA reach the human foreskin epidermis.

    PubMed

    Lemos, M P; Karuna, S T; Mize, G J; Fong, Y; Montano, S M; Ganoza, C; Lama, J R; Sanchez, J; McElrath, M J

    2016-05-01

    We profiled the humoral response in the penis, an area that has been minimally explored but may be relevant for protecting insertive men against HIV and other sexually acquired infections. Comparing paired tissue samples from 20 men at risk of HIV infection, foreskin contains less immunoglobulin A (IgA) and more IgG2 than colon. Using foreskin dermal and epidermal explants and paired plasma from 17 men, we examined Ig accumulation by normalizing Ig to human serum albumin (HSA) transudation. Dermal IgM, IgG2, IgA, and IgE ratios were greater than that in plasma, suggesting there is local antibody secretion at the dermis. Local Ig transcription was concentrated at the inner rather than the outer foreskin, and inner foreskin Ig ratios did not correlate with blood, indicating that localized production can contribute to the foreskin response. IgM, IgG1, IgG3, and IgA have preferential access to the foreskin epidermis, whereas IgG2, IgG4, and IgE are restricted to the dermis. Lastly, Ad5-specific IgA was selectively present in the colon, whereas foreskin Ad5 IgG was mainly derived from blood, and reached the inner epidermis at higher ratios than the outer (P<0.002). In summary, the foreskin antibody response combines local and systemic sources, and there is selective isotype accumulation in the epidermis. PMID:26509877

  17. Development of novel immunoglobulin G (IgG), IgA, and IgM enzyme immunoassays based on recombinant Puumala and Dobrava hantavirus nucleocapsid proteins.

    PubMed

    Meisel, Helga; Wolbert, Anne; Razanskiene, Ausra; Marg, Andreas; Kazaks, Andris; Sasnauskas, Kestutis; Pauli, Georg; Ulrich, Rainer; Krüger, Detlev H

    2006-12-01

    Human infections with Asian and European hantaviruses can result in hemorrhagic fever with renal syndromes of differing severities characterized by renal dysfunction and sometimes by pulmonary symptoms. For the serological detection of human infections by hantaviruses relevant for Europe, we developed monoclonal antibody capture immunoglobulin G (IgG) and IgA enzyme-linked immunosorbent assays (ELISAs) based on yeast-expressed nucleocapsid proteins of Puumala and Dobrava hantaviruses. Moreover, for diagnosis of acute infections, mu-capture IgM ELISAs were established with nucleocapsid proteins expressed in Drosophila melanogaster Schneider S2 cells. The cutoff values of the ELISAs were determined by investigation of up to 500 human anti-hantavirus-negative serum samples. The specificities of the Puumala and Dobrava virus-specific IgM, IgA, and IgG ELISAs were found to be 100%. The sensitivities of these ELISAs were determined to be 100% with panels of characterized anti-Puumala or anti-Dobrava virus-positive human serum samples. In most cases, Puumala and Dobrava virus infections could be differentiated by ELISA reactivity alone, i.e., endpoint titration with homologous and heterologous antigens.

  18. The use of IgM tests for analysis of the causes of measles vaccine failures: experience gained in an epidemic in Hungary in 1980 and 1981.

    PubMed

    Nagy, G; Kósa, S; Takátsy, S; Koller, M

    1984-01-01

    Following a period of 6 years of low measles incidence, an epidemic occurred in Hungary with more than 11,000 reported cases between September 1980 and August 1981. About 60% of the cases had a documented history of previous measles vaccination. Serum samples obtained from 7815 patients were examined for measles antibody by haemagglutination inhibition (HI). In addition to conventional antibody titration, most of the sera or their IgM fraction obtained by a simple ion exchange chromatography were tested for the presence of measles-specific IgM antibodies by 2-mercaptoethanol (2-ME) treatment, and in 300 patients also by the fluorescent antibody (FA) technique. Laboratory results confirmed the diagnosis of measles in 5356 patients and supported it in 685 cases. Primary antibody response was found in 96.1% of unvaccinated and in 77.4% of previously vaccinated patients. The percentage of secondary antibody responses increased with increasing time from vaccination only in patients vaccinated before their first birthday, whereas in those who were immunized when over 12 months old, the distribution of primary and secondary antibody responses was independent from the time that had elapsed since vaccination. Therefore, secondary vaccine failure due to waning immunity account for only 6.2% of previously vaccinated patients, whereas in 93.8% of patients, including the majority of those with secondary antibody response, a primary failure of vaccination due to unsuccessful immunization was incriminated.

  19. Effects of (Anti) Androgenic Endocrine Disruptors (DEHP and Butachlor) on Immunoglobulin M (IgM) and Leukocytes Counts of Male Rainbow Trout (Oncorhynchus mykiss).

    PubMed

    Ahmadivand, Sohrab; Farahmand, Hamid; Mirvaghefi, Alireza; Eagderi, Soheil; Zargar, Ashkan

    2015-06-01

    The effect of two anti-androgenic endocrine disrupting compounds, i.e. the plasticizer di (2-ethylhexyl) phthalate (DEHP) and herbicide butachlor, were evaluated for their effects on immunoglobulin M (IgM) and leukocytes in male rainbow trout. Also, plasma testosterone (T) concentration was measured to confirm their anti-androgenic effects. In the first experiment, trout were treated with 50 mg/kg (body weight) DEHP intraperitoneally, and in the second one, fish were exposed to 0.39 mg/L butachlor for 10 days. The results showed that T concentrations and white blood cells were significantly lower in fish exposed to either DEHP or butachlor compared to control fish (p < 0.05). Fish showed significantly elevated neutrophil levels and decreased lymphocyte levels in the butachlor (p < 0.05); however, no significant difference was observed in lymphocyte and neutrophils values in the DEHP treatment (p > 0.05). In addition, no significant differences were found in IgM, eosinophil and monocyte parameters in either DEHP or butachlor treatments (p > 0.05). These results confirmed that leukocytes counts can be considered as a novel marker of immunotoxicity triggered by (anti) androgenic endocrine disruptors.

  20. Effects of (Anti) Androgenic Endocrine Disruptors (DEHP and Butachlor) on Immunoglobulin M (IgM) and Leukocytes Counts of Male Rainbow Trout (Oncorhynchus mykiss).

    PubMed

    Ahmadivand, Sohrab; Farahmand, Hamid; Mirvaghefi, Alireza; Eagderi, Soheil; Zargar, Ashkan

    2015-06-01

    The effect of two anti-androgenic endocrine disrupting compounds, i.e. the plasticizer di (2-ethylhexyl) phthalate (DEHP) and herbicide butachlor, were evaluated for their effects on immunoglobulin M (IgM) and leukocytes in male rainbow trout. Also, plasma testosterone (T) concentration was measured to confirm their anti-androgenic effects. In the first experiment, trout were treated with 50 mg/kg (body weight) DEHP intraperitoneally, and in the second one, fish were exposed to 0.39 mg/L butachlor for 10 days. The results showed that T concentrations and white blood cells were significantly lower in fish exposed to either DEHP or butachlor compared to control fish (p < 0.05). Fish showed significantly elevated neutrophil levels and decreased lymphocyte levels in the butachlor (p < 0.05); however, no significant difference was observed in lymphocyte and neutrophils values in the DEHP treatment (p > 0.05). In addition, no significant differences were found in IgM, eosinophil and monocyte parameters in either DEHP or butachlor treatments (p > 0.05). These results confirmed that leukocytes counts can be considered as a novel marker of immunotoxicity triggered by (anti) androgenic endocrine disruptors. PMID:25708297

  1. Characterization of the gene for the membrane and secretory form of the IgM heavy-chain constant region gene (C mu) of the cow (Bos taurus).

    PubMed Central

    Mousavi, M; Rabbani, H; Pilström, L; Hammarström, L

    1998-01-01

    Our present understanding of the evolution of immunoglobulins is derived from a few vertebrate species. In order to obtain additional information on the development of the humoral immune system, we cloned and determined the nucleotide sequence of the bovine cDNA and genomic IgM heavy-chain constant region gene (C mu). The gene contains four constant region domain-encoding exons (CH1 to CH4) and two exons encoding the transmembrane domain (TM1, TM2), expressed in the membrane-bound receptor form of the IgM. The sequence of a cDNA clone encoding the 3' portion of the membrane form of the mu-chain revealed that the TM1 exon is spliced to the CH4 exon, as occurs in other mammals. Comparison of deduced amino acid sequence data from different vertebrates revealed a high similarity to sheep C mu (88%) and a lower degree of similarity to pig (62%), rat (62%), rabbit (58%) human (56%), hamster (55%), mouse (54%), chicken (28%) and horned shark (22%) C mu. Images Figure 6 Figure 7 PMID:9659232

  2. [Preliminary results of the use of an ELISA test for the differential determination of IgG and specific IgM in toxoplasmosis serology].

    PubMed

    Miceli, M D; Schifaudo, G; Vitale, G; Di Rosa, S; Mansueto, S

    1982-09-01

    The diagnosis of acute toxoplasmosis usually depends on serology, since clinical and/or histological features are difficult and/or often misleading. IgM titers are the best indicators of infection acquired in the past two to four months. IgG titers are generally correlated with non-active infections as well as previous (symptomatic and/or asymptomatic) illness. In the present paper, results obtained with a new kit for the evaluation of IgG and specific-IgM by micro-ELISA technique, are reported. Optical density values in sera from: a) blood donors; b) miscellany group; c) suspected toxoplasmosis, were at various degree high for IgG. Few sera of the entire sample (in the group of suspected toxoplasmosis and one in miscellany) showed high optical density for specific IgM. Reading of results with spectrophotometer were in agreement with reading to the naked eye. Reproducibility was satisfactory. Unfortunately some sera positive in the first determinations gave equivocal results in successive proofs. For these reasons diagnosis of toxoplasmosis must be based upon the use of at least three tests.

  3. Molecular components of the B cell antigen receptor complex of class IgD differ partly from those of IgM.

    PubMed Central

    Wienands, J; Hombach, J; Radbruch, A; Riesterer, C; Reth, M

    1990-01-01

    Two classes of immunoglobulin, IgM and IgD, are present as antigen receptors on the surface of mature B lymphocytes. We show here that IgD molecules are noncovalently associated in the B cell membrane with a heterodimer consisting of two proteins of 35 kd (IgD-alpha) and 39 kd (Ig-beta), respectively. The two novel proteins are not found in the IgD-expressing myeloma J558L delta m, which fails to bring IgD antigen receptor onto the cell surface. In a surface IgD positive variant line of this myeloma, however, membrane-bound IgD molecules are associated with the heterodimer, suggesting that the formation of an antigen receptor complex is required for surface IgD expression. We further demonstrate that the IgD-associated heterodimer differs partly from that of the IgM antigen receptor and that its binding to the heavy chain only requires the presence of the last constant domain and the transmembrane part of the delta m chain. Images Fig. 3. Fig. 5. Fig. 6. Fig. 7. PMID:2303036

  4. Four-layer enzyme immunoassay (EIA) detection of differences in IgG, IgM and IgA antibody response to Aujeszky's disease virus in infected and vaccinated pigs.

    PubMed

    Rodák, L; Smíd, B; Valícek, L; Jurák, E

    1987-02-01

    The use of the four-layer enzyme immunoassay (EIA) for the detection of IgG, IgM and IgA antibodies against Aujeszky's disease virus in blood and oropharyngeal swabs of infected and vaccinated pigs is described. Mean antibody titres obtained using the four-layer EIA were 6.1 and 3829 times higher compared with the indirect enzyme-linked immunosorbent assay (ELISA) and virus neutralization (VN) test, respectively. The VN test detected mainly IgG antibodies, while the IgM antibodies did not react. Using the EIA, the first antiviral antibodies in sera were demonstrated on Days 5-7 after infection or vaccination. Up to the 7th day, demonstrable antibodies were almost exclusively of the IgM class. In infected pigs high titres of IgM antibodies were still detected on Day 18, while in vaccinated animals they were absent by this time. Antibodies of the IgG class appeared in infected pigs sooner (Day 7) than in vaccinated pigs (Day 10) and reached higher mean titres. Antibodies of the IgA class were demonstrable from Day 10 only in samples from infected pigs. Similar antibody dynamics and distribution were detected in oropharyngeal swabs, except that the IgG and IgM titres were roughly 100 times lower than in sera. However, titres of IgA antibodies in oropharyngeal swabs were two times higher than in sera. The greatest differences between both groups of animals were recorded on Day 18; in the infected pigs, IgG, IgM and IgA antibodies were present in sera and oropharyngeal swabs at that time, while in vaccinated pigs only IgG antibodies were demonstrable. The effect of infection and vaccination on the pattern of the immune response as well as the importance of the detection of individual immunoglobulin classes for the specificity of the enzyme immunoassay are discussed.

  5. Spontaneous Development of IgM Anti-Cocaine Antibodies in Habitual Cocaine Users: Effect on IgG Antibody Responses to a Cocaine Cholera Toxin B Conjugate Vaccine

    PubMed Central

    Orson, Frank M.; Rossen, Roger D.; Shen, Xiaoyun; Lopez, Angel Y.; Wu, Yan; Kosten, Thomas R.

    2014-01-01

    Background and Objectives In cocaine vaccine studies, only a minority of subjects made strong antibody responses. To investigate this issue, IgG and IgM antibody responses to cocaine and to cholera toxin B (CTB—the carrier protein used to enhance immune responses to cocaine) were measured in sera from the 55 actively vaccinated subjects in a Phase IIb randomized double-blind placebo-controlled trial (TA-CD 109). Methods Isotype specific ELISAs were used to measure IgG and IgM anti-cocaine and anti-CTB antibody in serial samples collected prior to and at intervals after immunization. We assessed IgG anti-cocaine responses of patients with pre-vaccination IgM anti-cocaine antibodies. Competitive inhibition ELISA was used to evaluate antibody specificity. Results and Conclusions Before immunization, 36/55 subjects had detectable IgM antibodies to cocaine, and 9 had IgM levels above the 95% confidence limit of 11 µg/ml. These nine had significantly reduced peak IgG anti-cocaine responses at 16 weeks, and all were below the concentration (40 µg/ml) considered necessary to discourage recreational cocaine use. The IgG anti-CTB responses of these same subjects were also reduced. Scientific Significance Subjects who develop an IgM antibody response to cocaine in the course of repeated recreational exposure to this drug are significantly less likely to produce high levels of IgG antibodies from the cocaine conjugate vaccine. The failure may be due to recreational cocaine exposure induction of a type 2 T-cell independent immune response. Such individuals will require improved vaccines and are poor candidates for the currently available vaccine. PMID:23414504

  6. Monitoring of Anti-Hepatitis E Virus Antibody Seroconversion in Asymptomatically Infected Blood Donors: Systematic Comparison of Nine Commercial Anti-HEV IgM and IgG Assays.

    PubMed

    Vollmer, Tanja; Diekmann, Juergen; Eberhardt, Matthias; Knabbe, Cornelius; Dreier, Jens

    2016-08-22

    Diagnosis of hepatitis E virus (HEV) is usually determined serologically by detection of the presence of immunoglobulin (Ig)M antibodies or rising anti-HEV IgG titers. However, serological assays have demonstrated a significant variation in their sensitivities and specificities. In this study, we present the systematic comparison of different immunological anti-HEV assays using complete seroconversion panels of 10 virologically confirmed HEV genotype 3 infected individuals. Assay sensitivities were further evaluated by testing serially diluted World Health Organization (WHO) reference reagent for hepatitis E virus antibody and one patient sample infected with HEV genotype 3. Anti-HEV IgM and IgG antibody presence was determined using the immunological assays Wantai HEV IgM/IgG enzyme-linked immunosorbent assay (ELISA) (Sanbio, Uden, The Netherlands), recomWell HEV IgM/IgG (Mikrogen, Neuried, Germany), HEV IgM ELISA 3.0, HEV ELISA, HEV ELISA 4.0, Assure HEV IgM Rapid Test (all MP Biomedicals Europe, Illkirch Cedex, France) and Anti-HEV ELISA (IgM/IgG, Euroimmun, Lübeck, Germany). The assays showed differences regarding their analytical and diagnostic sensitivities, with anti-HEV IgM assays (n = 5) being more divergent compared to anti-HEV IgG (n = 4) assays in this study. Considerable variations were observed particularly for the detection period of IgM antibodies. This is the first study systematically characterizing serologic assays on the basis of seroconversion panels, providing sample conformity for a conclusive comparison. Future studies should include the assay comparison covering the four different genotypes.

  7. Monitoring of Anti-Hepatitis E Virus Antibody Seroconversion in Asymptomatically Infected Blood Donors: Systematic Comparison of Nine Commercial Anti-HEV IgM and IgG Assays

    PubMed Central

    Vollmer, Tanja; Diekmann, Juergen; Eberhardt, Matthias; Knabbe, Cornelius; Dreier, Jens

    2016-01-01

    Diagnosis of hepatitis E virus (HEV) is usually determined serologically by detection of the presence of immunoglobulin (Ig)M antibodies or rising anti-HEV IgG titers. However, serological assays have demonstrated a significant variation in their sensitivities and specificities. In this study, we present the systematic comparison of different immunological anti-HEV assays using complete seroconversion panels of 10 virologically confirmed HEV genotype 3 infected individuals. Assay sensitivities were further evaluated by testing serially diluted World Health Organization (WHO) reference reagent for hepatitis E virus antibody and one patient sample infected with HEV genotype 3. Anti-HEV IgM and IgG antibody presence was determined using the immunological assays Wantai HEV IgM/IgG enzyme-linked immunosorbent assay (ELISA) (Sanbio, Uden, The Netherlands), recomWell HEV IgM/IgG (Mikrogen, Neuried, Germany), HEV IgM ELISA 3.0, HEV ELISA, HEV ELISA 4.0, Assure HEV IgM Rapid Test (all MP Biomedicals Europe, Illkirch Cedex, France) and Anti-HEV ELISA (IgM/IgG, Euroimmun, Lübeck, Germany). The assays showed differences regarding their analytical and diagnostic sensitivities, with anti-HEV IgM assays (n = 5) being more divergent compared to anti-HEV IgG (n = 4) assays in this study. Considerable variations were observed particularly for the detection period of IgM antibodies. This is the first study systematically characterizing serologic assays on the basis of seroconversion panels, providing sample conformity for a conclusive comparison. Future studies should include the assay comparison covering the four different genotypes. PMID:27556482

  8. Monitoring of Anti-Hepatitis E Virus Antibody Seroconversion in Asymptomatically Infected Blood Donors: Systematic Comparison of Nine Commercial Anti-HEV IgM and IgG Assays.

    PubMed

    Vollmer, Tanja; Diekmann, Juergen; Eberhardt, Matthias; Knabbe, Cornelius; Dreier, Jens

    2016-01-01

    Diagnosis of hepatitis E virus (HEV) is usually determined serologically by detection of the presence of immunoglobulin (Ig)M antibodies or rising anti-HEV IgG titers. However, serological assays have demonstrated a significant variation in their sensitivities and specificities. In this study, we present the systematic comparison of different immunological anti-HEV assays using complete seroconversion panels of 10 virologically confirmed HEV genotype 3 infected individuals. Assay sensitivities were further evaluated by testing serially diluted World Health Organization (WHO) reference reagent for hepatitis E virus antibody and one patient sample infected with HEV genotype 3. Anti-HEV IgM and IgG antibody presence was determined using the immunological assays Wantai HEV IgM/IgG enzyme-linked immunosorbent assay (ELISA) (Sanbio, Uden, The Netherlands), recomWell HEV IgM/IgG (Mikrogen, Neuried, Germany), HEV IgM ELISA 3.0, HEV ELISA, HEV ELISA 4.0, Assure HEV IgM Rapid Test (all MP Biomedicals Europe, Illkirch Cedex, France) and Anti-HEV ELISA (IgM/IgG, Euroimmun, Lübeck, Germany). The assays showed differences regarding their analytical and diagnostic sensitivities, with anti-HEV IgM assays (n = 5) being more divergent compared to anti-HEV IgG (n = 4) assays in this study. Considerable variations were observed particularly for the detection period of IgM antibodies. This is the first study systematically characterizing serologic assays on the basis of seroconversion panels, providing sample conformity for a conclusive comparison. Future studies should include the assay comparison covering the four different genotypes. PMID:27556482

  9. [Obtention of a heterohybridoma for production of type IgM monoclonal antibodies against the D antigen of the Rh system].

    PubMed

    León-González, Graciela; Cruz, Carlos

    2007-03-01

    The objective was to obtain a heterohybridoma capable of producing a monoclonal antibody with IgM type anti-D specificity (Rh system), that could be used as a reactive for hemoclasification. Mononuclear cells (MNC) were extracted from a blood sample of a highly sensitized woman, five days after giving birth to an Rh positive child. These were then transformed with the culture supernatant (CSN) of B05.8 cells, rich in Epstein Barr virus (EBV). Once transformed and in exponential growth, they were fused with K6H6/B5 line cells using PEG 4.000 as a fusing agent in a 1:1 proportion. After fusion, they were seeded in culture plates in order to evaluate the formation of hybrids and the secretion of specific antibodies in the CSN of each well. The efficiency of the fusion was 1.8 x 10(-6), making it possible to obtain an anti-D IgM producing clone, which we named BMS-9. This clone could be maintained in constant culture for three months, producing antibodies in a concentration of 4 microg/mL in de CSN. It was also possible to obtain antibodies with an Artificial Capilar System (ACS) reaching a concentration of 24 microg/mL. Potency was determined using Ror cells. In CSN at immediate centrifugation (IC): 1 x 32, score 52; 15' from incubation at room temperature (RT): 1 x 1,024 score 105. With that ACS product at IC: 1 x 32 score 54; 15' from incubation at RT: 1 x 8.192 score 136; and a 37 degrees C: 1 x 8,192 score 136. Reactivity was detected with red cells D(IIIa), D(IV), D(Va), D(VI) type IV, D(VII), DFR, DNU, STEM+, DAR and DAU. There was no reactivity with red cells D(IIIc), DI(Va), D(V) type II, D(VI) types I, II y III, Ro(HAR), DOL and weak D type II. During field study, no false negative or false positive reactions were detected. A stable heterohybridoma was obtained, producer of IgM type anti-D, with enough qualities to be used in blood typing. Given the excellent qualities of the antibody, we are evaluating dilution media and the addition of type IgG antibodies

  10. Comparison of Three Commercially Available Serologic Assays Used To Detect Human Parvovirus B19-Specific Immunoglobulin M (IgM) and IgG Antibodies in Sera of Pregnant Women

    PubMed Central

    Butchko, Allyson R.; Jordan, Jeanne A.

    2004-01-01

    A split-sample study was conducted to evaluate the performances of three enzyme immunoassays (EIAs) utilizing one or more conformational antigens to detect human parvovirus B19 (B19V)-specific immunoglobulin M (IgM) or IgG in the sera of 198 pregnant women. We compared EIAs available from Biotrin International, Inc. (Dublin, Ireland), Medac Diagnostika (Wedel, Germany), and Mikrogen (Martinsried, Germany). Specimens with discordant results were analyzed further using an immunofluorescence assay (Biotrin). Equivocal data accounted for close to half of all the discrepant results for both IgM and IgG, with 7 of 15 discrepant results from the Medac and Mikrogen kits involving equivocal data and the Biotrin kit giving a single equivocal result. For each specimen, a consensus was established from the four test results if agreement occurred among at least three of four results. Overall, the highest percentage of agreement with the consensus results was seen when Biotrin kits were used; 194 (100%) of 194 and 194 (99.5%) of 195 results for IgM and IgG, respectively, agreed with the consensus results. When Medac kits were used, 189 (97.4%) of 194 and 191 (97.9%) of 195 results for IgM and IgG, respectively, agreed with the consensus, and when Mikrogen kits were used, 179 (92.3%) of 194 and 193 (99%) of 195 results for IgM and IgG, respectively, agreed with the consensus. Given the consensus results, the Medac EIA appeared to generate presumed false-positive results for IgM and the Mikrogen EIA appeared to generate presumed false-positive results for IgG and IgM. In summary, the Biotrin EIAs produced far fewer equivocal results than the other assays and results of the Biotrin EIAs agreed more often with the consensus results than did those of the other commercially available EIAs for detecting B19V-specific IgM and IgG antibodies. PMID:15243081

  11. Diffusion-in-gel enzyme-linked immunosorbent assay for routine detection of IgG and IgM antibodies to Campylobacter jejuni.

    PubMed

    Svedhem, A; Gunnarsson, H; Kaijser, B

    1983-07-01

    A diagnostic serologic test for antibodies to Campylobacter jejuni is presented. A diffusion-in-gel enzyme-linked immunosorbent assay with a surface antigen pool from two C jejuni strains was used. In testing serum samples for antibodies to C jejuni this system possessed high specificity, reliability, sensitivity, and capacity. It is easy to perform and there is no demand for expensive equipment. IgG and IgM antibodies can be quantitated separately, which allows for discrimination between individuals with acute infections and those with antibodies remaining after recovery. One serum sample is often sufficient to obtain such information. Seven groups of individuals were investigated. Serum samples from a total of 159 patients in three different groups and from 306 healthy individuals in four different groups were tested.

  12. Variable region structure and staphylococcal protein A binding specificity of a mouse monoclonal IgM anti-laminin-receptor antibody.

    PubMed Central

    Feijó, G C; Sabbaga, J; Carneiro, C R; Brígido, M M

    1997-01-01

    Staphylococcal protein A is a cell wall-attached polypeptide that acts as a B-lymphocyte superantigen. This activation correlates with specific VH gene segment usage in the B-cell receptor. B-cell receptor assembled from members of the VH3 family in humans, or S107 family in mice, has an intrinsic affinity for protein A. Human VH3-derived antibodies bind to domain D of protein A. We have characterized a mouse IgM monoclonal antibody that binds protein A. The sequencing of the variable region suggests an almost germline-encoded VH derived from S107 family and a V kappa 8-derived VL. The binding specificity of the monoclonal antibody was tested with various recombinant constructions derived from protein A. We show that, unlike human VH3-derived antibody, mouse S107-derived immunoglobulin binds to the B domain of the bacterial superantigen. PMID:9301540

  13. Molecular cloning of a new immunomodulatory protein from Anoectochilus formosanus which induces B cell IgM secretion through a T-independent mechanism.

    PubMed

    Kuan, Yen-Chou; Wu, Tsai-Jen; Kuo, Che-Yu; Hsu, Ju-Chun; Chang, Wen-Ying; Sheu, Fuu

    2011-01-01

    An immunomodulatory protein (IPAF) was purified and cloned from Anoectochilus formosanus, an Orchidaceae herbal plant in Asia. The major targeting immune cells of IPAF and its modulating effects toward B lymphocytes were investigated. Rapid amplification of cDNA ends (RACE) was conducted to clone the IPAF gene, and the obtained sequence was BLAST compared on the NCBI database. MACS-purified mouse T and B lymphocytes were stimulated with IPAF and the cell proliferation, activation, and Igs production were examined. IPAF comprised a 25 amino acids signal peptide and a 138 amino acids protein which was homologous to the lectins from Orchidaceae plant. IPAF selectively induced the cell proliferation in mouse splenic B lymphocytes but not T lymphocytes. The IPAF-induced B cells exhibited increased CD69 and MHC class II expression, and a dose- and time-dependent enhancement in IgM production. These results suggested potential benefits of IPAF to strengthen the humoral immunity. PMID:21698210

  14. The Use of NS1 Rapid Diagnostic Test and qRT-PCR to Complement IgM ELISA for Improved Dengue Diagnosis from Single Specimen.

    PubMed

    Teoh, Boon-Teong; Sam, Sing-Sin; Tan, Kim-Kee; Johari, Jefree; Abd-Jamil, Juraina; Hooi, Poh-Sim; AbuBakar, Sazaly

    2016-06-09

    Timely and accurate dengue diagnosis is important for differential diagnosis and immediate implementation of appropriate disease control measures. In this study, we compared the usefulness and applicability of NS1 RDT (NS1 Ag Strip) and qRT-PCR tests in complementing the IgM ELISA for dengue diagnosis on single serum specimen (n = 375). The NS1 Ag Strip and qRT-PCR showed a fair concordance (κ = 0.207, p = 0.001). While the NS1 Ag Strip showed higher positivity than qRT-PCR for acute (97.8% vs. 84.8%) and post-acute samples (94.8% vs. 71.8%) of primary infection, qRT-PCR showed higher positivity for acute (58.1% vs. 48.4%) and post-acute (50.0% vs.41.4%) samples in secondary infection. IgM ELISA showed higher positivity in samples from secondary dengue (74.2-94.8%) than in those from primary dengue (21.7-64.1%). More primary dengue samples showed positive with combined NS1 Ag Strip/IgM ELISA (99.0% vs. 92.8%) whereas more secondary samples showed positive with combined qRT-PCR/IgM ELISA (99.4% vs. 96.2%). Combined NS1 Ag Strip/IgM ELISA is a suitable combination tests for timely and accurate dengue diagnosis on single serum specimen. If complemented with qRT-PCR, combined NS1 Ag Strip/IgM ELISA would improve detection of secondary dengue samples.

  15. A potent neutralizing IgM mAb targeting the N218 epitope on E2 protein protects against Chikungunya virus pathogenesis

    PubMed Central

    Lam, Shirley; Nyo, Min; Phuektes, Patchara; Yew, Chow Wenn; Tan, Yee Joo; Chu, Justin Jang Hann

    2015-01-01

    Chikungunya virus (CHIKV) is a medically important human viral pathogen that causes Chikungunya fever accompanied with debilitating and persistent joint pain. Host-elicited or passively-transferred monoclonal antibodies (mAb) are essential mediators of CHIKV clearance. Therefore, this study aimed to generate and characterize a panel of mAbs for their neutralization efficacy against CHIKV infection in a cell-based and murine model. To evaluate their antigenicity and neutralization profile, indirect enzyme-linked immunosorbent assay (ELISA), an immunofluorescence assay (IFA) and a plaque reduction neutralization test were performed on mAbs of IgM isotype. CHIKV escape mutants against mAb 3E7b neutralization were generated, and reverse genetics techniques were then used to create an infectious CHIKV clone with a single mutation. 3E7b was also administered to neonate mice prior or after CHIKV infection. The survival rate, CHIKV burden in tissues and histopathology of the limb muscles were evaluated. Both IgM 3E7b and 8A2c bind strongly to native CHIKV surface and potently neutralize CHIKV replication. Further analyses of 3E7b binding and neutralization of CHIKV single-mutant clones revealed that N218 of CHIKV E2 protein is a potent neutralizing epitope. In a pre-binding neutralization assay, 3E7b blocks CHIKV attachment to permissive cells, possibly by binding to the surface-accessible E2-N218 residue. Prophylactic administration of 3E7b to neonate mice markedly reduced viremia and protected against CHIKV pathogenesis in various mice tissues. Given therapeutically at 4 h post-infection, 3E7b conferred 100% survival rate and similarly reduced CHIKV load in most mice tissues except the limb muscles. Collectively, these findings highlight the usefulness of 3E7b for future prophylactic or epitope-based vaccine design. PMID:26305993

  16. A potent neutralizing IgM mAb targeting the N218 epitope on E2 protein protects against Chikungunya virus pathogenesis.

    PubMed

    Lam, Shirley; Nyo, Min; Phuektes, Patchara; Yew, Chow Wenn; Tan, Yee Joo; Chu, Justin Jang Hann

    2015-01-01

    Chikungunya virus (CHIKV) is a medically important human viral pathogen that causes Chikungunya fever accompanied with debilitating and persistent joint pain. Host-elicited or passively-transferred monoclonal antibodies (mAb) are essential mediators of CHIKV clearance. Therefore, this study aimed to generate and characterize a panel of mAbs for their neutralization efficacy against CHIKV infection in a cell-based and murine model. To evaluate their antigenicity and neutralization profile, indirect enzyme-linked immunosorbent assay (ELISA), an immunofluorescence assay (IFA) and a plaque reduction neutralization test were performed on mAbs of IgM isotype. CHIKV escape mutants against mAb 3E7b neutralization were generated, and reverse genetics techniques were then used to create an infectious CHIKV clone with a single mutation. 3E7b was also administered to neonate mice prior or after CHIKV infection. The survival rate, CHIKV burden in tissues and histopathology of the limb muscles were evaluated. Both IgM 3E7b and 8A2c bind strongly to native CHIKV surface and potently neutralize CHIKV replication. Further analyses of 3E7b binding and neutralization of CHIKV single-mutant clones revealed that N218 of CHIKV E2 protein is a potent neutralizing epitope. In a pre-binding neutralization assay, 3E7b blocks CHIKV attachment to permissive cells, possibly by binding to the surface-accessible E2-N218 residue. Prophylactic administration of 3E7b to neonate mice markedly reduced viremia and protected against CHIKV pathogenesis in various mice tissues. Given therapeutically at 4 h post-infection, 3E7b conferred 100% survival rate and similarly reduced CHIKV load in most mice tissues except the limb muscles. Collectively, these findings highlight the usefulness of 3E7b for future prophylactic or epitope-based vaccine design.

  17. Poor Correlation between Pneumococcal IgG and IgM Titers and Opsonophagocytic Activity in Vaccinated Patients with Multiple Myeloma and Waldenstrom's Macroglobulinemia.

    PubMed

    Karlsson, Johanna; Roalfe, Lucy; Hogevik, Harriet; Zancolli, Marta; Andréasson, Björn; Goldblatt, David; Wennerås, Christine

    2016-04-01

    Patients with multiple myeloma and other B cell disorders respond poorly to pneumococcal vaccination. Vaccine responsiveness is commonly determined by measuring pneumococcal serotype-specific antibodies by enzyme-linked immunosorbent assay (ELISA), by a functional opsonophagocytosis assay (OPA), or by both assays. We compared the two methods in vaccinated elderly patients with multiple myeloma, Waldenstrom's macroglobulinemia, and monoclonal gammopathy of undetermined significance (MGUS). Postvaccination sera from 45 patients (n= 15 from each patient group) and 15 control subjects were analyzed by multiplexed OPA for pneumococcal serotypes 4, 6B, 14, and 23F, and the results were compared to IgG and IgM antibody titers measured by ELISA. While there were significant correlations between pneumococcal OPA and IgG titers for all serotypes among the control subjects (correlation coefficients [r] between 0.51 and 0.85), no significant correlations were seen for any of the investigated serotypes in the myeloma group (r= -0.18 to 0.21) or in the group with Waldenstrom's macroglobulinemia (borderline significant correlations for 2 of 4 serotypes). The MGUS group resembled the control group by having good agreement between the two test methods for 3 of 4 serotypes (r= 0.53 to 0.80). Pneumococcal postvaccination IgM titers were very low in the myeloma patients compared to the other groups and did not correlate with the OPA results. To summarize, our data indicate that ELISA measurements may overestimate antipneumococcal immunity in elderly subjects with B cell malignancies and that a functional antibody test should be used specifically for myeloma and Waldenstrom's macroglobulinemia patients. PMID:26912783

  18. Development and Characterization of Monoclonal Antibodies to Yellow Fever Virus and Application in Antigen Detection and IgM Capture Enzyme-Linked Immunosorbent Assay

    PubMed Central

    Adungo, Ferdinard; Kamau, David; Inoue, Shingo; Hayasaka, Daisuke; Posadas-Herrera, Guillermo; Sang, Rosemary; Mwau, Matilu

    2016-01-01

    Yellow fever (YF) is an acute hemorrhagic viral infection transmitted by mosquitoes in Africa and South America. The major challenge in YF disease detection and confirmation of outbreaks in Africa is the limited availability of reference laboratories and the persistent lack of access to diagnostic tests. We used wild-type YF virus sequences to generate recombinant envelope protein in an Escherichia coli expression system. Both the recombinant protein and sucrose gradient-purified YF vaccine virus 17D (YF-17D) were used to immunize BALB/c mice to generate monoclonal antibodies (MAbs). Eight MAbs were established and systematically characterized by indirect enzyme-linked immunosorbent assay (ELISA), Western blot analysis, and immunofluorescence assay (IFA). The established MAbs showed strong reactivity with wild-type YF virus and recombinant protein with no detectable cross-reactivity to dengue virus or Japanese encephalitis virus. Epitope mapping showed strong binding of three MAbs to amino acid positions 1 to 51, while two MAbs mapped to amino acid positions 52 to 135 of the envelope protein. The remaining three MAbs did not show reactivity to envelope fragments. The established MAbs exert no neutralization against wild-type YF and 17D viruses (titer of <10 for both strains). The applicability of MAbs 8H3 and 3F4 was further evaluated using IgM capture ELISA. A total of 49 serum samples were analyzed, among which 12 positive patient and vaccinee samples were correctly identified. Using serum samples that were 2-fold serially diluted, the IgM capture ELISA was able to detect all YF-positive samples. Furthermore, MAb-based antigen detection ELISA enabled the detection of virus in culture supernatants containing titers of about 1,000 focus-forming units. PMID:27307452

  19. Development and Characterization of Monoclonal Antibodies to Yellow Fever Virus and Application in Antigen Detection and IgM Capture Enzyme-Linked Immunosorbent Assay.

    PubMed

    Adungo, Ferdinard; Yu, Fuxun; Kamau, David; Inoue, Shingo; Hayasaka, Daisuke; Posadas-Herrera, Guillermo; Sang, Rosemary; Mwau, Matilu; Morita, Kouichi

    2016-08-01

    Yellow fever (YF) is an acute hemorrhagic viral infection transmitted by mosquitoes in Africa and South America. The major challenge in YF disease detection and confirmation of outbreaks in Africa is the limited availability of reference laboratories and the persistent lack of access to diagnostic tests. We used wild-type YF virus sequences to generate recombinant envelope protein in an Escherichia coli expression system. Both the recombinant protein and sucrose gradient-purified YF vaccine virus 17D (YF-17D) were used to immunize BALB/c mice to generate monoclonal antibodies (MAbs). Eight MAbs were established and systematically characterized by indirect enzyme-linked immunosorbent assay (ELISA), Western blot analysis, and immunofluorescence assay (IFA). The established MAbs showed strong reactivity with wild-type YF virus and recombinant protein with no detectable cross-reactivity to dengue virus or Japanese encephalitis virus. Epitope mapping showed strong binding of three MAbs to amino acid positions 1 to 51, while two MAbs mapped to amino acid positions 52 to 135 of the envelope protein. The remaining three MAbs did not show reactivity to envelope fragments. The established MAbs exert no neutralization against wild-type YF and 17D viruses (titer of <10 for both strains). The applicability of MAbs 8H3 and 3F4 was further evaluated using IgM capture ELISA. A total of 49 serum samples were analyzed, among which 12 positive patient and vaccinee samples were correctly identified. Using serum samples that were 2-fold serially diluted, the IgM capture ELISA was able to detect all YF-positive samples. Furthermore, MAb-based antigen detection ELISA enabled the detection of virus in culture supernatants containing titers of about 1,000 focus-forming units. PMID:27307452

  20. The Use of NS1 Rapid Diagnostic Test and qRT-PCR to Complement IgM ELISA for Improved Dengue Diagnosis from Single Specimen

    PubMed Central

    Teoh, Boon-Teong; Sam, Sing-Sin; Tan, Kim-Kee; Johari, Jefree; Abd-Jamil, Juraina; Hooi, Poh-Sim; AbuBakar, Sazaly

    2016-01-01

    Timely and accurate dengue diagnosis is important for differential diagnosis and immediate implementation of appropriate disease control measures. In this study, we compared the usefulness and applicability of NS1 RDT (NS1 Ag Strip) and qRT-PCR tests in complementing the IgM ELISA for dengue diagnosis on single serum specimen (n = 375). The NS1 Ag Strip and qRT-PCR showed a fair concordance (κ = 0.207, p = 0.001). While the NS1 Ag Strip showed higher positivity than qRT-PCR for acute (97.8% vs. 84.8%) and post-acute samples (94.8% vs. 71.8%) of primary infection, qRT-PCR showed higher positivity for acute (58.1% vs. 48.4%) and post-acute (50.0% vs.41.4%) samples in secondary infection. IgM ELISA showed higher positivity in samples from secondary dengue (74.2–94.8%) than in those from primary dengue (21.7–64.1%). More primary dengue samples showed positive with combined NS1 Ag Strip/IgM ELISA (99.0% vs. 92.8%) whereas more secondary samples showed positive with combined qRT-PCR/IgM ELISA (99.4% vs. 96.2%). Combined NS1 Ag Strip/IgM ELISA is a suitable combination tests for timely and accurate dengue diagnosis on single serum specimen. If complemented with qRT-PCR, combined NS1 Ag Strip/IgM ELISA would improve detection of secondary dengue samples. PMID:27278716

  1. A negative nontreponemal and/or specific antitreponemal IgM test does not exclude active infectious syphilis: evidence from a rabbit infectivity test

    PubMed Central

    Lin, Li-Rong; Tong, Man-Li; Gao, Kun; Zhu, Xiao-Zhen; Fan, Jin-Yi; Zheng, Wei-Hong; Li, Shu-Lian; Lin, Hui-Ling; Liu, Li-Li; Yang, Tian-Ci

    2016-01-01

    Abstract Background: The diagnostic criteria for active infectious syphilis in the clinic are important matter of controversy and debate. So far, clinicians habitually do use the negative results of the nontreponemal and/or the specific antitreponemal IgM as the evidences of disease-free or active infection-free status. Method: We present a case study involving a patient who was admitted to Zhongshan Hospital because of cerebral infarct. Clinical examination indicated he had a history of latent syphilis with negative nontreponemal and specific antitreponemal IgM tests. The cerebrospinal fluid sample from the patient was inoculated into seronegative New Zealand rabbit. Results: Motile Treponema pallidum was detected by a rabbit infectivity test in the patient's cerebrospinal fluid. This syphilis strain was confirmed by DNA subtyping form of “centers for disease control subtype/tp0548 sequence type”, and the strain type was 14d/f. Treatment with benzathine penicillin provided no apparent benefit, but treatment with aqueous crystalline penicillin G, especially recommended for neurosyphilis, led to disease regression. No evidence of cerebral infarct was observed during a 2-year follow-up period. Conclusion: The definitive differential diagnosis of active infectious syphilis should be reconsidered. Moreover, selecting the appropriate penicillin preparation is important because T pallidum can reside in sequestered sites. It is necessary to treat a patient with known invasion of the central nervous system with aqueous crystalline penicillin G, if previous treatment for syphilis failed and patients had some clinical neurological presentation that is otherwise unexplained, but that could represent neurosyphilis. Additional studies are needed to confirm the results in other syphilis patients. PMID:27495107

  2. Serum levels of immunoglobulins (IgG, IgA, IgM) in Antarctic summer expeditioners and their relationship with seasickness.

    PubMed

    Mishra, K P; Yadav, A P; Shweta; Chanda, Sudipta; Majumdar, D; Ganju, Lilly

    2011-01-01

    The Antarctic continent is full of environmental extremes like isolation, cold, UV exposure, and blizzards etc. The present study was conducted to analyze the effect of ship borne journey and the impact of Antarctic harsh environment on serum immunoglobulin (IgG, IgM, IgA) levels and their relationship with seasickness in Indian expeditioners. It was observed that one month onboard ship journey induced an increase in serum IgA levels and decrease in IgG levels while after being one month off board at the Indian research station Maitri, decreased levels of IgG and increased levels of IgA were found. IgM levels were not altered in comparison to the base line control. Moreover, serum IgG level showed a positive correlation while IgA level showed a negative correlation with seasickness. The stimulation of human peripheral blood mononuclear cells (PBMCs) with serum of expeditioner at different places showed that IgA at lower dose induces the release of pro-inflammatory IL-1β, and IL-6 cytokines from PBMCs while higher dose of IgA decreases proinflammatory cytokine production. The release of anti-inflammatory cytokines TGF-β1 and IL-10 was not significantly altered. Thus, the present study concluded that ship borne journey and Antarctic environment lead to increased serum IgA levels while decreased IgG levels. It also suggests that serum IgA level could be a possible biomarker for environmental stress.

  3. Therapeutics to promote CNS repair: a natural human neuron-binding IgM regulates membrane-raft dynamics and improves motility in a mouse model of multiple sclerosis.

    PubMed

    Xu, Xiaohua; Denic, Aleksandar; Warrington, Arthur E; Bieber, Allan J; Rodriguez, Moses

    2013-01-01

    We have discovered a role for natural autoantibodies in central nervous system repair, remyelination and axon protection. These natural human antibodies are of the immunoglobulin M (IgM) isotype, and they bind to the surface of neural cells. The epitope of the antibody includes sialic acid because treatment with sialidase disrupts the binding. A fully human recombinant form of one of these IgMs, rHIgM12, has the same properties as the serum-derived IgM. rHIgM12 enhanced polarized axonal outgrowth from primary neurons when presented as a substrate in vitro and improved motor functions in chronically Theiler's virus-infected SJL mice, a model of MS. rHIgM12 bound to neuronal surfaces and induced cholesterol and ganglioside (GM1) clustering, indicating that rHIgM12 functions through a mechanism of axonal membrane stabilization. Our work demonstrates that a natural human neuron-binding IgM can regulate membrane domain dynamics. This antibody has the potential to improve neurologic disease.

  4. Trends in the patterns of IgM and IgG antibodies in febrile persons with suspected dengue in Barbados, an English-speaking Caribbean country, 2006-2013.

    PubMed

    Kumar, Alok; Nielsen, Anders L

    2015-01-01

    Long-term seroprevalence studies of dengue have provided a measure of the degree of endemicity and future trends in disease prevalence and severity. In this study, we describe the seroprevalence of dengue antibodies in febrile persons with suspected acute dengue in Barbados. It is a retrospective population-based study of all febrile persons with suspected dengue from 2006 to 2013. All of the cases had IgM and IgG antibodies in the blood sample drawn between days 3 and 5 of their illness. Among the 8296 cases that were tested for IgM antibodies, 3037 (36.6%) had recent dengue infection. In the age groups <5 years, 5-20 years and >20 years, 23.3%, 39.6% and 35.5% had acute infection, respectively. Of the 7227 cases with documented IgG results, 5473 (75.7%) were positive and had a past infection. In the age groups <5 years, 5-20 years and >20 years, 31.2%, 65.2% and 86.6%, respectively, had a past infection (IgG positive). During the first 5 years of life, 10-20% of febrile persons investigated for dengue had a positive IgM and a negative IgG titer, between 5 and 10% had a positive IgM and IgG titer, 5% had a positive IgG and a negative IgM titer, and between 45% and 65% had a negative IgM and a negative IgG titer. Throughout the study period, between 12% and 20% of febrile persons failed to show any evidence of current or previous dengue. In the age groups <5 years, 5-20 years and >20 years, 45.0%, 18.8% and 7.2%, respectively, had no evidence of recent or past dengue (both IgM and IgG negative). Between 37% and 59% of the febrile persons had serological evidence of past dengue in the absence of any current dengue. In conclusion, the pattern of IgG antibodies in this study was comparable to those in countries known to be hyperendemic for dengue. The age of infection is likely to shift to younger adults and children who are more likely to have severe dengue in the future.

  5. A case of severe chronic inflammatory demyelinating polyradiculoneuropathy with monoclonal gammopathy of undetermined significance with alternating immunoglobulin class to IgM from IgA.

    PubMed

    Hayashi, Shintaro; Nagamine, Shun; Makioka, Kouki; Kusunoki, Susumu; Okamoto, Koichi

    2016-09-29

    A 71-year-old woman with chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) with IgA-λ monoclonal gammopathy of undetermined significance (MGUS) showed the acute development of tetraplegia, respiratory failure, and a marked fluctuation of the blood pressure. Intravenous (IV) high-dose steroid therapy (methylprednisolone: 1 g/day × 3 days), followed by oral prednisolone (PSL) (40 mg/day), and IV immunoglobulin (IVIg, 0.4 g/kg/day × 5 days) administrations resulted in the amelioration of these symptoms. However, they soon relapsed, which eventually led to complete tetraplegia and the need for mechanical ventilation. At this time, serum components of IgA-λ and IgM-λ were biclonally positive. Seven courses of plasma exchange and the alternative administration of dexamethasone (12 mg/day) and methtorexate (15 mg/week) were conducted, but with no significant improvement. Nine months after admission, she showed totally-locked in syndrome. Cryo-preserved serum obtained at this time showed high titers of IgM class antibodies against ganglioside (GD3 +++, GT1a ++++, GT1b ++, GQ1b +++, and GD1b +++), which had been negative on admission. Biopsy of the left sural nerve showed moderate reductions of large and small myelinated fibers with no inflammation, no depositions of amyloid, IgG, IgA, or IgM, and teased fiber findings revealed neither myelin ovoids nor segmental demyelination. Alternatively, melphalan at 5 mg and PSL at 32 mg were administered, with no amelioration, while serum IgA-λ monoclonal protein diminished, and IgM-λ M protein positivity was continuously observed. She frequently developed sepsis; therefore, we could no longer continue any immunosupressive therapies, but monthly IVIg administrations were given. Twelve months after admission, her neurological symptoms gradually improved and she was weaned off of mechanical ventilation. Eighteen months after admission, her muscle strength corresponded to 2 on manual muscle testing

  6. A case of severe chronic inflammatory demyelinating polyradiculoneuropathy with monoclonal gammopathy of undetermined significance with alternating immunoglobulin class to IgM from IgA.

    PubMed

    Hayashi, Shintaro; Nagamine, Shun; Makioka, Kouki; Kusunoki, Susumu; Okamoto, Koichi

    2016-09-29

    A 71-year-old woman with chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) with IgA-λ monoclonal gammopathy of undetermined significance (MGUS) showed the acute development of tetraplegia, respiratory failure, and a marked fluctuation of the blood pressure. Intravenous (IV) high-dose steroid therapy (methylprednisolone: 1 g/day × 3 days), followed by oral prednisolone (PSL) (40 mg/day), and IV immunoglobulin (IVIg, 0.4 g/kg/day × 5 days) administrations resulted in the amelioration of these symptoms. However, they soon relapsed, which eventually led to complete tetraplegia and the need for mechanical ventilation. At this time, serum components of IgA-λ and IgM-λ were biclonally positive. Seven courses of plasma exchange and the alternative administration of dexamethasone (12 mg/day) and methtorexate (15 mg/week) were conducted, but with no significant improvement. Nine months after admission, she showed totally-locked in syndrome. Cryo-preserved serum obtained at this time showed high titers of IgM class antibodies against ganglioside (GD3 +++, GT1a ++++, GT1b ++, GQ1b +++, and GD1b +++), which had been negative on admission. Biopsy of the left sural nerve showed moderate reductions of large and small myelinated fibers with no inflammation, no depositions of amyloid, IgG, IgA, or IgM, and teased fiber findings revealed neither myelin ovoids nor segmental demyelination. Alternatively, melphalan at 5 mg and PSL at 32 mg were administered, with no amelioration, while serum IgA-λ monoclonal protein diminished, and IgM-λ M protein positivity was continuously observed. She frequently developed sepsis; therefore, we could no longer continue any immunosupressive therapies, but monthly IVIg administrations were given. Twelve months after admission, her neurological symptoms gradually improved and she was weaned off of mechanical ventilation. Eighteen months after admission, her muscle strength corresponded to 2 on manual muscle testing

  7. Immunoglobulin (Ig)M antibodies against oxidized cardiolipin but not native cardiolipin are novel biomarkers in haemodialysis patients, associated negatively with mortality

    PubMed Central

    Frostegård, A G; Hua, X; Su, J; Carrero, J J; Heimbürger, O; Bárány, P; Stenvinkel, P; Frostegård, J

    2013-01-01

    The risk of premature death is high in haemodialysis (HD) patients. Antibodies against cardiolipin (anti-CL) are thrombogenic in diseases such as systemic lupus erythematosus (SLE). CL is easily oxidized (Ox) and plays a role in apoptosis. In this work we studied immunoglobulin (Ig)M anti-CL and anti-OxCL in HD-patients. We conducted an observational study with a prospective follow-up examining the relationship between anti-CL, anti-OxCL and mortality risk in a well-characterized cohort of 221 prevalent HD patients [56% men, median age 66 (interquartile range 51–74) years, vintage time 29 (15–58) months] with a mean follow-up period of 41 (20–48 months). According to the receiver operator characteristic (ROC) analysis, anti-OxCL [area under the curve (AUC) 0·62, P < 0·01], but not anti-CL (AUC 0·52, P = 0·2), is associated with mortality. In crude and adjusted Cox analysis, every log increase in anti-OxCL inversely predicted all-cause [adjusted hazard ratios (HR) 0·62 (0·43–0·89)] and CVD-related [adjusted HR 0·56 (0·32–0·98)] mortality. Patients with anti-OxCL levels below median also had increased all-cause and cardiovascular disease (CVD)-related mortality. Although anti-OxCL and anti-phosphorylcholine (PC) were related positively to each other (ρ = 0·57, P < 0·01), patients with one or two of these autoantibody levels below the median were associated with an incrementally increased death risk. Anti-OxCL were co-factor β2-GPI-independent; anti-CL from patients with anti-phospholipid antibody syndrome were β2-GPI-dependent, while sera from HD-patients less so. Sera from healthy donors was not β2-GPI-dependent. Anti-OxCL IgM is β2-glycoprotein 1 (GPI)-independent and a novel biomarker; low levels are associated with death among HD patients (and high levels with decreased risk). Combination with anti-PC increases this association. Putative therapeutic implications warrant further investigation. PMID:23879320

  8. Development of a lateral flow immunochromatographic assay for rapid detection of Mycoplasma pneumoniae-specific IgM in human serum specimens.

    PubMed

    Ou, Liming; Lv, Qingyu; Wu, Canjun; Hao, Huaijie; Zheng, Yuling; Jiang, Yongqiang

    2016-05-01

    Early diagnosis of Mycoplasma pneumoniae (MP) infection is crucial for prompt treatment and good patient outcome. However, serological tests to detect MP rapidly and conveniently are still lacking. This study aimed to use the fluorescent dye Alexa Fluor® 647 as the detection marker to develop a lateral flow immunochromatographic assay (LFIA) for detection of MP-specific IgM in serum specimen. Monoclonal mouse antibody against human IgM (μ-chain specific) and goat anti-rabbit IgG were labeled with Alexa Fluor® 647 (anti-IgM-AF647 and anti-IgG-AF647). A mixture of natural MP antigen and recombinant P1 antigen was coated as the test line (T line) and rabbit IgG was coated as the control line (C line) on a nitrocellulose (NC) membrane. The MP antigens captured IgM-anti-IgM-AF647 complex on the T line. Rabbit IgG captured anti-IgG-AF647 on the C line. The fluorescence intensity on the T line and C line was measured. Sartorius CN140 NC membrane showed higher sensitivity than CN95. The optimal reaction time for the LFIA was 10min. The area under the receiver operating characteristic curve based on 34 MP positive and 166 MP negative serum samples was 0.986 (p<0.001). The cutoff value of T/C area ratio was 0.3830. The LFIA strips did not react with serum from patients infected with non-MP pathogens including influenza viruses and bacteria causing respiratory tract infection. The intra-assay and inter-assay coefficients of variation were between 3.28% and 10.14%. The shelf life was calculated to be 2years at room temperature. The LFIA strips and the commercial EUROIMMUN kit showed consistent results on 372 serum specimens. The overall consistency rate was 96.37% with a Kappa value of 0.842 (p<0.001). The LFIA in the current study may be a sensitive and specific approach to detect early MP infection rapidly and conveniently. PMID:26979644

  9. Development of a lateral flow immunochromatographic assay for rapid detection of Mycoplasma pneumoniae-specific IgM in human serum specimens.

    PubMed

    Ou, Liming; Lv, Qingyu; Wu, Canjun; Hao, Huaijie; Zheng, Yuling; Jiang, Yongqiang

    2016-05-01

    Early diagnosis of Mycoplasma pneumoniae (MP) infection is crucial for prompt treatment and good patient outcome. However, serological tests to detect MP rapidly and conveniently are still lacking. This study aimed to use the fluorescent dye Alexa Fluor® 647 as the detection marker to develop a lateral flow immunochromatographic assay (LFIA) for detection of MP-specific IgM in serum specimen. Monoclonal mouse antibody against human IgM (μ-chain specific) and goat anti-rabbit IgG were labeled with Alexa Fluor® 647 (anti-IgM-AF647 and anti-IgG-AF647). A mixture of natural MP antigen and recombinant P1 antigen was coated as the test line (T line) and rabbit IgG was coated as the control line (C line) on a nitrocellulose (NC) membrane. The MP antigens captured IgM-anti-IgM-AF647 complex on the T line. Rabbit IgG captured anti-IgG-AF647 on the C line. The fluorescence intensity on the T line and C line was measured. Sartorius CN140 NC membrane showed higher sensitivity than CN95. The optimal reaction time for the LFIA was 10min. The area under the receiver operating characteristic curve based on 34 MP positive and 166 MP negative serum samples was 0.986 (p<0.001). The cutoff value of T/C area ratio was 0.3830. The LFIA strips did not react with serum from patients infected with non-MP pathogens including influenza viruses and bacteria causing respiratory tract infection. The intra-assay and inter-assay coefficients of variation were between 3.28% and 10.14%. The shelf life was calculated to be 2years at room temperature. The LFIA strips and the commercial EUROIMMUN kit showed consistent results on 372 serum specimens. The overall consistency rate was 96.37% with a Kappa value of 0.842 (p<0.001). The LFIA in the current study may be a sensitive and specific approach to detect early MP infection rapidly and conveniently.

  10. Cytotoxic activity against human neuroblastoma and melanoma cells mediated by IgM antibodies derived from peripheral blood of healthy donors.

    PubMed

    Devarapu, Satish Kumar; Mamidi, Srinivas; Plöger, Frank; Dill, Othmar; Blixt, Ola; Kirschfink, Michael; Schwartz-Albiez, Reinhard

    2016-06-15

    A small percentage of healthy donors identified in the Western population carry antibodies in their peripheral blood which convey cytotoxic activity against certain human melanoma and neuroblastoma cell lines. We measured the cytotoxic activity of sera and plasmas from healthy donors on the human neuroblastoma cell line Kelly and various melanoma cell lines. Antibodies of IgM isotype, presumably belonging to the class of naturally occurring antibodies, exerted cytotoxic activity in a complement-dependent fashion. Apart from complement-dependent tumor cell lysis, we observed C3 opsonization in all tumor cell lines upon treatment with cytotoxic plasmas. Cell lines tested primarily expressed membrane complement regulatory proteins (mCRP) CD46, CD55 and CD59 to various extents. Blocking of mCRPs by monoclonal antibodies enhanced cell lysis and opsonization, though some melanoma cells remained resistant to complement attack. Epitopes recognized by cytotoxic antibodies were represented by gangliosides such as GD2 and GD3, as evidenced by cellular sialidase pretreatment and enhanced expression of distinct gangliosides. It remains to be clarified why only a small fraction of healthy persons carry these antitumor cytotoxic antibodies. PMID:26830059

  11. Cytotoxic activity against human neuroblastoma and melanoma cells mediated by IgM antibodies derived from peripheral blood of healthy donors.

    PubMed

    Devarapu, Satish Kumar; Mamidi, Srinivas; Plöger, Frank; Dill, Othmar; Blixt, Ola; Kirschfink, Michael; Schwartz-Albiez, Reinhard

    2016-06-15

    A small percentage of healthy donors identified in the Western population carry antibodies in their peripheral blood which convey cytotoxic activity against certain human melanoma and neuroblastoma cell lines. We measured the cytotoxic activity of sera and plasmas from healthy donors on the human neuroblastoma cell line Kelly and various melanoma cell lines. Antibodies of IgM isotype, presumably belonging to the class of naturally occurring antibodies, exerted cytotoxic activity in a complement-dependent fashion. Apart from complement-dependent tumor cell lysis, we observed C3 opsonization in all tumor cell lines upon treatment with cytotoxic plasmas. Cell lines tested primarily expressed membrane complement regulatory proteins (mCRP) CD46, CD55 and CD59 to various extents. Blocking of mCRPs by monoclonal antibodies enhanced cell lysis and opsonization, though some melanoma cells remained resistant to complement attack. Epitopes recognized by cytotoxic antibodies were represented by gangliosides such as GD2 and GD3, as evidenced by cellular sialidase pretreatment and enhanced expression of distinct gangliosides. It remains to be clarified why only a small fraction of healthy persons carry these antitumor cytotoxic antibodies.

  12. Complete amino acid sequence of the variable domains of two human IgM anti-gamma globulins (Lay/Pom) with shared idiotypic specificities.

    PubMed

    Capra, J D; Klapper, D G

    1976-01-01

    On the basis of extensive shared idiotypic specificities, two human IgM anti-gamma-globulins (Lay/Pom) were selected for complete amino acid sequence analysis of their variable domains. Previous studies on the variable regions of the heavy chains of these proteins had shown but eight amino acid differences, only one of which was within a complementarity-determining hypervariable region. The complete amino acid sequence of the variable regions of the light chains of these two proteins is the subject of this report. Protein Lay is a typical VchiI protein with only five 'framework' differences when compared with protein Roy. Protein Pom is best classified as a VchiII, but in the 'framework' there are 16 differences between it and protein Ti. Although there are extensive differences in the first hypervariable region, the second and third light-chain hypervariable regions have an identical sequence. The finding of two identical light-chain and two identical heavy-chain hypervariable regions in these two proteins, which were selected on the basis of their combining specificities and their idiotypic cross-reactions, strongly implicates hypervariable regions in the constitution of the idiotypic determinants and the antibody combining site. Additionally, the finding of identical hypervariable regions in light chains of different V-region subgroups fulfills a prediction of the gene-interaction concept of antibody variability. PMID:824717

  13. Adjuvant Effect of Quillaja saponaria Saponin (QSS) on Protective Efficacy and IgM Generation in Turbot (Scophthalmus maximus) upon Immersion Vaccination

    PubMed Central

    Wang, Yujuan; Wang, Xiuhua; Huang, Jie; Li, Jun

    2016-01-01

    The adjuvant effect of Quillaja saponaria saponin (QSS) on protection of turbot fry was investigated with immersion vaccination of formalin-killed Vibrio anguillarum O1 and various concentrations of QSS (5, 25, 45 and 65 mg/L). Fish were challenged at days 7, 14 and 28 post-vaccination. Significantly high relative percent of survival (RPS) ((59.1 ± 13.6)%, (81.7 ± 8.2)%, (77.8 ± 9.6)%) were recorded in the fish that received bacterins immersion with QSS at 45 mg/L, which is comparable to the positive control group vaccinated by intraperitoneal injection (IP). Moreover, a remarkably higher serum antibody titer was also demonstrated after 28 days in the vaccinated fish with QSS (45 mg/L) than those vaccinated fish without QSS (p < 0.05), but lower than the IP immunized fish (p < 0.05). Significant upregulation of IgM gene expression has also been identified in the tissues of skin, gill, spleen and kidney from the immunized fish in comparison to the control fish. Taken together, the present study indicated that QSS was able to dramatically evoke systemic and mucosal immune responses in immunized fish. Therefore, QSS might be a promising adjuvant candidate for fish vaccination via an immersion administering route. PMID:26950114

  14. Immunohistochemical detection of IgM and IgG in lung tissue of dogs with leptospiral pulmonary haemorrhage syndrome (LPHS).

    PubMed

    Schuller, Simone; Callanan, John J; Worrall, Sheila; Francey, Thierry; Schweighauser, Ariane; Kohn, Barbara; Klopfleisch, Robert; Posthaus, Horst; Nally, Jarlath E

    2015-06-01

    Leptospiral pulmonary haemorrhage syndrome (LPHS) is a severe form of leptospirosis. Pathogenic mechanisms are poorly understood. Lung tissues from 26 dogs with LPHS, 5 dogs with pulmonary haemorrhage due to other causes and 6 healthy lungs were labelled for IgG (n=26), IgM (n=25) and leptospiral antigens (n=26). Three general staining patterns for IgG/IgM were observed in lungs of dogs with LPHS with most tissues showing more than one staining pattern: (1) alveolar septal wall staining, (2) staining favouring alveolar surfaces and (3) staining of intra-alveolar fluid. Healthy control lung showed no staining, whereas haemorrhagic lung from dogs not infected with Leptospira showed staining of intra-alveolar fluid and occasionally alveolar septa. Leptospiral antigens were not detected. We conclude that deposition of IgG/IgM is demonstrable in the majority of canine lungs with naturally occurring LPHS, similar to what has been described in other species. Our findings suggest involvement of the host humoral immunity in the pathogenesis of LPHS and provide further evidence to support the dog as a natural disease model for human LPHS. PMID:25963899

  15. Isolation of 1E4 IgM Anti-Fasciola hepatica Rediae Monoclonal Antibody from Ascites: Comparison of Two Purification Protocols.

    PubMed

    Alba, Annia; Marcet, Ricardo; Otero, Oscar; Hernández, Hilda M; Figueredo, Mabel; Sarracent, Jorge

    2016-02-01

    Purification of immunoglobulin M (IgM) antibodies could be challenging, and is often characterized by the optimization of the purification protocol to best suit the particular features of the molecule. Here, two different schemes were compared to purify, from ascites, the 1E4 IgM monoclonal antibody (mAb) previously raised against the stage of redia of the trematode Fasciola hepatica. This immunoglobulin is used as capture antibody in an immunoenzymatic assay to detect parasite ongoing infection in its intermediate hosts. The first purification protocol of the 1E4 mAb involved two chromatographic steps: an affinity chromatography on a Concanavalin A matrix followed by size exclusion chromatography. An immunoaffinity chromatography was selected as the second protocol for one-step purification of the antibody using the crude extract of adult parasites coupled to a commercial matrix. Immunoreactivity of the fractions during purification schemes was assessed by indirect immunoenzymatic assays against the crude extract of F. hepatica rediae, while purity was estimated by protein electrophoresis. Losses on the recovery of the antibody isolated by the first purification protocol occurred due to protein precipitation during the concentration of the sample and to low resolution of the size exclusion molecular chromatography step regarding this particular immunoglobulin. The immunoaffinity chromatography using F. hepatica antigens as ligands proved to be the most suitable protocol yielding a pure and immunoreactive antibody. The purification protocols used are discussed regarding efficiency and difficulties. PMID:26828226

  16. Phenotypic modulation of chronic lymphocytic leukemia cells by phorbol ester: induction of IgM secretion and changes in the expression of B cell-associated surface antigens.

    PubMed

    Gordon, J; Mellstedt, H; Aman, P; Biberfeld, P; Klein, G

    1984-01-01

    Freshly explanted neoplastic populations from 22 cases of phenotypically well-characterized chronic type B lymphocytic leukemia were studied for their capacity to respond to the phorbol ester TPA in vitro. In all but four cases the secretion of IgM was either induced or increased, often to a high level. In contrast, the export of free immunoglobulin (Ig) light chains, an almost consistent feature of the B lymphocytic leukemias, remained relatively constant after TPA treatment. Parallel changes in leukemic cell surface phenotype were probed with both "conventional" and monoclonal antibodies, revealing some modulation of markers in every case investigated. A diminution in the level of surface Ig (preferentially IgD) and the accumulation of cytoplasmic Ig observed after phorbol ester treatment were accompanied by a corresponding reduction or loss of the B1 antigen and usually of B2 when present. The most consistent change induced by TPA was the appearance of BB-1, a marker of activated B lymphocytes, which was rarely expressed on fresh leukemic cells. Another marker of activated lymphocytes, LB-1, was also often induced or increased in its expression after exposure of the cells to TPA. The magnitude of the TPA response appeared to relate to the stage of maturation arrest of the individual leukemic clones rather than to any clinical parameter explored. The significance of the findings to normal B cell differentiation and their potential clinical utility are discussed.

  17. RE-186 labeled 16.88 IgM and 88BV59 IgG human antibody studies to assess potential for radioimmunotherapy

    SciTech Connect

    Breitz, H.; Seiler, C.; Weiden, P. ||

    1994-05-01

    Two studies with Re-186-MAG{sub 2}GABA labeled human antibodies were carried out to assess feasibility for radioimmunotherapy. Antibodies 16.88 and 88BV59 react with different epitopes of CTA 16.88, a tumor associated antigen of colorectal carcinoma. In a phase I dose escalation study, 14 patients received 60 mg/m{sup 2} 16.88 IgM MoAb. The dose of Re-186 ranged from 25 mCi/m{sup 2} to 210 mCi/m{sup 2} divided into 3 weekly infusions. In a pilot study with 88BV59, a human IgG3k MoAb, 20 mg antibody was labeled with 25 mCi/m{sup 2} Re-186 and administered to 4 patients with colon carcinoma. Tumor targeting was seen in 12 of 14 patients with 16.88 and all 4 patients with 88BV59. Retention of antibody at the tumor was longer with 88BV59. One patient developed a rash. No other acute or delayed toxicities were observed. Human anti-human antibody did not develop in any patient. The slower metabolism of the 88BV59 IgG suggests that this form of immunoconjugate merits further investigation for use in radioimmunotherapy.

  18. Adjuvant Effect of Quillaja saponaria Saponin (QSS) on Protective Efficacy and IgM Generation in Turbot (Scophthalmus maximus) upon Immersion Vaccination.

    PubMed

    Wang, Yujuan; Wang, Xiuhua; Huang, Jie; Li, Jun

    2016-01-01

    The adjuvant effect of Quillaja saponaria saponin (QSS) on protection of turbot fry was investigated with immersion vaccination of formalin-killed Vibrio anguillarum O1 and various concentrations of QSS (5, 25, 45 and 65 mg/L). Fish were challenged at days 7, 14 and 28 post-vaccination. Significantly high relative percent of survival (RPS) ((59.1 ± 13.6)%, (81.7 ± 8.2)%, (77.8 ± 9.6)%) were recorded in the fish that received bacterins immersion with QSS at 45 mg/L, which is comparable to the positive control group vaccinated by intraperitoneal injection (IP). Moreover, a remarkably higher serum antibody titer was also demonstrated after 28 days in the vaccinated fish with QSS (45 mg/L) than those vaccinated fish without QSS (p < 0.05), but lower than the IP immunized fish (p < 0.05). Significant upregulation of IgM gene expression has also been identified in the tissues of skin, gill, spleen and kidney from the immunized fish in comparison to the control fish. Taken together, the present study indicated that QSS was able to dramatically evoke systemic and mucosal immune responses in immunized fish. Therefore, QSS might be a promising adjuvant candidate for fish vaccination via an immersion administering route. PMID:26950114

  19. Flow cytometry-based algorithm to analyze the anti-fixed Toxoplasma gondii tachyzoites IgM and IgG reactivity and diagnose human acute toxoplasmosis.

    PubMed

    Silva-dos-Santos, Priscila Pinto; Barros, Geisa Baptista; Mineo, José Roberto; de Oliveira Silva, Deise Aparecida; Menegaz, Mauro Hygino Weinert; Serufo, José Carlos; Dietze, Reynaldo; Martins-Filho, Olindo de Assis; Lemos, Elenice Moreira

    2012-04-30

    In the present study we evaluated the performance of a flow cytometry-based algorithm as a new serological approach to detect antibodies to T. gondii and specific IgG avidity to diagnose acute toxoplasmosis. The results showed that using FC-AFTA-IgM assay, all serum samples from patients with acute toxoplasmosis demonstrated seropositivity, whereas 90% of patients with chronic infection and 100% of non-infected individuals presented negative results. Thus, only 10% of patients with chronic toxoplasmosis showed residual IgM, in contrast with other methodologies used to diagnosis acute toxoplasmosis. On the order hand, FC-AFTA-IgG assay as well as FC-AFTA-IgG subclasses is unlikely to discriminate acute from chronic toxoplasmosis. We have also evaluated the performance of FC-AFTA-IgG avidity as a tool to exclude chronic toxoplasmosis in patients with positive FC-AFTA-IgM. Our data showed an excellent performance of FC-AFTA-IgG avidity employing the cut-off of 60% for Avidity Index (AI) with sensitivity and specificity of 100%. All serum samples from patients presenting acute toxoplasmosis showed low avidity index (AI≤60%), whereas all chronic patients showed high avidity index (AI>60%). The outstanding performance indexes of this novel flow cytometry-based algorithm support its use as a non-conventional alternative serological approach to diagnose human acute toxoplasmosis.

  20. Isolation of 1E4 IgM Anti-Fasciola hepatica Rediae Monoclonal Antibody from Ascites: Comparison of Two Purification Protocols.

    PubMed

    Alba, Annia; Marcet, Ricardo; Otero, Oscar; Hernández, Hilda M; Figueredo, Mabel; Sarracent, Jorge

    2016-02-01

    Purification of immunoglobulin M (IgM) antibodies could be challenging, and is often characterized by the optimization of the purification protocol to best suit the particular features of the molecule. Here, two different schemes were compared to purify, from ascites, the 1E4 IgM monoclonal antibody (mAb) previously raised against the stage of redia of the trematode Fasciola hepatica. This immunoglobulin is used as capture antibody in an immunoenzymatic assay to detect parasite ongoing infection in its intermediate hosts. The first purification protocol of the 1E4 mAb involved two chromatographic steps: an affinity chromatography on a Concanavalin A matrix followed by size exclusion chromatography. An immunoaffinity chromatography was selected as the second protocol for one-step purification of the antibody using the crude extract of adult parasites coupled to a commercial matrix. Immunoreactivity of the fractions during purification schemes was assessed by indirect immunoenzymatic assays against the crude extract of F. hepatica rediae, while purity was estimated by protein electrophoresis. Losses on the recovery of the antibody isolated by the first purification protocol occurred due to protein precipitation during the concentration of the sample and to low resolution of the size exclusion molecular chromatography step regarding this particular immunoglobulin. The immunoaffinity chromatography using F. hepatica antigens as ligands proved to be the most suitable protocol yielding a pure and immunoreactive antibody. The purification protocols used are discussed regarding efficiency and difficulties.

  1. Skin Injuries Reduce Survival and Modulate Corticosterone, C-Reactive Protein, Complement Component 3, IgM, and Prostaglandin E2 after Whole-Body Reactor-Produced Mixed Field (n + γ-Photons) Irradiation

    PubMed Central

    Kiang, Juliann G.; Ledney, G. David

    2013-01-01

    Skin injuries such as wounds or burns following whole-body γ-irradiation (radiation combined injury (RCI)) increase mortality more than whole-body γ-irradiation alone. Wound-induced decreases in survival after irradiation are triggered by sustained activation of inducible nitric oxide synthase pathways, persistent alteration of cytokine homeostasis, and increased susceptibility to systemic bacterial infection. Among these factors, radiation-induced increases in interleukin-6 (IL-6) concentrations in serum were amplified by skin wound trauma. Herein, the IL-6-induced stress proteins including C-reactive protein (CRP), complement 3 (C3), immunoglobulin M (IgM), and prostaglandin E2 (PGE2) were evaluated after skin injuries given following a mixed radiation environment that might be found after a nuclear incident. In this report, mice received 3 Gy of reactor-produced mixed field (n + γ-photons) radiations at 0.38 Gy/min followed by nonlethal skin wounding or burning. Both wounds and burns reduced survival and increased CRP, C3, and PGE2 in serum after radiation. Decreased IgM production along with an early rise in corticosterone followed by a subsequent decrease was noted for each RCI situation. These results suggest that RCI-induced alterations of corticosterone, CRP, C3, IgM, and PGE2 cause homeostatic imbalance and may contribute to reduced survival. Agents inhibiting these responses may prove to be therapeutic for RCI and improve related survival. PMID:24175013

  2. Aggregation of Streptococcus pneumoniae by a pneumococcal capsular polysaccharide-specific human monoclonal IgM correlates with antibody efficacy in vivo.

    PubMed

    Fabrizio, Kevin; Manix, Catherine; Guimaraes, Allan J; Nosanchuk, Joshua D; Pirofski, Liise-Anne

    2010-05-01

    Acquired antibody immunity to Streptococcus pneumoniae (pneumococcus) has been linked to serotype (ST)-specific opsonic antibodies to the relevant pneumococcal capsular polysaccharide (PPS) that mediate protection by enhancing the bactericidal effect of host phagocytes. Despite the well-recognized role of opsonic IgG in host defense against pneumococcus, PPS-specific monoclonal antibodies (MAbs) that mediate protection against lethal challenge with ST3 pneumococcus in mice but do not promote phagocytic killing in vitro (nonopsonic antibodies) have been described. In this study, we sought to determine the biological activity of one such MAb, A7 (a human PPS3-specific IgM), and the mechanism by which it mediates protection. In vitro studies demonstrated that coincubation of A7 with ST3 in the absence of phagocytes or a complement source resulted in a reduction in CFU on blood agar plates that was largely reversible by sonication. A chromogenic cellular proliferation assay demonstrated that A7 did not affect replication of ST3 in liquid culture. The ability of A7 to induce aggregation of ST3 was confirmed by fluorescence microscopy and flow cytometry: A7 induced aggregation of ST3, and in the presence of a complement source, A7 promoted deposition of complement component 3 (C3) on aggregated bacteria in a dose-dependent fashion. Similarly, administration of preincubated mixtures of A7 and ST3 intraperitoneally to mice protected them from the lethality of ST3 in a dose-dependent fashion. These findings suggest that A7-mediated aggregation enhances resistance to ST3, most likely by enhancing C3 deposition on the ST3 capsule, thereby promoting host antipneumococcal activity in vivo.

  3. DRB1*03:01 Haplotypes: Differential Contribution to Multiple Sclerosis Risk and Specific Association with the Presence of Intrathecal IgM Bands

    PubMed Central

    Cénit, M. Carmen; Urcelay, Elena; Arroyo, Rafael; Fernández, Óscar; Álvarez-Cermeño, José C.; Leyva, Laura

    2012-01-01

    Background Multiple sclerosis (MS) is a multifactorial disease with a genetic basis. The strongest associations with the disease lie in the Human Leukocyte Antigen (HLA) region. However, except for the DRB1*15:01 allele, the main risk factor associated to MS so far, no consistent effect has been described for any other variant. One example is HLA-DRB1*03:01, with a heterogeneous effect across populations and studies. We postulate that those discrepancies could be due to differences in the diverse haplotypes bearing that allele. Thus, we aimed at studying the association of DRB1*03:01 with MS susceptibility considering this allele globally and stratified by haplotypes. We also evaluated the association with the presence of oligoclonal IgM bands against myelin lipids (OCMB) in cerebrospinal fluid. Methods Genotyping of HLA-B, -DRB1 and -DQA1 was performed in 1068 MS patients and 624 ethnically matched healthy controls. One hundred and thirty-nine MS patients were classified according to the presence (M+, 58 patients)/absence (M−, 81 patients) of OCMB. Comparisons between groups (MS patients vs. controls and M+ vs. M−) were performed with the chi-square test or the Fisher exact test. Results Association of DRB1*03:01 with MS susceptibility was observed but with different haplotypic contribution, being the ancestral haplotype (AH) 18.2 the one causing the highest risk. Comparisons between M+, M− and controls showed that the AH 18.2 was affecting only M+ individuals, conferring a risk similar to that caused by DRB1*15:01. Conclusions The diverse DRB1*03:01-containing haplotypes contribute with different risk to MS susceptibility. The AH 18.2 causes the highest risk and affects only to individuals showing OCMB. PMID:22363536

  4. A new fluorescence/PET probe for targeting intracellular human telomerase reverse transcriptase (hTERT) using Tat peptide-conjugated IgM.

    PubMed

    Jung, Kyung Oh; Youn, Hyewon; Kim, Seung Hoo; Kim, Young-Hwa; Kang, Keon Wook; Chung, June-Key

    2016-08-26

    Despite an increasing need for methods to visualize intracellular proteins in vivo, the majority of antibody-based imaging methods available can only detect membrane proteins. The human telomerase reverse transcriptase (hTERT) is an intracellular target of great interest because of its high expression in several types of cancer. In this study, we developed a new probe for hTERT using the Tat peptide. An hTERT antibody (IgG or IgM) was conjugated with the Tat peptide, a fluorescence dye and (64)Cu. HT29 (hTERT+) and U2OS (hTERT-) were used to visualize the intracellular hTERT. The hTERT was detected by RT-PCR and western blot. Fluorescence signals for hTERT were obtained by confocal microscopy, live cell imaging, and analyzed by Tissue-FAXS. In nude mice, tumors were visualized using the fluorescence imaging devices Maestro™ and PETBOX. In RT-PCR and western blot, the expression of hTERT was detected in HT29 cells, but not in U2OS cells. Fluorescence signals were clearly observed in HT29 cells and in U2OS cells after 1 h of treatment, but signals were only detected in HT29 cells after 24 h. Confocal microscopy showed that 9.65% of U2OS and 78.54% of HT29 cells had positive hTERT signals. 3D animation images showed that the probe could target intranuclear hTERT in the nucleus. In mice models, fluorescence and PET imaging showed that hTERT in HT29 tumors could be efficiently visualized. In summary, we developed a new method to visualize intracellular and intranuclear proteins both in vitro and in vivo.

  5. Thymic B cells of pig fetuses and germ-free pigs spontaneously produce IgM, IgG and IgA: detection by ELISPOT method.

    PubMed Central

    Cukrowska, B; Sinkora, J; Mandel, L; Splíchal, I; Bianchi, A T; Kovárů, F; Tlaskalová-Hogenová, H

    1996-01-01

    The aim of this study was to investigate spontaneous immunoglobulin production and a pattern of isotype switching by thymic B lymphocytes (TBL) as compared with cells isolated from spleen during early ontogeny using a pig model in which B-cell development is not influenced by maternal regulatory factors. A sensitive ELISPOT assay was therefore employed to detect immunoglobulins in pig fetuses, colostrum-deprived germ-free (GF) piglets as well as conventionally (CONV) reared pigs. The first spontaneously immunoglobulin-secreting cells in the thymus were detected in 67-day-old fetuses (the length of gestation period in pigs is 114 days), their number increasing during fetal ontogeny. In contrast to fetal splenic cells, which secrete exclusively IgM, fetal thymic immunoglobulin-secreting cells were determined to undergo spontaneous isotype switching to IgG and IgA. In 28-day-old GF piglets and 3-month-old CONV pigs the number of thymic immunoglobulin-secreting cells of all isotypes was comparable to the number of thymic immunoglobulin-secreting cells detected in the newborn thymus. Considerable augmentation of IgG and IgA production by splenic immunoglobulin-secreting cells in CONV pigs was observed as compared to GF newborns and GF piglets, in which IgG- and IgA-secreting cells were detected occasionally. Our results indicate that TBL represent the first B-cell population in early fetal ontogeny spontaneously undergoing isotype switching to IgG and IgA; in the postnatal period the TBL population does not appear to be influenced by external antigenic stimuli of conventional microflora. PMID:8778038

  6. [Environmental factors in ALS].

    PubMed

    Juntas-Morales, Raul; Pageot, Nicolas; Corcia, Philippe; Camu, William

    2014-05-01

    ALS is likely to be a disorder of multifactorial origin. Among all the factors that may increase the risk of ALS, environmental ones are being studied for many years, but in the recent years, several advances have pointed to a new interest in their potential involvement in the disease process, especially for the cyanotoxin BMAA. Food containing BMAA has been found on Guam, a well-known focus of ALS/parkinsonism/dementia and high levels of BMAA have been identified into the brain of these patients. The BMAA cyanotoxin is potentially ubiquitous and have also been found into the food of patients who died from ALS both in Europe and USA. BMAA can be wrongly integrated into the protein structure during mRNA traduction, competing with serine. This may induce abnormal protein folding and a subsequent cell death. Heavy metals, such as lead or mercury may be directly toxic for neuronal cells. Several works have suggested an increased risk of ALS in individuals chronically exposed to these metals. Exposure to pesticides has been suggested to be linked to an increased risk of developing ALS. The mechanism of their toxicity is likely to be mediated by paraoxonases. These proteins are in charge of detoxifying the organism from toxins, and particularly organophosphates. To date, there are insufficient scientific data to suggest that exposure to electromagnetic fields may increase the risk of having ALS. We are particularly missing longitudinal cohorts to demonstrate that risk.

  7. Amyotrophic Lateral Sclerosis (ALS)

    MedlinePlus

    ... Pictures of ALS Mutant Proteins Support Two Major Theories About How the Disease is Caused May 2003 ... All NINDS-prepared information is in the public domain and may be freely copied. Credit to the ...

  8. Genetic Testing for ALS

    MedlinePlus

    ... Involved Donate Familial Amyotrophic Lateral Sclerosis (FALS) and Genetic Testing By Deborah Hartzfeld, MS, CGC, Certified Genetic ... guarantee a person will develop symptoms of ALS. Genetic Counseling If there is more than one person ...

  9. All About ALS

    MedlinePlus

    ... External link, please review our exit disclaimer . Subscribe All About ALS Understanding a Devastating Disorder In the ... a coffee pot, or button a shirt. Eventually, all muscles under voluntary control are affected, and people ...

  10. What Is ALS?

    MedlinePlus

    ... scarring or hardening ("sclerosis") in the region. Motor neurons reach from the brain to the spinal cord ... the body. The progressive degeneration of the motor neurons in ALS eventually leads to their demise. When ...

  11. Extensive genetics of ALS

    PubMed Central

    Calvo, Andrea; Mazzini, Letizia; Cantello, Roberto; Mora, Gabriele; Moglia, Cristina; Corrado, Lucia; D'Alfonso, Sandra; Majounie, Elisa; Renton, Alan; Pisano, Fabrizio; Ossola, Irene; Brunetti, Maura; Traynor, Bryan J.; Restagno, Gabriella

    2012-01-01

    Objective: To assess the frequency and clinical characteristics of patients with mutations of major amyotrophic lateral sclerosis (ALS) genes in a prospectively ascertained, population-based epidemiologic series of cases. Methods: The study population includes all ALS cases diagnosed in Piemonte, Italy, from January 2007 to June 2011. Mutations of SOD1, TARDBP, ANG, FUS, OPTN, and C9ORF72 have been assessed. Results: Out of the 475 patients included in the study, 51 (10.7%) carried a mutation of an ALS-related gene (C9ORF72, 32; SOD1, 10; TARDBP, 7; FUS, 1; OPTN, 1; ANG, none). A positive family history for ALS or frontotemporal dementia (FTD) was found in 46 (9.7%) patients. Thirty-one (67.4%) of the 46 familial cases and 20 (4.7%) of the 429 sporadic cases had a genetic mutation. According to logistic regression modeling, besides a positive family history for ALS or FTD, the chance to carry a genetic mutation was related to the presence of comorbid FTD (odds ratio 3.5; p = 0.001), and age at onset ≤54 years (odds ratio 1.79; p = 0.012). Conclusions: We have found that ∼11% of patients with ALS carry a genetic mutation, with C9ORF72 being the commonest genetic alteration. Comorbid FTD or a young age at onset are strong indicators of a possible genetic origin of the disease. PMID:23100398

  12. Evaluation of 12 commercial tests and the complement fixation test for Mycoplasma pneumoniae-specific immunoglobulin G (IgG) and IgM antibodies, with PCR used as the "gold standard".

    PubMed

    Beersma, Matthias F C; Dirven, Kristien; van Dam, Alje P; Templeton, Kate E; Claas, Eric C J; Goossens, Herman

    2005-05-01

    Serology and nucleic acid amplification are the main diagnostic tools for the diagnosis of Mycoplasma pneumoniae infection. Since no reference standard is generally accepted, serologic assays for M. pneumoniae have not been evaluated on a broad scale. In this study, 12 commercially available serologic assays (for immunoglobulin G [IgG] and IgM) and the complement fixation test (CFT) were evaluated by using M. pneumoniae DNA detection by real-time PCR as the "gold standard." The assays tested were Platelia EIA (Bio-Rad), SeroMP EIA (Savyon), Serion classic EIA (Virion/Serion), Biotest EIA (Biotest), Ridascreen EIA (r-Biopharm), AniLabsystems EIA (Labsystems), Novum EIA (Novum Diagnostica), Diagnosys EIA (MP products), Genzyme/Virotech EIA, ImmunoWell EIA (Genbio), ImmunoCard EIA (Meridian), and SerodiaMycoII microparticle agglutination (Fujirebio). Serum samples (n = 46) from 27 PCR-positive patients with a known first day of disease and sera (n = 33) from PCR-negative controls were obtained from prospective studies of acute lower respiratory tract infections. Additionally, control sera (n = 63) from patients with acute viral or bacterial respiratory infections other than those caused by M. pneumoniae were tested. The results showed low specificities for both the Novum and the ImmunoCard IgM assays. The IgM assays with the best performances in terms of sensitivity and specificity were AniLabsystems (77% and 92%, respectively), SeroMP (71% and 88%, respectively), and CFT (65% and 97%, respectively). Good receiver operating characteristic areas under the curve were found for CFT (0.94), the Platelia assay (0.87), and the AniLabsystems assay (0.85). We conclude that there are few commercial serologic assays for the detection of M. pneumoniae infections with appropriate performances in terms of sensitivity and specificity and that PCR has become increasingly important for the diagnosis of M. pneumoniae infections in defined groups of patients.

  13. Rapid and sensitive detection of immunoglobulin M (IgM) and IgG antibodies against canine distemper virus by a new recombinant nucleocapsid protein-based enzyme-linked immunosorbent assay.

    PubMed

    von Messling, V; Harder, T C; Moennig, V; Rautenberg, P; Nolte, I; Haas, L

    1999-04-01

    Canine distemper morbillivirus (CDV) infection causes a frequently fatal systemic disease in a broad range of carnivore species, including domestic dogs. In CDV infection, classical serology provides data of diagnostic and prognostic values (kinetics of seroconversion) and is also used to predict the optimal vaccination age of pups. Routine CDV serology is still based on time- and cost-intensive virus neutralization assays (V-NA). Here, we describe a new capture-sandwich enzyme-linked immunosorbent assay (ELISA) that uses recombinant baculovirus-expressed nucleocapsid (N) protein of a recent CDV wild-type isolate (2544/Han95) for the detection of CDV-specific antibodies in canine sera. Recombinant antigen was produced with high efficacy in Heliothis virescens larvae. The capture-sandwich ELISA enabled a clear-cut qualitative evaluation of the CDV-specific immunoglobulin G (IgG) and IgM serostatuses of 196 and 35 dog sera, respectively. Inter-rater agreement analysis (kappa = 0.988) indicated that the ELISA can be used unrestrictedly as a substitute for the V-NA for the qualitative determination of CDV-specific IgG serostatus. In an attempt to semiquantify N-specific antibodies, a one-step-dilution (alpha method) IgG-specific ELISA was implemented. Alpha values of >/=50% showed very good inter-rater agreement (kappa = 0.968) with V-NA titers of >/=1/100 50% neutralizing dose (ND50) as measured against the central European CDV wild-type isolate 2544/Han95 in canine sera originating from northern Germany. An ND50 titer of 1/100 is considered a threshold, and titers of >/=1/100 indicate a resilient, protective immunity. CDV N-specific antibodies of the IgM class were detected by the newly developed ELISA in 9 of 15 sera obtained from dogs with symptoms of acute distemper. In leucocytes of 5 of the 15 dogs (all of which were also IgM positive) CDV RNA was detected by reverse transcription (RT)-PCR. The recombinant capture-sandwich ELISA detecting N-specific antibodies

  14. Rapid and Sensitive Detection of Immunoglobulin M (IgM) and IgG Antibodies against Canine Distemper Virus by a New Recombinant Nucleocapsid Protein-Based Enzyme-Linked Immunosorbent Assay

    PubMed Central

    von Messling, Veronika; Harder, Timm C.; Moennig, Volker; Rautenberg, Peter; Nolte, Ingo; Haas, Ludwig

    1999-01-01

    Canine distemper morbillivirus (CDV) infection causes a frequently fatal systemic disease in a broad range of carnivore species, including domestic dogs. In CDV infection, classical serology provides data of diagnostic and prognostic values (kinetics of seroconversion) and is also used to predict the optimal vaccination age of pups. Routine CDV serology is still based on time- and cost-intensive virus neutralization assays (V-NA). Here, we describe a new capture-sandwich enzyme-linked immunosorbent assay (ELISA) that uses recombinant baculovirus-expressed nucleocapsid (N) protein of a recent CDV wild-type isolate (2544/Han95) for the detection of CDV-specific antibodies in canine sera. Recombinant antigen was produced with high efficacy in Heliothis virescens larvae. The capture-sandwich ELISA enabled a clear-cut qualitative evaluation of the CDV-specific immunoglobulin G (IgG) and IgM serostatuses of 196 and 35 dog sera, respectively. Inter-rater agreement analysis (κ = 0.988) indicated that the ELISA can be used unrestrictedly as a substitute for the V-NA for the qualitative determination of CDV-specific IgG serostatus. In an attempt to semiquantify N-specific antibodies, a one-step-dilution (alpha method) IgG-specific ELISA was implemented. Alpha values of ≥50% showed very good inter-rater agreement (κ = 0.968) with V-NA titers of ≥1/100 50% neutralizing dose (ND50) as measured against the central European CDV wild-type isolate 2544/Han95 in canine sera originating from northern Germany. An ND50 titer of 1/100 is considered a threshold, and titers of ≥1/100 indicate a resilient, protective immunity. CDV N-specific antibodies of the IgM class were detected by the newly developed ELISA in 9 of 15 sera obtained from dogs with symptoms of acute distemper. In leucocytes of 5 of the 15 dogs (all of which were also IgM positive) CDV RNA was detected by reverse transcription (RT)-PCR. The recombinant capture-sandwich ELISA detecting N-specific antibodies of the

  15. ALS superbend magnet system

    SciTech Connect

    Zbasnik, J.; Wang, S.T.; Chen, J.Y.; DeVries, G.J.; DeMarco, R.; Fahmie, M.; Geyer, A.; Green, M.A.; Harkins, J.; Henderson, T.; Hinkson, J.; Hoyer, E.H.; Krupnick, J.; Marks, S.; Ottens, F.; Paterson, J.A.; Pipersky, P.; Portmann, G.; Robin, D.A.; Schlueter, R.D.; Steier, C.; Taylor, C.E.; Wahrer, R.

    2000-09-15

    The Lawrence Berkeley National Laboratory is preparing to upgrade the Advanced Light Source (ALS) with three superconducting dipoles (Superbends). In this paper we present the final magnet system design which incorporates R&D test results and addresses the ALS operational concerns of alignment, availability, and economy. The design incorporates conduction-cooled Nb-Ti windings and HTS current leads, epoxy-glass suspension straps, and a Gifford-McMahon cryocooler to supply steady state refrigeration. We also present the current status of fabrication and testing.

  16. Modulus measurements in ordered Co-Al, Fe-Al, and Ni-Al alloys

    NASA Technical Reports Server (NTRS)

    Harmouche, M. R.; Wolfenden, A.

    1985-01-01

    The composition and/or temperature dependence of the dynamic Young's modulus for the ordered B2 Co-Al, Fe-Al, and Ni-Al aluminides has been investigated using the piezoelectric ultrasonic composite oscillator technique (PUCOT). The modulus has been measured in the composition interval 48.49 to 52.58 at. pct Co, 50.87 to 60.2 at. pct Fe, and 49.22 to 55.95 at. pct Ni for Co-Al, Fe-Al, and Ni-Al, respectively. The measured values for Co-Al are in the temperature interval 300 to 1300 K, while those for the other systems are for ambient temperature only. The data points show that Co-Al is stiffer than Fe-Al, which is stiffer than Ni-Al. The data points for Fe-Al and Ni-Al are slightly higher than those reported in the literature.

  17. Al Shanker Remembers.

    ERIC Educational Resources Information Center

    American Educator, 2000

    2000-01-01

    In a 1996 interview shortly before his death, Al Shanker, longtime president of the American Federation of Teachers, discussed such topics as: his own educational experiences; how he learned about political fighting in the Boy Scouts; the appeal of socialism; multinational corporations and the nation state; teaching tough students; and John Dewey…

  18. Fosetyl-al

    Integrated Risk Information System (IRIS)

    Fosetyl - al ; CASRN 39148 - 24 - 8 Human health assessment information on a chemical substance is included in the IRIS database only after a comprehensive review of toxicity data , as outlined in the IRIS assessment development process . Sections I ( Health Hazard Assessments for Noncarcinogenic Ef

  19. ALS renewal moves forward

    NASA Astrophysics Data System (ADS)

    Falcone, R. W.; Feinberg, B.; Hussain, Z.; Kirz, J.; Krebs, G. F.; Padmore, H. A.; Robin, D. S.; Robinson, A. L.

    2007-11-01

    As the result of an extensive long-term planning process involving all its stakeholders—management, staff, and users—the ALS has seen its future and is aggressively moving ahead to implement its vision for keeping the facility at the cutting edge for the next 2-3 decades. The evolving strategic plan now in place aims to renew the ALS so it can address a new generation of fundamental questions about size dependent and dimensional-confinement phenomena at the nanoscale; correlation and complexity in physical, biological, and environmental systems; and temporal evolution, assembly, dynamics and ultrafast phenomena. The renewal spans three areas: (1) increased staffing at beamlines to support the growing user community and safety professionals to keep an increasingly complex facility hazard free; (2) implementing advances in accelerator, insertion device, beamline, and detector technology that will make it possible for ALS users to address emerging grand scientific and technological challenges with incisive world-class tools; and (3) construction of a user support building and guest housing that will increase the safety and user friendliness of the ALS by providing users office, meeting, experiment staging, and laboratory space for their work and on-site accommodations at reasonable rates.

  20. Serum levels of immunoglobulins (IgG, IgA, IgM) in a general adult population and their relationship with alcohol consumption, smoking and common metabolic abnormalities

    PubMed Central

    Gonzalez-Quintela, A; Alende, R; Gude, F; Campos, J; Rey, J; Meijide, L M; Fernandez-Merino, C; Vidal, C

    2008-01-01

    The present study investigated serum immunoglobulin (Ig) concentrations in relation to demographic factors, common habits (alcohol consumption and smoking) and metabolic abnormalities in an adult population-based survey including 460 individuals. Serum levels of interleukin (IL)-6, a marker of inflammation, were also determined. After adjusting for confounders, male sex was associated positively with IgA levels and negatively with IgM levels. Age was associated positively with IgA and IgG levels. Smoking was associated negatively with IgG levels. Heavy drinking was associated positively with IgA levels. Metabolic abnormalities (obesity and metabolic syndrome) were associated positively with IgA levels. Abdominal obesity and hypertriglyceridaemia were the components of metabolic syndrome associated most strongly with serum IgA. Heavy drinkers with metabolic syndrome showed particularly high serum IgA levels. Serum IL-6 levels were correlated positively with IgA and IgG concentrations. It is concluded that sex, age, alcohol consumption, smoking and common metabolic abnormalities should be taken into account when interpreting serum levels of IgA, IgG and IgM. PMID:18005364

  1. Biodistribution and tumor localization of 111In-labeled unmodified and modified F(ab')2 fragments of human monoclonal IgM (16.88) in a nude mouse model.

    PubMed

    Quadri, S M; Siddiqui, A; Klein, J L; Vriesendorp, H M

    1995-05-01

    Unmodified F(ab')2 and modified Fab'-BMH-Fab' fragments of human monoclonal IgM (16.88) were compared for biodistribution and tumor localization in nude mice bearing LS-174T human colon carcinoma xenografts. Although both unmodified and modified fragments of IgM cleared rapidly from the blood, the radioactivity retentions for each fragment in liver and kidney were significantly different. Kidney uptake of the modified fragment was about 4-fold lower than kidney uptake of the unmodified fragment. Radioactivity uptake in liver was 2-4-fold higher for the modified fragment. Lower liver and higher kidney uptake of unmodified fragments reflected the labile disulfide linkage of F(ab')2 in their hinge region and the subsequent behavior of the Fab' fragments resulting from the reduction of the disulfide linkage. Higher liver and lower kidney retention of modified fragments, on the other hand, resulted from the different cleavage mechanism of the stable thioether linkage. Tumor targeting was similar for unmodified and modified fragments at approx. 4% of injected dose per gram. These results indicate that the changes in fragment linkage chemistry may provide different pharmacokinetic patterns in vivo and improve the therapeutic application of radiolabeled fragments in human patients.

  2. Enzyme linked immunosorbent assay (ELISA) for the detection of IgG, IgM, IgE and IgA against Cysticercus cellulosae in cerebrospinal fluid of patients with neurocysticercosis.

    PubMed

    Odashima, Newton Satoru; Takayanagui, Osvaldo Massaiti; Figueiredo, José Fernando de Castro

    2002-06-01

    The objective of this study was to analyze different immunoglobulins classes (IgG, IgM, IgE and IgA) against Cysticercus cellulosae in the cerebrospinal fluid (CSF), through enzyme linked immunosorbent assay (ELISA), correlating them to clinical and tomographic profiles in patients with neurocysticercosis (NCC). Eighty-five specimens of CSF were obtained from 43 cases with NCC (26 with the active form and 17 with the inactive form) and from 42 patients with other neurological diseases. The inactive form of NCC presented a profile in CSF similar to the group without NCC. The active form of NCC presented elevation of specific immunoglobulins (IgG, IgM, IgE, and IgA) in decreasing order, with the highest values being detected among the cases with intraventricular cysts, or with inflammation signs in CSF or in those with multiple clinical manifestations. The highest sensitivity and specificity were obtained with ELISA-IgG (88.5% and 93.2%, respectively). This study confirmed the importance of ELISA in the immunologic diagnosis of NCC.

  3. Ausbildung als zentrale Aufgabe

    NASA Astrophysics Data System (ADS)

    Krämer, Walter; Schmerbach, Sibylle

    Anders als Lesen und Schreiben zählen Grundkenntnisse in Statistik heute noch nicht zu den Voraussetzungen einer sinnvollen Teilhabe am Sozialgeschehen. Und auch in der akademischen Statistik-Ausbildung gibt es noch einiges zu tun. Das vorliegende Kapitel zeichnet die Geschichte dieser akademischen Ausbildung an deutschen Universitäten nach dem zweiten Weltkrieg nach, stellt aktuelle Defizite vor und weist auf mögliche Verbesserungen hin.

  4. Clonal analysis of a human antibody response. Quantitation of precursors of antibody-producing cells and generation and characterization of monoclonal IgM, IgG, and IgA to rabies virus

    PubMed Central

    1990-01-01

    We quantitated and characterized the changes in the human B cell repertoire, at the clonal level, before and after immunization with rabies virus. Moreover, we generated 10 monoclonal cell lines producing IgM, IgG, and IgA antibodies to the virus. We found that in healthy subjects, not previously exposed to the virus, nearly 2% of the circulating B lymphocytes were committed to the production of antibodies that bound the virus. These B cells expressed the surface CD5 molecule. The antibodies they produced were polyreactive IgM that displayed a relatively low affinity for the virus components (Kd, 1.0- 2.4 x 10(-6) g/microliters). After immunization, different anti-virus (IgG and IgA) antibody-producing cells consistently appeared in the circulation and increased from less than 0.005% to greater than 10% of the total B cells committed to the production of IgG and IgA, respectively. Most of such B cells do not express CD5 and produce monoreactive antibodies of high affinity for rabies virus (Kd, 6.5 x 10(-9) to 1.2 x 10(-10) g/microliters). One of these IgG mAbs efficiently neutralized rabies virus in vitro and in vivo, as detailed elsewhere (Dietzschold, B., P. Casali, Y. Ueki, M. Gore, C. E. Rupprecht, A. L. Notkins, and H. Koprowski, manuscript submitted for publication). Hybridization experiments using probes specific for the different human V gene segment families revealed that cell precursors producing low affinity IgM binding to rabies virus utilized a restricted number of VH gene segments (i.e., only members of the VHIIIb subfamily), whereas cell precursors producing high affinity IgG and IgA to rabies virus utilized an assortment of different VH gene segments (i.e., members of the VHI, VHIII, VHIV, and VHVI families and VHIIIb subfamily). In conclusion, our studies show that EBV transformation in conjunction with limiting dilution technology and somatic cell hybridization techniques are useful methods for quantitating, at the B cell clonal level, the human

  5. AL Amyloidosis and Agent Orange

    MedlinePlus

    ... for survivors' benefits . Research on AL amyloidosis and herbicides The Health and Medicine Division (formally known as ... to the compounds of interest found in the herbicide Agent Orange and AL amyloidosis." VA made a ...

  6. Modeling neuronal vulnerability in ALS.

    PubMed

    Roselli, Francesco; Caroni, Pico

    2014-08-20

    Using computational models of motor neuron ion fluxes, firing properties, and energy requirements, Le Masson et al. (2014) reveal how local imbalances in energy homeostasis may self-amplify and contribute to neurodegeneration in ALS.

  7. Al Jazirah, Sudan

    NASA Technical Reports Server (NTRS)

    2008-01-01

    Al Jazirah (also Gezira) is one of the 26 states of Sudan. The state lies between the Blue Nile and the White Nile in the east-central region of the country. It is a well populated area suitable for agriculture. The area was at the southern end of Nubia and little is known about its ancient history and only limited archaeological work has been conducted in this area. The region has benefited from the Gezira Scheme, a program to foster cotton farming begun in 1925. At that time the Sennar Dam and numerous irrigation canals were built. Al Jazirah became the Sudan's major agricultural region with more than 2.5 million acres (10,000 km) under cultivation. The initial development project was semi-private, but the government nationalized it in 1950. Cotton production increased in the 1970s but by the 1990s increased wheat production has supplanted a third of the land formerly seeded with cotton.

    The image was acquired December 25, 2006, covers an area of 56 x 36.4 km, and is located near 14.5 degrees north latitude, 33.1 degrees east longitude.

    The U.S. science team is located at NASA's Jet Propulsion Laboratory, Pasadena, Calif. The Terra mission is part of NASA's Science Mission Directorate.

  8. Accuracy of IgM antibody testing, FQ-PCR and culture in laboratory diagnosis of acute infection by Mycoplasma pneumoniae in adults and adolescents with community-acquired pneumonia

    PubMed Central

    2013-01-01

    Background Diagnosis of community-acquired pneumonia (CAP) caused by Mycoplasma pneumoniae in adults and adolescents is hampered by a lack of rapid and standardized tests for detection. Methods CAP patients from 12 teaching hospitals were prospectively and consecutively recruited. Basic and clinical information, throat swabs and paired sera were collected. Mycoplasma pneumoniae was detected by IgG and IgM antibody tests, fluorescence quantitative polymerase chain reaction (FQ-PCR) and culture. A comparative study of the diagnostic values of three methods, including sensitivity, specificity, positive and negative predictive values and positive likelihood ratio (PLR) was conducted. A fourfold or greater increase of IgG antibody titers of paired sera was set as the diagnostic “gold standard”. Results One hundred and twenty-five CAP patients (52.8% males, median age 47 years, range 14–85) were enrolled. Twenty-seven (21.6%) patients were diagnosed with acute Mycoplasma pneumoniae infections by the “gold standard”. Specificity values of all three methods were around 90%. An increasing trend of sensitivity, positive predictive value and PLR was found, with the lowest in IgM testing (7.4%, 28.6% and 1.45), intermediate in FQ-PCR (40.7%, 50% and 3.63), and highest in culture (55.6%, 75% and 10.9). Conclusions In the defined group of patients, there was a good agreement between positive rate of MP cultivation of throat swabs and acute M. pneumoniae infection (PLR of 10.9). Since the sensitivity is low in all of the evaluated methods, the logical approach would be to incorporate PCR, culture and serological tests for optimum diagnosis of acute Mycoplasma pneumoniae infections in adults and adolescents. PMID:23578215

  9. Identification and real-time expression analysis of selected Toxoplasma gondii in-vivo induced antigens recognized by IgG and IgM in sera of acute toxoplasmosis patients

    PubMed Central

    2013-01-01

    Background Toxoplasma gondii is an obligate intracellular zoonotic parasite of the phylum Apicomplexa which infects a wide range of warm-blooded animals, including humans. In this study in-vivo induced antigens of this parasite was investigated using in-vivo induced antigen technology (IVIAT) and pooled sera from patients with serological evidence of acute infection. Methods The pooled sera was first pre-absorbed against three different preparations of antigens from in-vitro-grown cells of each T. gondii and E. coli XL1-Blue MRF’, subsequently it was used to screen T. gondii cDNA phage expression library. Positive clones from each group were subjected to quantitative real-time PCR expression analysis on mRNA of in-vivo and in-vitro grown parasites. Results A total of 29 reactive clones from each IgM and IgG immunoscreenings were found to have high homology to T. gondii genes. Quantitative real-time PCR expression analysis showed that 20 IgM-detected genes and 11 IgG-detected genes were up-regulated in-vivo relative to their expression levels in-vitro. These included genes encoding micronemes, sterol-regulatory element binding protein site, SRS34A, MIC2-associated protein M2AP, nucleoredoxin, protein phosphatase 2C and several hypothetical proteins. A hypothetical protein (GenBank accession no. 7899266) detected by IgG had the highest in-vivo over in-vitro fold change of 499.86; while another up-regulated hypothetical protein (GenBank accession no. 7898829) recognized by IgM showed high sensitivity (90%) and moderate specificity (70%) in detecting T. gondii antibodies when tested with 20 individual serum samples. Conclusion The highly up-regulated genes and the corresponding proteins, in particular the hypothetical proteins, may be useful in further studies on understanding the disease pathogenesis and as potential vaccine candidates. PMID:23800344

  10. [BPO-Specific, complement-dependant cell-lysis of differently sensitized sheep red cells: evaluation of haptenic groups and their influence on IgM and IgG-induced lysis (author's transl)].

    PubMed

    Wiedermann, G; Stemberger, H; Förster, O; Müller, M

    1976-04-01

    Sheep erythrocytes were coated with bencylpenicilloyl-(BPO)groups. Different incubation periods resulted in erythrocyte preparations with different hapten density. Complement dependent lysis induced by IgM or IgG antibodies was studied with the cell preparations. The calculation of hapten density on the erythrocyte surface was not possible by direct measurement of coupled radioactive BPO since more than 90% of radioactive material was found in the soluble supernatant after osmotic cell lysis and less than 10% was fixed to the cellular membrane. Measurement of membrane bound immunologically relevant BPO-groups was achieved, therefore, by comparison of the inhibitory capacity of the test cells with that of a standard cell preparation. The latter consisted of tannic acid treated erythrocytes coated with protein complexed radioactive BPO. Surface hapten density of the different target cell preparations varied between 1.9 x 10(5) and 4.8 10(5) BPO-groups per cell depending on the time of incubation. Complement dependent antibody mediated cell lysis was significantly reduced by reduction of haptenic sites per target cell, IgG induced lysis being much more affected than hemolysis induced by IgM antibodies. Statistical calculations led to the conclusion that 18,000 protein islets per cell bearing 4 or more BPO-groups are not sufficient for hemolysis induced by IgG antibodies. 48,000 protein islets with this hapten density are necessary for "optimal" sensitization. IgG antibodies must be apparently bound to the cell surface in bivalent form.

  11. Development of a Highly Specific IgM Enzyme-Linked Immunosorbent Assay for Bartonella henselae Using Refined N-Lauroyl-Sarcosine-Insoluble Proteins for Serodiagnosis of Cat Scratch Disease.

    PubMed

    Otsuyama, Ken-Ichiro; Tsuneoka, Hidehiro; Kondou, Kaori; Yanagihara, Masashi; Tokuda, Nobuko; Shirasawa, Bungo; Ichihara, Kiyoshi

    2016-04-01

    The conventional anti-Bartonella henselaeIgM enzyme-linked immunosorbent assay (IgM-ELISA) methods for diagnosing cat scratch disease (CSD) remain poor in both sensitivity and specificity. We sought to develop an IgM-ELISA with improved accuracy in the serodiagnosis of CSD by exploring the antigens that are most suitable for an ELISA. We prepared 5 different protein antigens: antigen I (sonicatedB. henselaewhole-cell antigen), antigen II (N-lauroyl-sarcosine-insoluble antigen), antigen III (processed sarcosine-soluble antigen), and antigen IV and antigen V (sarcosine-insoluble and sarcosine-soluble antigens refined by DEAE-Sepharose Fast Flow ion-exchange chromatography). The IgM antibodies in the sera of 47 patients with clinically suspected CSD (24 definite, 23 suspected) and of 85 healthy individuals were examined by ELISAs using the 5 antigens, and the results were compared with those of an IgM indirect fluorescent antibody assay (IgM-IFA). In a reference panel, which consisted of 5 positive and 5 negative sera, antigen I and antigen III failed to distinguish between the two statuses, whereas the other three antigens succeeded in distinguishing between them. When the cutoff value was set at the 98th percentile of the ELISA index for healthy individuals, the sensitivity of IgM-IFA for the 24 cases of definite CSD was 54%, whereas the sensitivities of the IgM-ELISAs with antigen II, IV, and V were 75%, 83%, and 75%, respectively. The sensitivities of these three IgM-ELISAs for all 47 of the clinically suspected cases were 49%, 64%, and 51%, respectively. In contrast, the sensitivity of IgM-IFA was 28%. These results indicate that the refined sarcosine-insoluble proteins (antigen IV), which possessed the highest specificity among the 5 antigens, are the most appropriate for developing an IgM-ELISA for the highly specific serodiagnosis of CSD.

  12. Decreased IgA+ B cells population and IgA, IgG, IgM contents of the cecal tonsil induced by dietary high fluorine in broilers.

    PubMed

    Liu, Juan; Cui, Hengmin; Peng, Xi; Fang, Jing; Zuo, Zhicai; Deng, Junliang; Wang, Hesong; Wu, Bangyuan; Deng, Yuanxin; Wang, Kangping

    2013-05-02

    Fluoride is an environmental and industrial pollutant that affects various organs in humans and animals. The cecal tonsil is an important component of the mucosal immune system and performs important and unique immune functions. In the present study, we investigated the effects of dietary high fluorine on the quantities of IgA+ B cells in the cecal tonsil by immunohistochemistry, and the immunoglobulin A (IgA), immunoglobulin G (IgG) and immunoglobulin M (IgM) contents in the cecal tonsil by ELISA. A total of 280 one-day-old avian broilers were divided into four groups and fed on a corn-soybean basal diet as control diet (fluorine 22.6 mg/kg) or the same diet supplemented with 400, 800 and 1,200 mg/kg fluorine (high fluorine groups I, II and III) in the form of sodium fluoride, respectively, throughout a 42-day experimental period. The results showed that the quantities of IgA+ B cells were lower (p < 0.05 or p < 0.01) and the IgA, IgG, and IgM contents were decreased (p < 0.05 or p < 0.01) in high fluorine groups II and III in comparison with those of control group. It was concluded that dietary fluorine, in the 800-1,200 mg/kg range, could reduce the numbers of the IgA+ B cells and immunoglobulin contents in the cecal tonsil, implying the local mucosal immune function was ultimately impacted in broilers.

  13. Effects of heat stress on peripheral T and B lymphocyte profiles and IgG and IgM serum levels in broiler chickens vaccinated for Newcastle disease virus.

    PubMed

    Honda, Bruno Takashi Bueno; Calefi, Atilio Sersun; Costola-de-Souza, Carolina; Quinteiro-Filho, Wanderley Moreno; da Silva Fonseca, Juliana Garcia; de Paula, Viviane Ferraz; Palermo-Neto, João

    2015-10-01

    Multiple factors, such as environment, nutritional status, and disease, induce stress in animals during livestock production. It has been shown that poultry exposed to stressors for prolonged periods had decreases in their performance parameters, mortality and decreased host resistance to pathogenic agents. It seems that early age stress may have long-lasting impact and could possibly modify the expression of their genetic potential on growth performance and immunity. This study aimed to discuss the effects of early-age heat stress on the blood lymphocyte phenotypes (B and T lymphocytes) and plasma immunoglobulin levels (IgM and IgG) in chickens vaccinated against paramixovirus of the Newcastle (NC) disease (LaSota strain). For this purpose, 96 male chickens (Cobb) were divided into 4 groups: 1) control (C), 2) heat-stressed (HS), 3) control vaccinated (C/V), and 4) heat-stressed and Vaccinated (HS/V). The NC vaccine was administered twice on experimental day (ED) 7 and ED14, and the heat stress (38 ± 1°C) was applied from ED2 to ED6. The data showed that HS increased the corticosterone serum levels in the HS group compared with the control groups (C and C/V groups). At ED7, increased concentrations of IgM were observed in birds in the HS and HS/V groups compared with C and C/V animals; chickens from the HS/V group presented increased IgG levels compared with those in the birds of the C group. The heat stress shifted the immune cell profile from B-lymphocyte to a T-cytotoxic and T-helper lymphocyte profile, and this immune cell pattern persisted until the end of the study period. It was concluded that heat stress immunomodulated the immune function response of the chickens to the NC disease vaccine challenge. PMID:26362974

  14. ALS Performance Summary - Update

    SciTech Connect

    Waters, A M; Brown, W D; Martz, Jr., H E

    2004-09-30

    High Energy Density Physics (HEDP) experiments play an important role in corroborating the improved physics codes that underlie LLNL's Stockpile Stewardship mission. Conducting these experiments, whether on the National Ignition Facility (NIF) or another national facility such as Omega, will require not only improvement in the diagnostics for measuring the experiment, but also detailed knowledge of the as-built target components and assemblies themselves. To assist in this effort, a defined set of well-known reference standards designed to represent a range of HEDP targets have been built and are being used to quantify the performance of different characterization techniques [Hibbard, et al. 2004]. Without the critical step of using reference standards for qualifying characterization tools there can be no verification of either commercial or internally-developed characterization techniques and thus an uncertainty in the input to the physics code models would exist.

  15. Theoretical studies of AlF, AlCl, and AlBr

    NASA Technical Reports Server (NTRS)

    Langhoff, Stephen R.; Bauschlicher, Charles W., Jr.; Taylor, Peter R.

    1988-01-01

    Spectroscopic constants have been obtained for the lowest six singlet and lowest five triplet states of AlF and AlCl. The results suggest that the correct ordering of the triplet manifold in these molecules is: a 3Pi, b 3Sigma(+), c 3Sigma(+), d 3Pi, and e 3Delta. Radiative lifetimes have been determined for the excited states, and the A 1Pi to X 1Sigma(+) transition in AlF, AlCl, and AlBr has been examined in detail. A-X transition moment functions, Einstein coefficients, and A 1Pi vibrational lifetimes have been obtained for AlF and AlCl.

  16. The effects of ageing on the immune response to Schistosoma haematobium and hookworm by measuring circulating immune complexes, C3, IgG, IgA and IgM levels in residents of Omi dam area of Kogi State, Nigeria.

    PubMed

    Oyeyinka, G O; Awogun, I A; Akande, T M; Awarun, J A; Arinola, O G; Salimonu, L S

    2003-09-01

    In this study, the effects of infestation (with Schistosoma haematobium or hookworm) during host ageing on the serum levels of circulating immune complexes (CIC), C3, IgG, IgA and IgM were examined in residents of Omi dam area of Kogi state, Nigeria. S. haematobium-infested and hookworm-infested individuals showed no significant alteration in the levels of CIC, C3, IgG, IgA and IgM in comparison with controls. These levels were the same in infested subjects and controls even when the patients were pooled. Infested old people had the same concentrations of serum CIC, C3 IgG and IgM in comparison with infested young people but IgA levels were higher in the aged group (t=2.100; P<0.05); and were significantly correlated with age (r=0.301; P<0.05). No significant increase in CIC levels with rising age (r=0.123; P>0.20) was observed in the overall population of infested subjects; and infestation in old age did not alter CIC, C3, IgG, IgA and IgM levels in comparison with uninfested young people. For the uninfested, IgG, IgA and IgM values were similar in the aged and the young but the levels of CIC were higher (t=2.156; P<0.05; r=0.280; P<0.05) and C3 lower (t=3.313; P<0.01; r=-0.236; P>0.10) in the aged. The results of this study suggest that the elevated CIC levels found in old people is age-related; and that the contribution of parasitic infestation to these raised levels is uncertain.

  17. Alexandria (Al Iskandariya), Egypt

    NASA Technical Reports Server (NTRS)

    2002-01-01

    This image of Alexandria was taken by astronauts on board the International Space Station in December 2000 using an Electronic Still Camera. A wider-angle view (STS088-739-90) taken from the Space Shuttle in December 1998 is available for context. Alexandria (Al Iskandariya) occupies a T-shaped peninsula and strip of land separating the Mediterranean from Lake Mariout. Originally the town was built upon a mole (stone breakwater) called Heptastadium, which joined the island of Pharos (see referenced website, below) to the mainland. Since then sedimentary deposits have widened the mole. Since 1905, when the 370,000 Alexandrians lived in an area of about 4 sq km between the two harbors, the city (population 4 million; see referenced website, below) has grown beyond its medieval walls and now occupies an area of about 300 sq km. The Mahmudiya Canal, connecting Alexandria with the Nile, runs to the south of the city and, by a series of locks, enters the harbor of the principal port of Egypt (note ships). The reddish and ochre polygons west of Lake Mariout are salt-evaporation, chemical-storage, and water-treatment ponds within the coastal lagoon. Reference Youssef Halim and Fatma Abou Shouk, 2000, Human impacts on Alexandria's marine environment: UNESCO, Coastal Regions and Small Islands Unit (CSI), Coastal Management Sourcebooks 2 (accessed December 20, 2000) Additional photographs taken by astronauts can be viewed at NASA-JSC's Gateway to Astronaut Photography of Earth. Image ISS001-ESC-5025 provided by the Earth Sciences and Image Analysis Laboratory, Johnson Space Center.

  18. Thermal Modeling of Al-Al and Al-Steel Friction Stir Spot Welding

    NASA Astrophysics Data System (ADS)

    Jedrasiak, P.; Shercliff, H. R.; Reilly, A.; McShane, G. J.; Chen, Y. C.; Wang, L.; Robson, J.; Prangnell, P.

    2016-07-01

    This paper presents a finite element thermal model for similar and dissimilar alloy friction stir spot welding (FSSW). The model is calibrated and validated using instrumented lap joints in Al-Al and Al-Fe automotive sheet alloys. The model successfully predicts the thermal histories for a range of process conditions. The resulting temperature histories are used to predict the growth of intermetallic phases at the interface in Al-Fe welds. Temperature predictions were used to study the evolution of hardness of a precipitation-hardened aluminum alloy during post-weld aging after FSSW.

  19. Thermal Modeling of Al-Al and Al-Steel Friction Stir Spot Welding

    NASA Astrophysics Data System (ADS)

    Jedrasiak, P.; Shercliff, H. R.; Reilly, A.; McShane, G. J.; Chen, Y. C.; Wang, L.; Robson, J.; Prangnell, P.

    2016-09-01

    This paper presents a finite element thermal model for similar and dissimilar alloy friction stir spot welding (FSSW). The model is calibrated and validated using instrumented lap joints in Al-Al and Al-Fe automotive sheet alloys. The model successfully predicts the thermal histories for a range of process conditions. The resulting temperature histories are used to predict the growth of intermetallic phases at the interface in Al-Fe welds. Temperature predictions were used to study the evolution of hardness of a precipitation-hardened aluminum alloy during post-weld aging after FSSW.

  20. Philosophieren als Unterrichtsprinzip im Mathematikunterricht

    NASA Astrophysics Data System (ADS)

    Meerwaldt, Diana

    Philosophieren und Mathematik scheinen zunächst gegensätzliche Bereiche zu sein, die sich kaum vereinbaren lassen. Dies trifft für eine Auffassung zu, die Philosophieren als "Gerede" disqualifiziert und Mathematik als eine reine "Formelwissenschaft" begreift. Beide Auffassungen werden den Gegenständen nicht gerecht.

  1. The Advanced Launch System (ALS)

    NASA Astrophysics Data System (ADS)

    Eldred, Charles H.

    The Advanced Launch System (ALS) is an unmanned vehicle that will achieve low hardware cost by using a reusable booster stage which flies back to the launch site, and a core stage in which the rocket engines and redundant avionics are in a module that is returned to earth and recovered for reuse. The booster's utilization of liquid propellant instead of solid propellant will help lower the consumable costs. The ALS also includes launch processing and flight control facilities, necessary support equipment, and ground- and flight-operations infrastructure. The ALS program studies show that, through the ALS, the United States can launch a major Mars initiative economically and with confidence. It is estimated that the objective ALS can be operational in the late 1990s.

  2. Quantum dots formed in InSb/AlAs and AlSb/AlAs heterostructures

    NASA Astrophysics Data System (ADS)

    Abramkin, D. S.; Rumynin, K. M.; Bakarov, A. K.; Kolotovkina, D. A.; Gutakovskii, A. K.; Shamirzaev, T. S.

    2016-06-01

    The crystal structure of new self-assembled InSb/AlAs and AlSb/AlAs quantum dots grown by molecularbeam epitaxy has been investigated by transmission electron microscopy. The theoretical calculations of the energy spectrum of the quantum dots have been supplemented by the experimental data on the steady-state and time-resolved photoluminescence spectroscopy. Deposition of 1.5 ML of InSb or AlSb on the AlAs surface carried out in the regime of atomic-layer epitaxy leads to the formation of pseudomorphically strained quantum dots composed of InAlSbAs and AlSbAs alloys, respectively. The quantum dots can have the type-I and type-II energy spectra depending on the composition of the alloy. The ground hole state in the quantum dot belongs to the heavy-hole band and the localization energy of holes is much higher than that of electrons. The ground electron state in the type-I quantum dots belongs to the indirect X XY valley of the conduction band of the alloy. The ground electron state in the type-II quantum dots belongs to the indirect X valley of the conduction band of the AlAs matrix.

  3. Association of serum IgM kappa monoclonicity in patients with Sjögren's syndrome with an increased proportion of kappa positive plasma cells infiltrating the labial minor salivary glands.

    PubMed Central

    Moutsopoulos, H M; Tzioufas, A G; Bai, M K; Papadopoulos, N M; Papadimitriou, C S

    1990-01-01

    Minor salivary gland biopsy specimens from 11 patients with primary Sjögren's syndrome with circulating monoclonal IgM kappa cryoglobulins, seven without cryoglobulins, and four patients with rheumatoid arthritis and Sjögren's syndrome (one with monoclonal and three with polyclonal cryoglobulins) were examined by the peroxidase antiperoxidase bridge technique, using antihuman kappa and lambda antibodies. In 6/11 patients with primary Sjögren's syndrome and in one patient with Sjögren's syndrome and rheumatoid arthritis with monoclonal cryoglobulins a predominance of plasma cells containing intracytoplasmic kappa light chains was found (kappa:lambda greater than 3:1). Two of those seven patients had immunohistological features of immunocytomas. In the other five patients with circulating monoclonal cryoglobulins the kappa:lambda ratio of positive cells did not exceed 3:1, while six out of seven patients without cryoglobulins and the patients with rheumatoid arthritis and Sjögren's syndrome with polyclonal cryoglobulins had almost equal numbers of kappa and lambda stained cells. One of seven patients with primary Sjögren's syndrome without cryoglobulins had an increased number of lambda light chain positive cells, indicating a non-secretory lambda monoclonal population. These findings suggest that the main area of B cell monoclonal expansion in primary Sjögren's syndrome may be the affected exocrine glands. PMID:2124096

  4. Development of a novel protein biochip enabling validation of immunological assays and detection of serum IgG and IgM antibodies against Treponema pallidum pathogens in the patients with syphilis.

    PubMed

    Huang, Na-Li; Ye, Lei; Schneider, Marion E; Du, Yi-Xin; Xu, Yuan-Hong; Fan, Li-Bin; Du, Wei-Dong

    2016-01-15

    In this study, we developed a novel protein biochip methodology that was characterized by dithiobis (succinimidyl undecanoate) (DSU) and specialized for detection of serum IgG and IgM antibodies against Treponema pallidum pathogens in the patients with syphilis, respectively. The biochips were validated by a dimension of atomic force microscope (AFM). The visualized detection limit of IgG antibody on the biochip was 0.39μg/ml. Finally, 286 serum samples from the patients with syphilis were simultaneously tested on the rTpN15-17-47 coated biochips. The results were evaluated in comparison with the assays of T. pallidum particle agglutination (TPPA) and the toluidine red unheated serum test (TRUST). The result demonstrated that the relative positive rate in the 286 patients by biochip was 99.0%, similar to that by TPPA (97.9%, P>0.05) and higher than that by TRUST, (76.2%, P<0.01). The detection specificities were 100% for the biochip and the TPPA and 97.0% for the TRUST. Thus, the protein biochip would provide a useful platform not only for enabling concurrent detection of the infectious antibodies directed against T. pallidum on a larger scale, but also for monitoring therapy modality of the disease.

  5. Medical application of 26Al

    NASA Astrophysics Data System (ADS)

    Steinhausen, C.; Gerisch, P.; Heisinger, B.; Hohl, Ch.; Kislinger, G.; Korschinek, G.; Niedermayer, M.; Nolte, E.; Dumitru, M.; Alvarez-Brückmann, M.; Schneider, M.; Ittel, T. H.

    1996-06-01

    Accelerator mass spectrometry (AMS) measurements with 26Al as tracer were performed in order to study the aluminium metabolism and anomalies in the human body and in rats. In particular, the differences between healthy volunteers and patients with renal failure were investigated. The obtained data points of 26Al in blood and urine were described by an open compartment model with three peripheral compartments. It was found that the minimum of peripheral compartments needed to describe 26Al concentrations in blood and urine over a time period of three years is at least three.

  6. In memory of Al Cameron

    NASA Astrophysics Data System (ADS)

    Cowan, John; Truran, James W.

    Al Cameron, who died recently (October 3, 2005) at 80, was one of the giants in astrophysics. His insights were profound and his interests were wide-ranging. Originally trained as a nuclear physicist, he made major contributions in a number of fields, including nuclear reactions in stars, nucleosynthesis, the abundances of the elements in the Solar System, and the origin of the Solar System and the Moon. In 1957, Cameron and, independently, Burbidge, Burbidge, Fowler and Hoyle, wrote seminal papers on nuclear astrophysics. Most of our current ideas concerning ele- ment formation in stars have followed from those two pioneering and historical works. Al also made many contributions in the field of Solar System physics. Particularly noteworthy in this regard was Cameron's work on the formation of the Moon. Al was also a good friend and mentor of young people. Al Cameron will be missed by many in the community both for his scientific contributions and for his friendship.

  7. Determination by enzyme-linked immunosorbent assay of immunoglobulin G (IgG), IgM, and IgA to Brucella melitensis major outer membrane proteins and whole-cell heat-killed antigens in sera of patients with brucellosis.

    PubMed Central

    Araj, G F; Kaufmann, A F

    1989-01-01

    An enzyme-linked immunosorbent assay was used to compare Brucella melitensis major outer membrane proteins (MOMP) and whole-cell heat-killed antigens (HK) in measuring antibrucella immunoglobulin G (IgG), IgM, and IgA in sera of brucellosis patients and controls. Antibodies to MOMP were generally similar to those against HK, and the correlation coefficients between the two antigens and IgG, IgM, and IgA in patients varied between 0.73 and 0.94. Both antigens are comparably suitable in detecting antibrucella immunoglobulin isotypes for the serologic diagnosis of patients with brucellosis, with high (greater than or equal to 95%) sensitivity and specificity. PMID:2768476

  8. Comparing the Thermodynamic Behaviour of Al(1)+ZrO2(s) to Al(1)+Al2O3(s)

    NASA Technical Reports Server (NTRS)

    Copland, Evan

    2004-01-01

    In an effort to better determine the thermodynamic properties of Al(g) and Al2O(g). the vapor in equilibrium with Al(l)+ZrO2(s) was compared to the vapor in equilibrium with Al(l)+Al2O3(s) over temperature range 1197-to-1509K. The comparison was made directly by Knudsen effusion-cell mass spectrometry with an instrument configured for a multiple effusion-cell vapor source (multi-cell KEMS). Second law enthalpies of vaporization of Al(g) and Al2O(g) together with activity measurements show that Al(l)+ZrO2(s) is thermodynamically equivalent to Al(l)+Al2O3(s), indicating Al(l) remained pure and Al2O3(s) was present in the ZrO2-cell. Subsequent observation of the Al(l)/ZrO2 and vapor/ZrO2 interfaces revealed a thin Al2O3-layer had formed, separating the ZrO2-cell from Al(l) and Al(g)+Al2O(g), effectively transforming it into an Al2O3 effusion-cell. This behavior agrees with recent observations made for Beta-NiAl(Pt) alloys measured in ZrO2 effusion-cell.

  9. Ab initio modeling of zincblende AlN layer in Al-AlN-TiN multilayers

    NASA Astrophysics Data System (ADS)

    Yadav, S. K.; Wang, J.; Liu, X.-Y.

    2016-06-01

    An unusual growth mechanism of metastable zincblende AlN thin film by diffusion of nitrogen atoms into Al lattice is established. Using first-principles density functional theory, we studied the possibility of thermodynamic stability of AlN as a zincblende phase due to epitaxial strains and interface effect, which fails to explain the formation of zincblende AlN. We then compared the formation energetics of rocksalt and zincblende AlN in fcc Al through direct diffusion of nitrogen atoms to Al octahedral and tetrahedral interstitials. The formation of a zincblende AlN thin film is determined to be a kinetically driven process, not a thermodynamically driven process.

  10. Differential Antitumor Effects of IgG and IgM Monoclonal Antibodies and Their Synthetic Complementarity-Determining Regions Directed to New Targets of B16F10-Nex2 Melanoma Cells.

    PubMed

    Dobroff, Andrey S; Rodrigues, Elaine G; Juliano, Maria A; Friaça, Dayson M; Nakayasu, Ernesto S; Almeida, Igor C; Mortara, Renato A; Jacysyn, Jacqueline F; Amarante-Mendes, Gustavo P; Magliani, Walter; Conti, Stefania; Polonelli, Luciano; Travassos, Luiz R

    2010-01-01

    Malignant melanoma has increased incidence worldwide and causes most skin cancer-related deaths. A few cell surface antigens that can be targets of antitumor immunotherapy have been characterized in melanoma. This is an expanding field because of the ineffectiveness of conventional cancer therapy for the metastatic form of melanoma. In the present work, antimelanoma monoclonal antibodies (mAbs) were raised against B16F10 cells (subclone Nex4, grown in murine serum), with novel specificities and antitumor effects in vitro and in vivo. MAb A4 (IgG2ak) recognizes a surface antigen on B16F10-Nex2 cells identified as protocadherin beta(13). It is cytotoxic in vitro and in vivo to B16F10-Nex2 cells as well as in vitro to human melanoma cell lines. MAb A4M (IgM) strongly reacted with nuclei of permeabilized murine tumor cells, recognizing histone 1. Although it is not cytotoxic in vitro, similarly with mAb A4, mAb A4M significantly reduced the number of lung nodules in mice challenged intravenously with B16F10-Nex2 cells. The V(H) CDR3 peptide from mAb A4 and V(L) CDR1 and CDR2 from mAb A4M showed significant cytotoxic activities in vitro, leading tumor cells to apoptosis. A cyclic peptide representing A4 CDR H3 competed with mAb A4 for binding to melanoma cells. MAb A4M CDRs L1 and L2 in addition to the antitumor effect also inhibited angiogenesis of human umbilical vein endothelial cells in vitro. As shown in the present work, mAbs A4 and A4M and selected CDR peptides are strong candidates to be developed as drugs for antitumor therapy for invasive melanoma.

  11. Serodiagnosis of Tuberculosis: Comparison of Immunoglobulin A (IgA) Response to Sulfolipid I with IgG and IgM Responses to 2,3-Diacyltrehalose, 2,3,6-Triacyltrehalose, and Cord Factor Antigens

    PubMed Central

    Julián, Esther; Matas, Lurdes; Pérez, Andrés; Alcaide, José; Lanéelle, Marie-Antoinette; Luquin, Marina

    2002-01-01

    Nonpeptidic antigens from the Mycobacterium tuberculosis cell wall are the focus of extensive studies to determine their potential role as protective antigens or serological markers of tuberculous disease. Regarding this latter role and using an enzyme-linked immunosorbent assay, we have made a comparative study of the immunoglobulin G (IgG), IgM, and IgA antibody responses to four trehalose-containing glycolipids purified from M. tuberculosis: diacyltrehaloses, triacyltrehaloses, cord factor, and sulfolipid I (SL-I). Sera from 92 tuberculosis patients (taken before starting antituberculosis treatment) and a wide group of control individuals (84 sera from healthy donors, including purified protein derivative-negative, -positive, healed, and vaccinated individuals, and 52 sera from nontuberculous pneumonia patients), all from Spain, were studied. The results indicated a significantly elevated IgG and IgA antibody response in tuberculosis patients, compared with controls, with all the antigens used. SL-I was the best antigen studied, showing test sensitivities and specificities for IgG of 81 and 77.6%, respectively, and of 66 and 87.5% for IgA. Using this antigen and combining IgA and IgG antibody detection, high test specificity was achieved (93.7%) with a sensitivity of 67.5%. Currently, it is widely accepted that it is not possible to achieve sensitivities above 80% in tuberculosis serodiagnosis when using one antigen alone. Thus, we conclude that SL-I, in combination with other antigenic molecules, could be a useful antigen for tuberculosis serodiagnosis. PMID:12354881

  12. Andalusian astronomy: al-Zij al-Muqtabis of Ibn al-Kammâd.

    NASA Astrophysics Data System (ADS)

    Chabás, J.; Goldstein, B. R.

    1994-12-01

    In the twelfth century the Arabic astronomer Ibn al-Kammâd, living in Islamic Spain, composed three sets of astronomical tables, called "zijes". The zijes were largely based on the work of predecessors, going back to Ptolemy on the one hand, and Hindu astronomers on the other. None of his zijes survived in the original Arabic, but a Latin manuscript contains a translation of an apparently complete zij, the "al-Zij al-Muqtabis". This paper examines the body of astronomical knowledge contained in this zij. Specifically, it is shown that one can gain valuable information on the solar theory of the Islamic astronomers from this translation. Furthermore, the theory of lunar and solar eclipses in the work is analyzed, and tables of mean planetary and solar motions are discussed. In addition, tables for the motion of the vernal equinox, star tables, and several topics of Islamic spherical astronomy are examined.

  13. Microstructure Evolution and Mechanical Properties of Al/Al-Mg/Al composite sheet metals

    NASA Astrophysics Data System (ADS)

    Cho, Jaehyung; Kim, Su-Hyeon; Kim, Hyoung-Wook; Lim, Cha-Yong; Kim, Eun-Young; Choi, Shi-Hoon

    2011-08-01

    Two different types of aluminum alloys of AA1050 and AA5182 were used to manufacture Al/Al-Mg/Al composite sheet metals by roll bonding technology at room temperature. The composite sheet metals were annealed at 400 °C and carried out uniaxial tension tests to investigate mechanical properties. Macroscopic mechanical properties are strongly dependent on the volume (or thickness) fraction of two component layers. Microstructure and texture evolution were also investigated during roll bonding process. The AA1050 sheets located in the outer layer mainly consist of shear texture components and the AA5182 sheet located in the center layer consists of plane strain texture components. With differential speeds of the top and bottom rolls, roll bonding was also carried out. Elongation along the RD and TD was improved at a speed difference of approximately 10%-20%.

  14. Dynamic Modeling of ALS Systems

    NASA Technical Reports Server (NTRS)

    Jones, Harry

    2002-01-01

    The purpose of dynamic modeling and simulation of Advanced Life Support (ALS) systems is to help design them. Static steady state systems analysis provides basic information and is necessary to guide dynamic modeling, but static analysis is not sufficient to design and compare systems. ALS systems must respond to external input variations and internal off-nominal behavior. Buffer sizing, resupply scheduling, failure response, and control system design are aspects of dynamic system design. We develop two dynamic mass flow models and use them in simulations to evaluate systems issues, optimize designs, and make system design trades. One model is of nitrogen leakage in the space station, the other is of a waste processor failure in a regenerative life support system. Most systems analyses are concerned with optimizing the cost/benefit of a system at its nominal steady-state operating point. ALS analysis must go beyond the static steady state to include dynamic system design. All life support systems exhibit behavior that varies over time. ALS systems must respond to equipment operating cycles, repair schedules, and occasional off-nominal behavior or malfunctions. Biological components, such as bioreactors, composters, and food plant growth chambers, usually have operating cycles or other complex time behavior. Buffer sizes, material stocks, and resupply rates determine dynamic system behavior and directly affect system mass and cost. Dynamic simulation is needed to avoid the extremes of costly over-design of buffers and material reserves or system failure due to insufficient buffers and lack of stored material.

  15. Response to Pagano et al.

    PubMed

    Rezende, Flávia; Löwe, Oliver; Helfinger, Valeska; Prior, Kim-Kristin; Walter, Maria; Zukunft, Sven; Fleming, Ingrid; Weissmann, Norbert; Brandes, Ralf P; Schröder, Katrin

    2015-11-20

    In their letter, Pagano et al. appreciate the development of the Nox1, Nox2, and Nox4 triple (3N(-/-)) knockout mouse. They also agree on the view that chemiluminescence assays in general have severe limitations. However, they criticize the fact that the membrane assays in the particular study were restricted to chemiluminescence techniques. Moreover, Pagano et al. got the impression that statements concerning membrane assays of Nox activity in general were made. In addition to a lack of some technical details, Pagano et al. also found the characterization of the 3N(-/-) incomplete and some of the results to be incomprehensible. Although we are grateful for the interest of Pagano et al. in our work, we realized that basically each observation of our study was questioned. This is certainly an excessive rejection of the study in total and fails to appreciate the clear chain of evidences presented. Our work focused on chemiluminescence, and thus, any conclusions are restricted to this technique. Moreover, the 3N(-/-) mice were never developed to study the physiology of Nox enzymes, but rather to validate Nox specificity of NADPH-stimulated chemiluminescence assays. We are convinced that our findings are a valid demonstration that chemiluminescence-based assays in membrane preparations stimulated with NADPH do not measure Nox activity. This conclusion is based on both overexpression studies as well as genetic deficient mouse models. The criticisms of Pagano et al. thus might be justified in some aspects; they, however, cannot disprove the conclusions of our work. Antioxid. Redox Signal. 23, 1247-1249.

  16. Response to Pagano et al.

    PubMed

    Rezende, Flávia; Löwe, Oliver; Helfinger, Valeska; Prior, Kim-Kristin; Walter, Maria; Zukunft, Sven; Fleming, Ingrid; Weissmann, Norbert; Brandes, Ralf P; Schröder, Katrin

    2015-11-20

    In their letter, Pagano et al. appreciate the development of the Nox1, Nox2, and Nox4 triple (3N(-/-)) knockout mouse. They also agree on the view that chemiluminescence assays in general have severe limitations. However, they criticize the fact that the membrane assays in the particular study were restricted to chemiluminescence techniques. Moreover, Pagano et al. got the impression that statements concerning membrane assays of Nox activity in general were made. In addition to a lack of some technical details, Pagano et al. also found the characterization of the 3N(-/-) incomplete and some of the results to be incomprehensible. Although we are grateful for the interest of Pagano et al. in our work, we realized that basically each observation of our study was questioned. This is certainly an excessive rejection of the study in total and fails to appreciate the clear chain of evidences presented. Our work focused on chemiluminescence, and thus, any conclusions are restricted to this technique. Moreover, the 3N(-/-) mice were never developed to study the physiology of Nox enzymes, but rather to validate Nox specificity of NADPH-stimulated chemiluminescence assays. We are convinced that our findings are a valid demonstration that chemiluminescence-based assays in membrane preparations stimulated with NADPH do not measure Nox activity. This conclusion is based on both overexpression studies as well as genetic deficient mouse models. The criticisms of Pagano et al. thus might be justified in some aspects; they, however, cannot disprove the conclusions of our work. Antioxid. Redox Signal. 23, 1247-1249. PMID:26173053

  17. Large rectification magnetoresistance in nonmagnetic Al/Ge/Al heterojunctions.

    PubMed

    Zhang, Kun; Li, Huan-Huan; Grünberg, Peter; Li, Qiang; Ye, Sheng-Tao; Tian, Yu-Feng; Yan, Shi-Shen; Lin, Zhao-Jun; Kang, Shi-Shou; Chen, Yan-Xue; Liu, Guo-Lei; Mei, Liang-Mo

    2015-09-21

    Magnetoresistance and rectification are two fundamental physical properties of heterojunctions and respectively have wide applications in spintronics devices. Being different from the well known various magnetoresistance effects, here we report a brand new large magnetoresistance that can be regarded as rectification magnetoresistance: the application of a pure small sinusoidal alternating-current to the nonmagnetic Al/Ge Schottky heterojunctions can generate a significant direct-current voltage, and this rectification voltage strongly varies with the external magnetic field. We find that the rectification magnetoresistance in Al/Ge Schottky heterojunctions is as large as 250% at room temperature, which is greatly enhanced as compared with the conventional magnetoresistance of 70%. The findings of rectification magnetoresistance open the way to the new nonmagnetic Ge-based spintronics devices of large rectification magnetoresistance at ambient temperature under the alternating-current due to the simultaneous implementation of the rectification and magnetoresistance in the same devices.

  18. Thermodynamic modeling of Pt-Al and Pd-Al

    NASA Astrophysics Data System (ADS)

    Carr, Derek

    2011-03-01

    Pure platinum and pure palladium are too soft for typical jewelry applications. Adding small amounts of other metals can significantly increase their performance. However, international hallmarking standards require the alloys to be 95% pure by weight. How does one achieve significant improvements in performance adding only small amounts (5 wt-%) of other metals? Significant improvements are possible even with small additions if precipitate hardening can be induced. Using a combination of first-principles, cluster expansion, and Monte Carlo modeling, we have identified new Pt-rich/Pd-rich phases in Pt-Al and Pd-Al that should be useful in precipitate hardening. Thermodynamical modeling indicates that the phases are experimentally feasible (not kinetically inhibited).

  19. Large rectification magnetoresistance in nonmagnetic Al/Ge/Al heterojunctions

    NASA Astrophysics Data System (ADS)

    Zhang, Kun; Li, Huan-Huan; Grünberg, Peter; Li, Qiang; Ye, Sheng-Tao; Tian, Yu-Feng; Yan, Shi-Shen; Lin, Zhao-Jun; Kang, Shi-Shou; Chen, Yan-Xue; Liu, Guo-Lei; Mei, Liang-Mo

    2015-09-01

    Magnetoresistance and rectification are two fundamental physical properties of heterojunctions and respectively have wide applications in spintronics devices. Being different from the well known various magnetoresistance effects, here we report a brand new large magnetoresistance that can be regarded as rectification magnetoresistance: the application of a pure small sinusoidal alternating-current to the nonmagnetic Al/Ge Schottky heterojunctions can generate a significant direct-current voltage, and this rectification voltage strongly varies with the external magnetic field. We find that the rectification magnetoresistance in Al/Ge Schottky heterojunctions is as large as 250% at room temperature, which is greatly enhanced as compared with the conventional magnetoresistance of 70%. The findings of rectification magnetoresistance open the way to the new nonmagnetic Ge-based spintronics devices of large rectification magnetoresistance at ambient temperature under the alternating-current due to the simultaneous implementation of the rectification and magnetoresistance in the same devices.

  20. AlSb/InAs/AlSb quantum wells

    NASA Technical Reports Server (NTRS)

    Kroemer, Herbert

    1990-01-01

    Researchers studied the InAs/AlSb system recently, obtaining 12nm wide quantum wells with room temperature mobilities up to 28,000 cm(exp 2)/V center dot S and low-temperature mobilities up to 325,000 cm(exp 2)/V center dot S, both at high electron sheet concentrations in the 10(exp 12)/cm(exp 2) range (corresponding to volume concentrations in the 10(exp 18)/cm(exp 2) range). These wells were not intentionally doped; the combination of high carrier concentrations and high mobilities suggest that the electrons are due to not-intentional modulation doping by an unknown donor in the AlSb barriers, presumably a stoichiometric defect, like an antisite donor. Inasmuch as not intentionally doped bulk AlSb is semi-insulating, the donor must be a deep one, being ionized only by draining into the even deeper InAs quantum well. The excellent transport properties are confirmed by other observations, like excellent quantum Hall effect data, and the successful use of the quantum wells as superconductive weak links between Nb electrodes, with unprecendentedly high critical current densities. The system is promising for future field effect transistors (FETs), but many processing problems must first be solved. Although the researchers have achieved FETs, the results so far have not been competitive with GaAs FETs.

  1. Identification of Candida albicans ALS2 and ALS4 and Localization of Als Proteins to the Fungal Cell Surface

    PubMed Central

    Hoyer, L. L.; Payne, T. L.; Hecht, J. E.

    1998-01-01

    Additional genes in the growing ALS family of Candida albicans were isolated by PCR screening of a genomic fosmid library with primers designed from the consensus tandem-repeat sequence of ALS1. This procedure yielded fosmids encoding ALS2 and ALS4. ALS2 and ALS4 conformed to the three-domain structure of ALS genes, which consists of a central domain of tandemly repeated copies of a 108-bp motif, an upstream domain of highly conserved sequences, and a domain of divergent sequences 3′ of the tandem repeats. Alignment of five predicted Als protein sequences indicated conservation of N- and C-terminal hydrophobic regions which have the hallmarks of secretory signal sequences and glycosylphosphatidylinositol addition sites, respectively. Heterologous expression of an N-terminal fragment of Als1p in Saccharomyces cerevisiae demonstrated function of the putative signal sequence with cleavage following Ala17. This signal sequence cleavage site was conserved in the four other Als proteins analyzed, suggesting identical processing of each protein. Primary-structure features of the five Als proteins suggested a cell-surface localization, which was confirmed by indirect immunofluorescence with an anti-Als antiserum. Staining was observed on mother yeasts and germ tubes, although the intensity of staining on the mother yeast decreased with elongation of the germ tube. Similar to other ALS genes, ALS2 and ALS4 were differentially regulated. ALS4 expression was correlated with the growth phase of the culture; ALS2 expression was not observed under many different in vitro growth conditions. The data presented here demonstrate that ALS genes encode cell-surface proteins and support the conclusion that the size and number of Als proteins on the C. albicans cell surface vary with strain and growth conditions. PMID:9765564

  2. Novel acid-labile subunit ( IGFALS ) mutation p.T145K (c.434C>A) in a patient with ALS deficiency, normal stature and immunological dysfunction.

    PubMed

    Schreiner, Felix; Schoenberger, Stefan; Koester, Bernhard; Domené, Horacio M; Woelfle, Joachim

    2013-01-01

    We report a novel missense mutation p.T145K in the insulin-like growth factor (IGF) acid-labile subunit (IGFALS) gene identified in a Turkish patient with normal growth, transient pancytopenic episodes and signs of immunological dysfunction. Because of recurrent cutaneous mycoses and absence of pubertal development until the age of 14.75 years we determined several endocrine parameters in order to rule out autoimmune-polyendocrine syndromes. Despite a normal height between the 25th and 50th percentile we found severely decreased IGF-1 and undetectably low IGFBP-3 levels. Laboratory signs of immunological dysfunction included reduced total lymphocyte count with diminished B and T helper cell fractions, decreased serum concentrations of IgM and IgG subclass 4, and elevated antinuclear antibody and anti-dsDNA titers as well as persistently high interleukin-2-receptor levels. Further endocrine work-up revealed elevated fasting insulin and undetectably low ALS serum levels, leading to the diagnosis of ALS deficiency. Sequencing of the coding region of the IGFALS gene showed a novel homozygous missense mutation (c.434C>A; p.T145K). Since immunological abnormalities have not been reported in more than 20 ALS-deficient patients so far and our patient was born to consanguineous parents, a second autosomal recessive defect is likely to underlie the immunological phenotype, although a causative role of IGFALS p.T145K cannot be entirely ruled out. PMID:24296365

  3. Establishment of an Algorithm Using prM/E- and NS1-Specific IgM Antibody-Capture Enzyme-Linked Immunosorbent Assays in Diagnosis of Japanese Encephalitis Virus and West Nile Virus Infections in Humans

    PubMed Central

    Galula, Jedhan U.; Chang, Gwong-Jen J.

    2015-01-01

    The front-line assay for the presumptive serodiagnosis of acute Japanese encephalitis virus (JEV) and West Nile virus (WNV) infections is the premembrane/envelope (prM/E)-specific IgM antibody-capture enzyme-linked immunosorbent assay (MAC-ELISA). Due to antibody cross-reactivity, MAC-ELISA-positive samples may be confirmed with a time-consuming plaque reduction neutralization test (PRNT). In the present study, we applied a previously developed anti-nonstructural protein 1 (NS1)-specific MAC-ELISA (NS1-MAC-ELISA) on archived acute-phase serum specimens from patients with confirmed JEV and WNV infections and compared the results with prM/E containing virus-like particle-specific MAC-ELISA (VLP-MAC-ELISA). Paired-receiver operating characteristic (ROC) curve analyses revealed no statistical differences in the overall assay performances of the VLP- and NS1-MAC-ELISAs. The two methods had high sensitivities of 100% but slightly lower specificities that ranged between 80% and 100%. When the NS1-MAC-ELISA was used to confirm positive results in the VLP-MAC-ELISA, the specificity of serodiagnosis, especially for JEV infection, was increased to 90% when applied in areas where JEV cocirculates with WNV, or to 100% when applied in areas that were endemic for JEV. The results also showed that using multiple antigens could resolve the cross-reactivity in the assays. Significantly higher positive-to-negative (P/N) values were consistently obtained with the homologous antigens than those with the heterologous antigens. JEV or WNV was reliably identified as the currently infecting flavivirus by a higher ratio of JEV-to-WNV P/N values or vice versa. In summary of the above-described results, the diagnostic algorithm combining the use of multiantigen VLP- and NS1-MAC-ELISAs was developed and can be practically applied to obtain a more specific and reliable result for the serodiagnosis of JEV and WNV infections without the need for PRNT. The developed algorithm should provide great

  4. Al-Anon and recovery.

    PubMed

    Cermak, T L

    1989-01-01

    The history of Al-Anon and its current demographics are reviewed. In order to understand 12-step recovery and psychotherapy for family members of alcoholics, the concept of codependence is defined with a set of diagnostic criteria consistent with the DSM-III-R definition of personality traits and disorders. At the core of codependence are denial and an unrealistic relationship to willpower. The therapeutic implications of considering codependence as a personality disorder are explored, as are the characteristics that make codependence unique among personality disorders: the central role of denial and the existence of a self-help organization to facilitate recovery. The dynamics of working the 12 steps on codependent characteristics are outlined. A synergistic relationship between psychotherapy and the 12 steps is described. Special attention is given the emergence of Al-Anon adult children of alcoholic meetings, and the future of codependence is discussed.

  5. Emerging mechanisms of molecular pathology in ALS.

    PubMed

    Peters, Owen M; Ghasemi, Mehdi; Brown, Robert H

    2015-05-01

    Amyotrophic lateral sclerosis (ALS) is a devastating degenerative disease characterized by progressive loss of motor neurons in the motor cortex, brainstem, and spinal cord. Although defined as a motor disorder, ALS can arise concurrently with frontotemporal lobal dementia (FTLD). ALS begins focally but disseminates to cause paralysis and death. About 10% of ALS cases are caused by gene mutations, and more than 40 ALS-associated genes have been identified. While important questions about the biology of this disease remain unanswered, investigations of ALS genes have delineated pathogenic roles for (a) perturbations in protein stability and degradation, (b) altered homeostasis of critical RNA- and DNA-binding proteins, (c) impaired cytoskeleton function, and (d) non-neuronal cells as modifiers of the ALS phenotype. The rapidity of progress in ALS genetics and the subsequent acquisition of insights into the molecular biology of these genes provide grounds for optimism that meaningful therapies for ALS are attainable. PMID:25932674

  6. Synthesis of AlN/Al Polycrystals along with Al Nanoparticles Using Thermal Plasma Route

    SciTech Connect

    Kanhe, Nilesh S.; Nawale, A. B.; Kulkarni, N. V.; Bhoraskar, S. V.; Mathe, V. L.; Das, A. K.

    2011-07-15

    This paper for the first time reports the (200) oriented growth of hexagonal Aluminum nitride crystals during synthesis of aluminum nanoparticles in dc transferred arc thermal plasma reactor by gas phase condensation in nitrogen plasma. The structural and morphological study of as synthesized AlN crystal and aluminium nanoparticles was done by using the x-ray diffraction method, scanning electron microscopy and transmission electron microscopy.

  7. Ab initio modeling of zincblende AlN layer in Al-AlN-TiN multilayers

    DOE PAGES

    Yadav, S. K.; Wang, J.; Liu, X. -Y.

    2016-06-13

    An unusual growth mechanism of metastable zincblende AlN thin film by diffusion of nitrogen atoms into Al lattice is established. Using first-principles density functional theory, we studied the possibility of thermodynamic stability of AlN as a zincblende phase due to epitaxial strains and interface effect, which fails to explain the formation of zincblende AlN. We then compared the formation energetics of rocksalt and zincblende AlN in fcc Al through direct diffusion of nitrogen atoms to Al octahedral and tetrahedral interstitials. Furthermore, the formation of a zincblende AlN thin film is determined to be a kinetically driven process, not a thermodynamicallymore » driven process.« less

  8. Synthesis of Al-Al2O3 and Al-Aln Nanoparticle Composites Via Electric Explosion of Wires

    NASA Astrophysics Data System (ADS)

    Lerner, M. I.; Lozhkomoev, A. S.; Pervikov, A. V.; Bakina, O. V.

    2016-07-01

    Composite Al-Al2O3 and Al-AlN nanoparticles were synthesized via electric explosion of aluminum wires in an argon-oxygen gas mixture and in nitrogen. The parameters of electric explosion and gas medium affect the size and relative content of nitride and aluminum oxide in the nanoparticles. Processes of forming chemical compounds during aluminum oxidation at the contact surface between explosive products and gas and of nitrogen diffusions into the nanoparticles of the condensed phase are considered.

  9. NiAl-base composite containing high volume fraction of AlN for advanced engines

    NASA Technical Reports Server (NTRS)

    Hebsur, Mohan (Inventor); Whittenbeger, John D. (Inventor); Lowell, Carl F. (Inventor)

    1994-01-01

    A particulate reinforced NiAl-AlN composite alloy has a NiAl matrix and greater than about 13 volume percent fine particles of AlN within the matrix. The particles preferably have a diameter from about 15 nanometers to about 50 nanometers. The particulate reinforced NiAl-AlN composite alloy may be prepared by cryomilling prealloyed NiAl in liquid nitrogen using grinding media having a diameter of from about 2 to 6 mm at an impeller speed of from about 450 RPM to about 800 RPM. The cryomilling may be done for a duration of from about 4 hours to about 20 hours to obtain a cryomilled powder. The cryomilled powder may be consolidated to form the particulate reinforced NiAl-AlN composite alloy. The particulate reinforced alloy can further include a toughening alloy. The toughening alloy may include NiCrAlY, FeCrAlY, and FeAl.

  10. [Effects of the use of FRIGEN (AG-Ffm-HOECST) on the determination of blood levels of some proteins (IgA, IgM, IgG, C3c, C4) in cord blood of healthy newborn infants at term by laser nephelometry].

    PubMed

    Costanzo, A; Meninno, V; Addeo, L; Canzano, G; Lombardi, S

    1984-09-30

    The purpose of this paper is to establishes the FRIGEN effects on the determination of IgA, IgM, IgG, C3c, C4 cord-blood levels, by means of laser-nephelometry. The results show substantial interferences only in the IgA levels, whereas the other on almost all proteins are not affected, the found interference can be due to the amount of IgA bound to B-lipoprotein, that are precipitated by FRIGEN.

  11. Reply to Pachai et al.

    PubMed

    Harrison, William J; Bex, Peter J

    2016-05-01

    Peripheral vision is fundamentally limited by the spacing between objects. When asked to report a target's identity, observers make erroneous reports that sometimes match the identity of a nearby distractor and sometimes match a combination of target and distractor features. The classification of these errors has previously been used to support competing 'substitution' [1] or 'averaging' [2] models of the phenomenon known as 'visual crowding'. We recently proposed a single model in which both classes of error occur because observers make their reports by sampling from a biologically-plausible population of weighted responses within a region of space around the target [3]. It is critical to note that there is no probabilistic substitution or averaging process in our model; instead, we argue that neither substitution nor averaging occur, but that these are misclassifications of the distribution of reports that emerge when a population response distribution is sampled. This is a fundamentally different way of thinking about crowding, and on this basis we claim to have provided a mechanism unifying categorically distinct perceptual errors. Our goal was not to model all crowding phenomena, such as the release from crowding when target and flanks differ in color or depth [4]. Pachai et al.[5] have suggested that our model is not unifying because it inaccurately predicts perceptual performance for a particular stimulus. Although we agree that our model does not predict their data, this specific demonstration overlooks the critical aspect of the model: perceptual reports are drawn from a weighted population code. We show that Pachai et al.'s [5] own data actually provide evidence for the population code we have described [3], and we suggest a biologically-plausible analysis of their stimuli that provides a computational basis for their 'grouping' account of crowding. PMID:27166690

  12. Reply to Pachai et al.

    PubMed

    Harrison, William J; Bex, Peter J

    2016-05-01

    Peripheral vision is fundamentally limited by the spacing between objects. When asked to report a target's identity, observers make erroneous reports that sometimes match the identity of a nearby distractor and sometimes match a combination of target and distractor features. The classification of these errors has previously been used to support competing 'substitution' [1] or 'averaging' [2] models of the phenomenon known as 'visual crowding'. We recently proposed a single model in which both classes of error occur because observers make their reports by sampling from a biologically-plausible population of weighted responses within a region of space around the target [3]. It is critical to note that there is no probabilistic substitution or averaging process in our model; instead, we argue that neither substitution nor averaging occur, but that these are misclassifications of the distribution of reports that emerge when a population response distribution is sampled. This is a fundamentally different way of thinking about crowding, and on this basis we claim to have provided a mechanism unifying categorically distinct perceptual errors. Our goal was not to model all crowding phenomena, such as the release from crowding when target and flanks differ in color or depth [4]. Pachai et al.[5] have suggested that our model is not unifying because it inaccurately predicts perceptual performance for a particular stimulus. Although we agree that our model does not predict their data, this specific demonstration overlooks the critical aspect of the model: perceptual reports are drawn from a weighted population code. We show that Pachai et al.'s [5] own data actually provide evidence for the population code we have described [3], and we suggest a biologically-plausible analysis of their stimuli that provides a computational basis for their 'grouping' account of crowding.

  13. Everyday Life with ALS: A Practical Guide

    MedlinePlus

    ... course of ALS. ALS also will require some adaptations to your environment, both for safety and to ... the way, Chapter 6 will include information on adaptations for computer access. Though the chapter is called “ ...

  14. Effects of the Al content on pore structures of porous TieAl alloys

    SciTech Connect

    Jaing, Y; He, Y H; Xu, N P; Zou, J; Huang, B; Lui, C T

    2008-01-01

    Porous TieAl alloys with different nominal compositions were fabricated through a reactive synthesis of Ti and Al elemental powders. It has been found that the pore parameters vary with the Al contents, indicating that the nature of the pores can be manipulated through changing the Al contents. In addition, detailed structural characterizations showed that the fabricated porous TieAl alloys can have three crystalline phases (i.e., a2-Ti3Al, g-TiAl, and TiAl3) when using different compositions. The fundamental reasons behind these phenomena have been explored.

  15. Nonstoichiometry of Al-Zr intermetallic phases

    SciTech Connect

    Radmilovic, V.; Thomas, G.

    1994-06-01

    Nonstoichiometry of metastable cubic {beta}{prime} and equilibrium tetragonal {beta} Al-Zr intermetallic phases of the nominal composition Al{sub 3}Zr in Al-rich alloys has been extensively studied. It is proposed that the ``dark contrast`` of {beta}{prime} core in {beta}{prime}/{sigma}{prime} complex precipitates, in Al-Li-Zr based alloys, is caused by incorporation of Al and Li atoms into the {beta}{prime} phase on Zr sublattice sites, forming nonstoichiometric Al-Zr intermetallic phases, rather than by Li partitioning only. {beta}{prime} particles contain very small amounts of Zr, approximately 5 at.%, much less than the stoichiometric 25 at.% in the Al{sub 3}Zr metastable phase. These particles are, according to simulation of high resolution images, of the Al{sub 3}(Al{sub 0.4}Li{sub 0.4}Zr{sub 0.2}) type. Nonstoichiometric particles of average composition Al{sub 4}Zr and Al{sub 6}Zr are observed also in the binary Al-Zr alloy, even after annealing for several hours at 600{degree}C.

  16. Epitope expression and partial structural characterization of F62 lipooligosaccharide (LOS) of Neisseria gonorrhoeae: IgM monoclonal antibodies (3F11 and 1-1-M) recognize non-reducing termini of the LOS components.

    PubMed

    Yamasaki, R; Nasholds, W; Schneider, H; Apicella, M A

    1991-11-01

    F62 LOS of Neisseria gonorrhoeae consists of two components. The higher molecular weight (MW) component is recognized by monoclonal antibody (MAb) 1-1-M and the smaller MW component by MAb 3F11. Epitope expression of the two LOS components and their partial structures were investigated by treating the F62 LOS with several glycosidases and then monitoring their antigenicity with the two mouse IgM MAbs. The 1-1-M-defined LOS component was cleaved with both beta-N-acetylhexosaminidase and endo-beta-galactosidase, and each cleavage resulted in the loss of expression of the 1-1-M-defined epitope. The N-acetylhexosamine (HexNAc) released by the hexosaminidase was found to be GalNAc, and the smaller oligosaccharide released by the endo enzyme was identified to be a dimer GalNAc beta----Gal. In contrast, the MAb 3F11-defined LOS component was not digested by the endo galactosidase, but it was cleaved with alpha and beta-galactosidase, and expression of the MAb 3F11-defined LOS epitope expression of the MAb 3F11-defined LOS was abolished by the treatment with each of two exo enzymes. MAb 3F11 bound to the 1-1-M-defined LOS component resulting from the removal of the beta-GalNAc residue, and the resulting LOS was further cleaved with beta-galactosidase, but not with alpha-galactosidase. From these results, we conclude the following: (1) MAbs 1-1-M and 3F11 both recognize the non-reducing termini of the LOS components; (2) the 1-1-M-defined LOS component has the GalNAc beta----Gal beta 1----4-Glc (or GlcNAc) structure, and the GalNAc beta----Gal residue is involved in the MAb 1-1-M-defined epitope; (3) the MAb 3F11-defined LOS component may not have a Gal beta 1----4GlcNAc beta 1----4Gal beta 1----4Glc structure within the molecule. However, it has beta-Gal residue at its non-reducing terminus, and this residue is involved in the MAb 3F11-defined epitope; (4) the two LOS components share a similar antigenic structure, and the 3F11-defined epitope structure is present in the

  17. Cenomanian-Turonian biostratigraphy of the Jardas Al Abid area, Al Jabal Al Akhdar, northeast Libya

    NASA Astrophysics Data System (ADS)

    El Qot, Gamal M.; Abdulsamad, Esam O.

    2016-09-01

    The Upper Cenomanian-Turonian succession exposed at Jardas al'Abid area consists mainly of carbonates with siliciclastic intercalations. This succession is subdivided lithostratigraphically into: Qasr al'Abid (Late Cenomanian) and Al Baniyah (Late Cenomanian-Coniacian) formations. This sequence is relatively rich in macrofossil assemblages especially bivalves, gastropods, and echinoids with rare ammonites. Based on the first occurrence (FO) and last occurrence (LO) of some index species of these macrofossil groups, an integrated biostratigraphic framework has been constructed. The studied Cenomanian-Turonian sequence is subdivided biostratigraphically into three ammonite biozones; Pseudaspidoceras pseudonodosoides Total Range Zone, Choffaticeras segne Total Range Zone, and Coilopoceras requienianum Total Range Zone. Based on the rest of macrofossil assemblages other than the ammonites, eight biozones were recognized; Mecaster batnensis Total Range Zone, Ceratostreon flabellatum-Neithea dutrugei Acme Zone, Costagyra olisiponensis Acme Zone, Pycnodonte (Phygraea) vesicularis vesiculosa Acme Zone, Mytiloides labiatus Total Range Zone = Mecaster turonensis Acme Zone, Rachiosoma rectilineatum-Curvostrea rouvillei-Tylostoma (T.) globosum Assemblage Zone, Radiolites sp.-Apricardia? matheroni Total Range Zone, and Nerinea requieniana Total Range Zone. Most of the proposed biozones are recorded for the first time from Libya. The integration among these biozones as well as local and inter-regional correlation of the biozones have been discussed. The stage boundaries of the studied stratigraphic intervals are discussed, where the Cenomanian/Turonian boundary is delineated at the last occurrence (LO) of Pseudaspidoceras pseudonodosoides (Choffat), while the Turonian/Coniacian boundary is delineated arbitrary being agree with the LO of the Turonian fauna.

  18. Cyclic oxidation resistance of a reaction milled NiAl-AlN composite

    NASA Technical Reports Server (NTRS)

    Lowell, Carl E.; Barrett, Charles A.; Whittenberger, J. D.

    1990-01-01

    Based upon recent mechanical property tests a NiAl-AlN composite produced by cryomilling has very attractive high temperature strength. This paper focuses on the oxidation resistance of the NiAl-AlN composite at 1473 and 1573 K as compared to that of Ni-47Al-0.15Zr, one of the most oxidation resistant intermetallics. The results of cyclic oxidation tests show that the NiAl-AlN composite has excellent properties although not quite as good as those of Ni-47Al-0.15Zr. The onset of failure of the NiAl-AlN was unique in that it was not accompanied by a change in scale composition from alumina to less protective oxides. Failure in the composite appears to be related to the entrapment of AlN particles within the alumina scale.

  19. Wear behavior of Al-Al{sub 3}Ti composite manufactured by a centrifugal method

    SciTech Connect

    Watanabe, Yoshimi; Yamanaka, Noboru; Fukui, Yasuyoshi

    1999-12-01

    The purpose of the present study was to develop a wear-resistant, light Al-Al{sub 3}Ti composite material. An Al-Al{sub 3}Ti composite specimen was machined from a thick-walled tube of Al-Al{sub 3}Ti functionally graded material (FGM) manufactured by the centrifugal method from a commercial ingot of Al-5 mass% Ti master alloy. The alloy was heated to a temperature where solid Al{sub 3}Ti particles resided in a liquid Al matrix, and then the centrifugal method was carried out. Al{sub 3}Ti particles in a commercial alloy ingot exist as platelets, and this shape was maintained through the casting. Three kinds of wear specimens were prepared, taking into account the morphology of the Al{sub 3}Ti particles in the thick-walled FGM tube; the Al{sub 3}Ti particles were arranged with their platelet planes nearly normal to the radial direction as a result of the applied centrifugal force. The wear resistance of the Al-Al{sub 3}Ti composite was significantly higher than that of pure Al. Wear-resistance anisotropy and dissolution of the Al{sub 3}Ti into the Al matrix at the near-surface region, around 100 {micro}m in depth, were also observed. The mechanism of the supersaturated-layer formation and the origin of the anisotropic wear resistance are discussed.

  20. Systems Engineering Techniques for ALS Decision Making

    NASA Technical Reports Server (NTRS)

    Rodriquez, Luis F.; Drysdale, Alan E.; Jones, Harry; Levri, Julie A.

    2004-01-01

    The Advanced Life Support (ALS) Metric is the predominant tool for predicting the cost of ALS systems. Metric goals for the ALS Program are daunting, requiring a threefold increase in the ALS Metric by 2010. Confounding the problem, the rate new ALS technologies reach the maturity required for consideration in the ALS Metric and the rate at which new configurations are developed is slow, limiting the search space and potentially giving the perspective of a ALS technology, the ALS Metric may remain elusive. This paper is a sequel to a paper published in the proceedings of the 2003 ICES conference entitled, "Managing to the metric: an approach to optimizing life support costs." The conclusions of that paper state that the largest contributors to the ALS Metric should be targeted by ALS researchers and management for maximum metric reductions. Certainly, these areas potentially offer large potential benefits to future ALS missions; however, the ALS Metric is not the only decision-making tool available to the community. To facilitate decision-making within the ALS community a combination of metrics should be utilized, such as the Equivalent System Mass (ESM)-based ALS metric, but also those available through techniques such as life cycle costing and faithful consideration of the sensitivity of the assumed models and data. Often a lack of data is cited as the reason why these techniques are not considered for utilization. An existing database development effort within the ALS community, known as OPIS, may provide the opportunity to collect the necessary information to enable the proposed systems analyses. A review of these additional analysis techniques is provided, focusing on the data necessary to enable these. The discussion is concluded by proposing how the data may be utilized by analysts in the future.

  1. Instandhaltungsmanagement als Gestaltungsfeld Ganzheitlicher Produktionssysteme

    NASA Astrophysics Data System (ADS)

    Dombrowski, Uwe; Schulze, Sven; Otano, Isabel Crespo

    Sich kontinuierlich verändernde Rahmenbedingungen, wie beispielsweise eine steigende Variantenvielfalt, verkürzte Produktlebenszyklen sowie Kundenforderungen nach höherer Qualität, kürzeren Lieferzeiten und geringeren Kosten, fordern von produzierenden Unternehmen eine stetige Anpassung der Prozesse, der Organisation und der Strukturen. Seit den 90er Jahren versuchen immer mehr deutsche Unternehmen diesen veränderten Anforderungen mit der Einführung eines Ganzheitlichen Produktionssystems (GPS) zu begegnen. Ganzheitliche Produktionssysteme sind dabei in ihren Grundlagen an das Toyota Produktionssystem angelehnt, vereinigen aber auch weitere Methoden zu einem unternehmensspezifischen Regelwerk. Im Rahmen des langfristigen Trends zu unternehmensindividuellen Produktionssystemen wird sowohl in der Industrie als auch in der Forschung intensiv über das Toyota Produktionssystem, Lean Production, Lean Management und Ganzheitliche Produktionssysteme diskutiert, werden Konzepte zu Implementierung und Betrieb erstellt und die Wirtschaftlichkeit untersucht.

  2. Structure and stability of the AlX and AlX{sup {minus}} species

    SciTech Connect

    Gutsev, G.L.; Jena, P.; Bartlett, R.J.

    1999-02-01

    The electronic and geometrical structures of the ground and low-lying excited states of the diatomic AlX and AlX{sup {minus}} series (X=H, Li, Be, B, C, N, O, and F) are calculated by the coupled-cluster method with all singles and doubles and noniterative inclusion of triples using a large atomic natural orbital basis. All the ground-state AlX molecules except for AlF can attach an additional electron and form ground-state AlX{sup {minus}} anions. The ground-state AlBe{sup {minus}}, AlB{sup {minus}}, AlC{sup {minus}}, AlN{sup {minus}}, and AlO{sup {minus}} anions possess excited states that are stable toward autodetachment of an extra electron; AlBe{sup {minus}} also has a second excited state. Low-lying excited states of all AlX but AlN can attach an extra electron and form anionic states that are stable with respect to their neutral (excited) parent states. The ground-state AlLi{sup {minus}}, AlBe{sup {minus}}, AlB{sup {minus}}, AlN{sup {minus}}, and AlO{sup {minus}} anions are found to be thermodynamically more stable than their neutral parents. The most stable is AlO{sup {minus}}, whose dissociation energy to Al+O{sup {minus}} is 6.4 eV. Correspondingly, AlO possesses the largest electron affinity (2.65 eV) in the series. {copyright} {ital 1999 American Institute of Physics.}

  3. Accumulation of Al in Root Mucilage of an Al-Resistant and an Al-Sensitive Cultivar of Wheat.

    PubMed Central

    Archambault, D. J.; Zhang, G.; Taylor, G. J.

    1996-01-01

    To estimate rates of Al accumulation within the symplasm, all apoplastic pools of Al need to be eliminated or accounted for. We have developed a revised kinetic protocol that allows us to estimate the contribution of mucilage-bound Al to total, nonexchangeable Al, and to eliminate the mucilage as an apoplastic pool of Al. By comparing the Al content of excised root tips (2 cm) of wheat (Triticum aestivum L.) with and without the removal of the mucilage (using a 10-min wash in 1 M NH4Cl), we found that Al bound to the mucilage accounted for approximately 25 to 35% of Al remaining after desorption in citric acid. The kinetics of Al uptake into mucilage were biphasic, with a rapid phase occurring in the first 30 min of uptake, followed by a linear phase occurring in the remainder of the experimental period (180 min). By adopting a step for removal of mucilage into our existing kinetic protocol, we have been able to isolate a linear phase of uptake with only a slight deviation from linearity in the first 5 min. Although we cannot unambiguously identify this phase of uptake as uptake into the symplasm, we believe this new protocol provides us with the most accurate quantitative estimate of symplastic Al yet available. PMID:12226458

  4. Magnetism of Al-substituted magnetite reduced from Al-hematite

    NASA Astrophysics Data System (ADS)

    Jiang, Zhaoxia; Liu, Qingsong; Zhao, Xiang; Roberts, Andrew P.; Heslop, David; Barrón, Vidal; Torrent, José

    2016-06-01

    Aluminum-substituted magnetite (Al-magnetite) reduced from Al-substituted hematite or goethite (Al-hematite or Al-goethite) is an environmentally important constituent of magnetically enhanced soils. In order to characterize the magnetic properties of Al-magnetite, two series of Al-magnetite samples were synthesized through reduction of Al-hematite by a mixed gas (80% CO2 and 20% CO) at 395°C for 72 h in a quartz tube furnace. Al-magnetite samples inherited the morphology of their parent Al-hematite samples, but only those transformed from Al-hematite synthesized at low temperature possessed surficial micropores, which originated from the release of structural water during heating. Surface micropores could thus serve as a practical fingerprint of fire or other high-temperature mineralogical alteration processes in natural environments, e.g., shear friction in seismic zones. In addition, Al substitution greatly affects the magnetic properties of Al-magnetite. For example, coercivity (Bc) increases with increasing Al content and then decreases slightly, while the saturation magnetization (Ms), Curie temperature (Tc), and Verwey transition temperature (Tv) all decrease with increasing Al content due to crystal defect formation and dilution of magnetic ions caused by Al incorporation. Moreover, different trends in the correlation between Tc and Bc can be used to discriminate titanomagnetite from Al-magnetite, which is likely to be important in environmental and paleomagnetic studies, particularly in soil.

  5. Reactive Plasma Spraying of Fine Al2O3/AlN Feedstock Powder

    NASA Astrophysics Data System (ADS)

    Shahien, Mohammed; Yamada, Motohiro; Yasui, Toshiaki; Fukumoto, Masahiro

    2013-12-01

    Reactive plasma spraying (RPS) is a promising technology for in situ formation of aluminum nitride (AlN) coatings. Recently, AlN-based coatings were fabricated by RPS of alumina (Al2O3) powder in N2/H2 thermal plasma. This study investigated the feasibility of RPS of a fine Al2O3/AlN mixture and the influence of the plasma gases (N2, H2) on the nitriding conversion, and coating microstructure and properties. Thick AlN/Al2O3 coatings with high nitride content were successfully fabricated. The coatings consist of h-AlN, c-AlN, Al5O6N, γ-Al2O3, and a small amount of α-Al2O3. Use of fine particles enhanced the nitriding conversion and the melting tendency by increasing the surface area. Furthermore, the AlN additive improved the AlN content in the coatings. Increasing the N2 gas flow rate improved the nitride content and complete crystal growth to the h-AlN phase, and enhanced the coating thickness. On the other hand, though the H2 gas is required for plasma nitriding of the Al2O3 particles, increasing its flow rate decreased the nitride content and the coating thickness. Remarkable influence of the plasma gases on the coating composition, microstructure, and properties was observed during RPS of the fine particles.

  6. 77 FR 73732 - In the Matter of Amendment of the Designation of al-Qa'ida in Iraq, aka Jam'at al Tawhid wa'al...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-12-11

    ... al-Rafidayn, aka The Organization of al-Jihad's Base of Operations in Iraq, aka al-Qaida of Jihad in Iraq, aka al-Qaida in Iraq, aka al-Qaida in Mesopotamia, aka al-Qaida in the Land of the Two Rivers, aka al-Qaida of the Jihad in the Land of the Two Rivers, aka al-Qaida of Jihad Organization in......

  7. A comparison of mechanical properties between Al and Al3Mg

    NASA Astrophysics Data System (ADS)

    Yang, Rong; Tang, Bin; Gao, Tao

    2016-11-01

    On the basis of first principles calculations, we have calculated the elastic properties, stress-strain relations, ideal tensile strengths, ideal shear strengths, and the ideal compressed strengths of Al and Al3Mg. The stress-strain relations of Al3Mg are strikingly similar to those of Al, indicating that the crystal structure appears to be more important than the identity of the individual atoms during uniaxial deformation. Al3Mg is found to have larger moduli and higher strengths than Al but less ductile than Al. So Al3Mg is expected to be a harder material, consistent with its exploitation in Al precipitate-hardening mechanisms. The calculated elastic properties, tensile strengths and shear strengths of Al are consistent with experimental values or previous theoretical results. We also use another method (molecular dynamics (MD) simulations) to recalculate elastic constants, ideal tensile and compressed strength of Al3Mg for checking and comparing. We find that the results obtained by the two methods agree well with each other. The failure modes under uniaxial <100> tension are also explored for Al and Al3Mg. Our calculations confirm that Al fail by shear and predict that Al3Mg also fail by shear.

  8. Influences of Al particles on the microstructure and property of electrodeposited Ni-Al composite coatings

    NASA Astrophysics Data System (ADS)

    Cai, Fei; Jiang, Chuanhai

    2014-02-01

    Ni-Al composite coatings with different contents of Al microparticles were prepared from a conventional Watt bath. The influences of Al particle loadings in the bath on the surface morphology, composition, texture, grain size, microstrain, residual stress and anti-corrosion of the Ni-Al composite coating were investigated. The friction coefficients of the coatings at 200 °C were also evaluated by a pin-on-disctribometer. The results showed that the surface morphology of the coatings changed from pyramid + colonied structure to colonied structure with increasing Al particle loadings. The (2 0 0) preferred orientation for pure Ni coating evolved to random orientation with increasing Al particle loadings. The grain size obtained the minimum value of 72.28 nm at Al particle loading of 100 g/L and the microstrain of the coating increased with increasing the Al particle loadings. The incorporation of Al particles decreased the residual stress of the electro-deposited coating and all the coatings deposited at different Al particle loadings possessed low residual stress. As the Al particle loading increased, the anti-corrosion of the Ni-Al coatings increased owing to the combined effect of increasing Al content in the coatings and the texture evolution from (2 0 0) plane to (1 1 1) plane. The wear result suggested that the increasing Al particle content did not improve the wear performance of the Ni-Al composite coatings.

  9. Interfacial phenomena in the reactions of Al-B, Al-Ti-B, and Al-Zr-B alloys with KF-AlF{sub 3} and NaF-AlF{sub 3} melts

    SciTech Connect

    Lee, M.S.; Terry, B.S.; Grieveson, P.

    1993-12-01

    The interfacial phenomena occurring during the contacting of liquid Al-B, Al-Ti-B, and Al-Zr-B melts with KF-AlF{sub 3} liquid fluxes have been investigated by optical examination of quenched metal drops previously immersed in the liquid fluxes. Reactions in the Al-B/KF-AlF{sub 3} system involve the formation of metastable AlB{sub 12} at the metal/flux interface. At high KBF{sub 4} levels in the flux, the AlB{sub 12} is dispersed in the flux and also at low KBF{sub 4} levels in the metal. Reactions in the Al-Ti-B/KF-AlF{sub 3} system involve the formation of TiB{sub 2}, which may be dispersed in either the metal or the flux depending upon the composition of the flux. The results obtained for the Al-Ti-B/NaF-AlF{sub 3} and Al-Zr-B/KF-AlF{sub 3} systems were similar to those observed for the Al-Ti-B/KF-AlF{sub 3} system.

  10. Growth and Stress-induced Transformation of Zinc blende AlN Layers in Al-AlN-TiN Multilayers

    PubMed Central

    Li, Nan; Yadav, Satyesh K.; Wang, Jian; Liu, Xiang-Yang; Misra, Amit

    2015-01-01

    AlN nanolayers in sputter deposited {111}Al/AlN/TiN multilayers exhibit the metastable zinc-blende-structure (z-AlN). Based on density function theory calculations, the growth of the z-AlN is ascribed to the kinetically and energetically favored nitridation of the deposited aluminium layer. In situ nanoindentation of the as-deposited {111}Al/AlN/TiN multilayers in a high-resolution transmission electron microscope revealed the z-AlN to wurzite AlN phase transformation through collective glide of Shockley partial dislocations on every two {111} planes of the z-AlN. PMID:26681109

  11. Growth and stress-induced transformation of zinc blende AlN layers in Al-AlN-TiN multilayers

    DOE PAGES

    Li, Nan; Yadav, Satyesh K.; Wang, Jian; Liu, Xiang -Yang; Misra, Amit

    2015-12-18

    We report that AlN nanolayers in sputter deposited {111}Al/AlN/TiN multilayers exhibit the metastable zinc-blende-structure (z-AlN). Based on density function theory calculations, the growth of the z-AlN is ascribed to the kinetically and energetically favored nitridation of the deposited aluminium layer. In situ nanoindentation of the as-deposited {111}Al/AlN/TiN multilayers in a high-resolution transmission electron microscope revealed the z-AlN to wurzite AlN phase transformation through collective glide of Shockley partial dislocations on every two {111} planes of the z-AlN.

  12. Mechanical alloying of nb-al powders

    NASA Astrophysics Data System (ADS)

    Peng, Zhixue; Suryanarayana, C.; Froes, F. H. (Sam)

    1996-01-01

    The effect of mechanical alloying (MA) on solid solubility extension, nanostructure formation, amorphization, intermetallic compound formation, and the occurrence of a face-centered cubic (fcc) phase in the Nb-Al system has been studied. Solid solubility extension was observed in both the terminal compositions and intermetallic compounds: 15 pct Nb in Al and 60 pct Al in Nb, well beyond the equilibrium and even rapid solidification levels (2.4 pct Nb and 25 pct Al, respectively) and increased homogeneity range for the NbAl3 phase. Nanostructured grains formed in all compositions. In the central part of the phase diagram, amorphization occurred predominantly. Only NbAl3, the most stable intermetallic, formed during MA; in most cases, a subsequent anneal was required. On long milling time, an fcc phase, probably a nitride, formed as a result of contamination from the ambient atmosphere.

  13. Mechanical alloying of Nb-Al powders

    SciTech Connect

    Peng, Z.; Suryanarayana, C.; Froes, F.H.

    1996-01-01

    The effect of mechanical alloying (MA) on solid solubility extension, nanostructure formation, amorphization, intermetallic compound formation, and the occurrence of a face-centered cubic (fcc) phase in the Nb-Al system has been studied. Solid solubility extension was observed in both the terminal compositions and intermetallic compounds: 15 pct Nb in Al and 60 pct Al in Nb, well beyond the equilibrium and even rapid solidification levels (2.4 pct Nb and 25 pct Al, respectively) and increased homogeneity range for the NbAl{sub 3} phase. Nanostructured grains formed in all compositions. In the central part of the phase diagram, amorphization occurred predominantly. Only NbAl{sub 3}, the most stable intermetallic, formed during MA; in most cases, a subsequent anneal was required. On long milling time, an fcc phase, probably a nitride, formed as a result of contamination from the ambient atmosphere.

  14. Orientation relationship of eutectoid FeAl and FeAl2

    PubMed Central

    Scherf, A.; Kauffmann, A.; Kauffmann-Weiss, S.; Scherer, T.; Li, X.; Stein, F.; Heilmaier, M.

    2016-01-01

    Fe–Al alloys in the aluminium range of 55–65 at.% exhibit a lamellar microstructure of B2-ordered FeAl and triclinic FeAl2, which is caused by a eutectoid decomposition of the high-temperature Fe5Al8 phase, the so-called ∊ phase. The orientation relationship of FeAl and FeAl2 has previously been studied by Bastin et al. [J. Cryst. Growth (1978 ▸), 43, 745] and Hirata et al. [Philos. Mag. Lett. (2008 ▸), 88, 491]. Since both results are based on different crystallographic data regarding FeAl2, the data are re-evaluated with respect to a recent re-determination of the FeAl2 phase provided by Chumak et al. [Acta Cryst. (2010 ▸), C66, i87]. It is found that both orientation relationships match subsequent to a rotation operation of 180° about a 〈112〉 crystallographic axis of FeAl or by applying the inversion symmetry of the FeAl2 crystal structure as suggested by the Chumak data set. Experimental evidence for the validity of the previously determined orientation relationships was found in as-cast fully lamellar material (random texture) as well as directionally solidified material (∼〈110〉FeAl || solidification direction) by means of orientation imaging microscopy and global texture measurements. In addition, a preferential interface between FeAl and FeAl2 was identified by means of trace analyses using cross sectioning with a focused ion beam. On the basis of these habit planes the orientation relationship between the two phases can be described by (01)FeAl || (114) and [111]FeAl || [10]. There is no evidence for twinning within FeAl lamellae or alternating orientations of FeAl lamellae. Based on the determined orientation and interface data, an atomistic model of the structure relationship of Fe5Al8, FeAl and FeAl2 in the vicinity of the eutectoid decomposition is derived. This model is analysed with respect to the strain which has to be accommodated at the interface of FeAl and FeAl2. PMID:27047304

  15. A New Phase in ALS Research.

    PubMed

    Chong, P Andrew; Forman-Kay, Julie D

    2016-09-01

    In this issue of Structure, Conicella et al. (2016) present evidence that the low complexity C-terminal region of TDP-43 undergoes liquid-liquid phase separation. ALS-associated mutations alter this phase separation process, providing a possible mechanism for the pathology caused by these TDP-43 mutations. The work is strongly supportive of toxic loss of RNA processing function in ALS. PMID:27602988

  16. Tensile Behavior of Al2o3/feal + B and Al2o3/fecraly Composites

    NASA Technical Reports Server (NTRS)

    Draper, S. L.; Eldridge, J. I.; Aiken, B. J. M.

    1995-01-01

    The feasibility of Al2O3/FeAl + B and Al2O3/FeCrAlY composites for high-temperature applications was assessed. The major emphasis was on tensile behavior of both the monolithics and composites from 298 to 1100 K. However, the study also included determining the chemical compatibility of the composites, measuring the interfacial shear strengths, and investigating the effect of processing on the strength of the single-crystal Al2O3 fibers. The interfacial shear strengths were low for Al203/FeAl + B and moderate to high for Al203/FeCrAlY. The difference in interfacial bond strengths between the two systems affected the tensile behavior of the composites. The strength of the Al203 fiber was significantly degraded after composite processing for both composite systems and resulted in poor composite tensile properties. The ultimate tensile strength (UTS) values of the composites could generally be predicted with either rule of mixtures (ROM) calculations or existing models when using the strength of the etched-out fiber. The Al2O3/FeAl + B composite system was determined to be unfeasible due to poor interfacial shear strengths and a large mismatch in coefficient of thermal expansion (CTE). Development of the Al2O3/FeCrAlY system would require an effective diffusion barrier to minimize the fiber strength degradation during processing and elevated temperature service.

  17. Memory deficits and retrieval processes in ALS.

    PubMed

    Mantovan, M C; Baggio, L; Dalla Barba, G; Smith, P; Pegoraro, E; Soraru', G; Bonometto, P; Angelini, C

    2003-05-01

    Subtle neuropsychological deficits have been described in patients affected by amyotrophic lateral sclerosis (ALS) without dementia. Overall, selective impairment in memory function has been reported, but the source of memory impairment in ALS has yet to be defined. We performed neuropsychological screening in 20 ALS patients. Semantic encoding and post-encoding cue effects on the retrieval of word lists were investigated in the ALS patients and normal controls. Severity of memory impairment was correlated to cerebral blood perfusion detected by single photon emission computed tomography (SPECT). ALS patients showed moderate impairments in frontal and memory tests. Short-term memory was normal, while serial position retrieval of word lists with normal recency effect but poor primacy effect showed long-term memory deficit. ALS patients performed better in cued encoding than in cued post-encoding recall condition. In the cued post-encoding condition, the primacy effect in word list recall improved significantly in controls, but not in ALS patients, as compared with both the free recall and cued encoding conditions. SPECT hypoperfusion was observed in frontal and temporal areas in ALS patients. ALS patients showed a long-term memory deficit which did not improve in cued post-encoding condition as it does for controls. We hypothesize abnormal retrieval processes related to frontal lobe dysfunction which entails difficulties in generating stable long-memory traces at encoding.

  18. Memory deficits and retrieval processes in ALS.

    PubMed

    Mantovan, M C; Baggio, L; Dalla Barba, G; Smith, P; Pegoraro, E; Soraru', G; Bonometto, P; Angelini, C

    2003-05-01

    Subtle neuropsychological deficits have been described in patients affected by amyotrophic lateral sclerosis (ALS) without dementia. Overall, selective impairment in memory function has been reported, but the source of memory impairment in ALS has yet to be defined. We performed neuropsychological screening in 20 ALS patients. Semantic encoding and post-encoding cue effects on the retrieval of word lists were investigated in the ALS patients and normal controls. Severity of memory impairment was correlated to cerebral blood perfusion detected by single photon emission computed tomography (SPECT). ALS patients showed moderate impairments in frontal and memory tests. Short-term memory was normal, while serial position retrieval of word lists with normal recency effect but poor primacy effect showed long-term memory deficit. ALS patients performed better in cued encoding than in cued post-encoding recall condition. In the cued post-encoding condition, the primacy effect in word list recall improved significantly in controls, but not in ALS patients, as compared with both the free recall and cued encoding conditions. SPECT hypoperfusion was observed in frontal and temporal areas in ALS patients. ALS patients showed a long-term memory deficit which did not improve in cued post-encoding condition as it does for controls. We hypothesize abnormal retrieval processes related to frontal lobe dysfunction which entails difficulties in generating stable long-memory traces at encoding. PMID:12752394

  19. Genetics of Familial and Sporadic ALS

    ClinicalTrials.gov

    2016-03-21

    Amyotrophic Lateral Sclerosis (ALS); Familial Amyotrophic Lateral Sclerosis; Amyotrophic Lateral Sclerosis With Frontotemporal Dementia; Lou Gehrig's Disease; Motor Neuron Disease; Primary Lateral Sclerosis

  20. Superbend era begins swiftly at the ALS

    SciTech Connect

    Robinson, Art; Tamura, Lori

    2001-11-29

    The successful installation and commissioning of high-field superconducting bend magnets (superbends) in three curved sectors of ALS storage ring was the first time the magnet lattice of an operating synchrotron light source has been retrofitted in this fundamental way. As a result, the ALS now offers an expanded spectral range well into the hard x-ray region without compromising either the number of undulators or their high brightness in the soft x-ray region for which the ALS design was originally optimized. In sum, when the superbend-enhanced ALS started up for user operations in October 2001, it marked the beginning of a new era in its history.

  1. Effects of the Al content on pore structures of porous TieAl alloys

    SciTech Connect

    Jiang, Y; He, Y H; Zou, J; Huang, B; Liu, C

    2008-01-01

    PorousTi Alalloys with different nominal compositions were fabricated through a reactive synthesis of Ti and Al elemental powders. It has been found that the pore parameters vary with the Al contents, indicating that the nature of the pores can be manipulated through changing the Al contents. In addition, detailed structural characterizations showed that the fabricated porousTi Alalloys can have three crystalline phases (i.e., 2-Ti3Al, -TiAl, and TiAl3) when using different compositions. The fundamental reasons behind these phenomena have been explored.

  2. Response to Signorovitch et al.

    PubMed

    Flot, Jean-François; Debortoli, Nicolas; Hallet, Bernard; Van Doninck, Karine

    2016-08-22

    Signorovitch et al.[1] comment that an Oenothera-like meiosis [2] could produce a pattern similar to what we observed in our study of natural isolates of the bdelloid rotifer Adineta vaga, which we attributed to horizontal gene transfers (HGTs) [3]. Indeed, our HGT hypothesis appears at first sight difficult to conciliate with their observation of a congruent pattern of allele sharing at four large loci possibly located on different chromosomes [4]. However, one might imagine conditions under which massive horizontal gene transfer between bdelloid individuals could produce such a pattern, notably if the individuals involved had previously lost most of their heterozygosity because of their exposure to frequent desiccation (which produces DNA double-strand breaks [5]). In the published A. vaga genome the loss of heterozygosity due to large-scale gene conversion events or break-induced replication covers only about 10% of the genome [6], but this percentage may be much higher in environmental isolates that often experience dessication. Besides, if an Oenothera-like mode of meiosis occurs in bdelloids frequently enough to be detected in a single sampling of 29 individuals (as in [4]), one would expect males and meiosis to be observed at least occasionally, and instances of congruent allele sharing across loci should turn up frequently in genetic surveys. This was not the case in [3]: among the 82 A. vaga individuals sequenced for four nuclear markers, no trio of individuals presented congruent patterns of shared sequences at different loci. For these reasons, and in the absence of any direct evidence for an Oenothera-like meiosis in bdelloids, we still consider inter-bdelloid HGTs a more parsimonious explanation for our results. PMID:27554651

  3. Atomic structure and oxygen deficiency of the ultrathin aluminium oxide barrier in Al/AlOx/Al Josephson junctions

    NASA Astrophysics Data System (ADS)

    Zeng, Lunjie; Tran, Dung Trung; Tai, Cheuk-Wai; Svensson, Gunnar; Olsson, Eva

    2016-07-01

    Al/AlOx/Al Josephson junctions are the building blocks of a wide range of superconducting quantum devices that are key elements for quantum computers, extremely sensitive magnetometers and radiation detectors. The properties of the junctions and the superconducting quantum devices are determined by the atomic structure of the tunnel barrier. The nanoscale dimension and disordered nature of the barrier oxide have been challenges for the direct experimental investigation of the atomic structure of the tunnel barrier. Here we show that the miniaturized dimension of the barrier and the interfacial interaction between crystalline Al and amorphous AlOx give rise to oxygen deficiency at the metal/oxide interfaces. In the interior of the barrier, the oxide resembles the atomic structure of bulk aluminium oxide. Atomic defects such as oxygen vacancies at the interfaces can be the origin of the two-level systems and contribute to decoherence and noise in superconducting quantum circuits.

  4. Strengthening of Al and Al-Mg alloy wires by melt inoculation with Al/MgB2 nanocomposite

    NASA Astrophysics Data System (ADS)

    Florián-Algarín, David; Marrero, Raúl; Padilla, Alexandra; Suárez, Oscar Marcelo

    2015-12-01

    This study hinges on the feasibility of strengthening Al and Al-Mg wires by adding Al nanocomposite pellets containing MgB2 nanoparticles into the melt upon fabrication. These MgB2 nanoparticles were obtained by fragmentation using a high-energy ball mill, and were, afterward, mechanically alloyed with pure aluminum. The resulting MgB2/Al nanocomposite pellets were sintered at 260°C to be subsequently added into molten aluminum and an Al-Mg alloy melt. Cold rolling intercalated with stepwise annealing allowed the fabrication of 1 mm diameter wires with a final area reduction of 96%. Mechanical and physical properties of the treated wire specimens were compared to those of similarly processed pure aluminum wire. The ultimate tensile strength of the treated wires increased approximately double fold with respect to untreated wires at the expense of some loss in electrical conductivity.

  5. Atomic structure and oxygen deficiency of the ultrathin aluminium oxide barrier in Al/AlOx/Al Josephson junctions.

    PubMed

    Zeng, Lunjie; Tran, Dung Trung; Tai, Cheuk-Wai; Svensson, Gunnar; Olsson, Eva

    2016-01-01

    Al/AlOx/Al Josephson junctions are the building blocks of a wide range of superconducting quantum devices that are key elements for quantum computers, extremely sensitive magnetometers and radiation detectors. The properties of the junctions and the superconducting quantum devices are determined by the atomic structure of the tunnel barrier. The nanoscale dimension and disordered nature of the barrier oxide have been challenges for the direct experimental investigation of the atomic structure of the tunnel barrier. Here we show that the miniaturized dimension of the barrier and the interfacial interaction between crystalline Al and amorphous AlOx give rise to oxygen deficiency at the metal/oxide interfaces. In the interior of the barrier, the oxide resembles the atomic structure of bulk aluminium oxide. Atomic defects such as oxygen vacancies at the interfaces can be the origin of the two-level systems and contribute to decoherence and noise in superconducting quantum circuits. PMID:27403611

  6. Atomic structure and oxygen deficiency of the ultrathin aluminium oxide barrier in Al/AlOx/Al Josephson junctions

    PubMed Central

    Zeng, Lunjie; Tran, Dung Trung; Tai, Cheuk-Wai; Svensson, Gunnar; Olsson, Eva

    2016-01-01

    Al/AlOx/Al Josephson junctions are the building blocks of a wide range of superconducting quantum devices that are key elements for quantum computers, extremely sensitive magnetometers and radiation detectors. The properties of the junctions and the superconducting quantum devices are determined by the atomic structure of the tunnel barrier. The nanoscale dimension and disordered nature of the barrier oxide have been challenges for the direct experimental investigation of the atomic structure of the tunnel barrier. Here we show that the miniaturized dimension of the barrier and the interfacial interaction between crystalline Al and amorphous AlOx give rise to oxygen deficiency at the metal/oxide interfaces. In the interior of the barrier, the oxide resembles the atomic structure of bulk aluminium oxide. Atomic defects such as oxygen vacancies at the interfaces can be the origin of the two-level systems and contribute to decoherence and noise in superconducting quantum circuits. PMID:27403611

  7. Atomic structure and oxygen deficiency of the ultrathin aluminium oxide barrier in Al/AlOx/Al Josephson junctions.

    PubMed

    Zeng, Lunjie; Tran, Dung Trung; Tai, Cheuk-Wai; Svensson, Gunnar; Olsson, Eva

    2016-07-12

    Al/AlOx/Al Josephson junctions are the building blocks of a wide range of superconducting quantum devices that are key elements for quantum computers, extremely sensitive magnetometers and radiation detectors. The properties of the junctions and the superconducting quantum devices are determined by the atomic structure of the tunnel barrier. The nanoscale dimension and disordered nature of the barrier oxide have been challenges for the direct experimental investigation of the atomic structure of the tunnel barrier. Here we show that the miniaturized dimension of the barrier and the interfacial interaction between crystalline Al and amorphous AlOx give rise to oxygen deficiency at the metal/oxide interfaces. In the interior of the barrier, the oxide resembles the atomic structure of bulk aluminium oxide. Atomic defects such as oxygen vacancies at the interfaces can be the origin of the two-level systems and contribute to decoherence and noise in superconducting quantum circuits.

  8. Influence of AlN thickness on AlGaN epilayer grown by MOCVD

    NASA Astrophysics Data System (ADS)

    Jayasakthi, M.; Juillaguet, S.; Peyre, H.; Konczewicz, L.; Baskar, K.; Contreras, S.

    2016-10-01

    AlGaN/AlN layers were grown by metalorganic chemical vapor deposition (MOCVD) on sapphire substrates. The AlN buffer thickness was varied from 400 nm to 800 nm. The AlGaN layer thickness was 1000 nm. The crystalline quality, thickness and composition of AlGaN were determined using high resolution X-ray diffraction (HRXRD). The threading dislocation density (TDD) was found to decrease with increase of AlN layer thickness. Reciprocal space mapping (RSM) was used to estimate the strain and relaxation between AlGaN and AlN. The optical properties of AlGaN layers were investigated by temperature dependent photoluminescence (PL). PL intensities of AlGaN layers increases with increasing the AlN thickness. The surface morphology of AlGaN was studied by atomic force microscopy (AFM). Root mean square (RMS) roughness values were found to be decreased while increase of AlN thickness.

  9. Diana Al-Hadid: Identity and Heritage

    ERIC Educational Resources Information Center

    Jungerberg, Tom; Smith, Anna; Borsh, Colleen

    2012-01-01

    Diana Al-Hadid's sculptures reflect the many locations, cultures, histories, and mythologies that have shaped her as an artist. In large-scale works which have the appearance of architectural ruins, Al-Hadid employs imagery drawn from many diverse interests including science and technology, history, and literature. She also incorporates images and…

  10. Quantitatively Probing the Al Distribution in Zeolites

    SciTech Connect

    Vjunov, Aleksei; Fulton, John L.; Huthwelker, Thomas; Pin, Sonia; Mei, Donghai; Schenter, Gregory K.; Govind, Niranjan; Camaioni, Donald M.; Hu, Jian Z.; Lercher, Johannes A.

    2014-06-11

    The degree of substitution of Si4+ by Al3+ in the oxygen-terminated tetrahedra (Al T-sites) of zeolites determines the concentration of ion-exchange and Brønsted acid sites. As the location of the tetrahedra and the associated subtle variations in bond angles influence the acid strength, quantitative information about Al T-sites in the framework is critical to rationalize catalytic properties and to design new catalysts. A quantitative analysis is reported that uses a combination of extended X-ray absorption fine structure (EXAFS) analysis and 27Al MAS NMR spectroscopy supported by DFT-based molecular dynamics simulations. To discriminate individual Al atoms, sets of ab initio EXAFS spectra for various T-sites are generated from DFT-based molecular dynamics simulations allowing quantitative treatment of the EXAFS single- and multiple-photoelectron scattering processes out to 3-4 atom shells surrounding the Al absorption center. It is observed that identical zeolite types show dramatically different Al-distributions. A preference of Al for T-sites that are part of one or more 4-member rings in the framework over those T-sites that are part of only 5- and 6-member rings in the HBEA150 sample has been determined from a combination of these methods. This work was supported by the U. S. Department of Energy (DOE), Office of Basic Energy Sciences, Division of Chemical Sciences, Geosciences & Biosciences.

  11. Durability Assessment of TiAl Alloys

    NASA Technical Reports Server (NTRS)

    Draper, Susan L.; Lerch, Bradley A.

    2008-01-01

    The durability of TiAl is a prime concern for the implementation of TiAl into aerospace engines. Two durability issues, the effect of high temperature exposure on mechanical properties and impact resistance, have been investigated and the results are summarized in this paper. Exposure to elevated temperatures has been shown to be detrimental to the room temperature ductility of gamma alloys with the most likely mechanisms being the ingress of interstitials from the surface. Fluorine ion implantation has been shown to improve the oxidation resistance of gamma alloys, and ideally it could also improve the environmental embrittlement of high Nb content TiAl alloys. The effect of F ion implantation on the surface oxidation and embrittlement of a third generation, high Nb content TiAl alloy (Ti-45Al-5Nb-B-C) were investigated. Additionally, the ballistic impact resistance of a variety of gamma alloys, including Ti-48Al-2Cr- 2Nb, Ti-47Al-2Cr-2Nb, ABB-2, ABB-23, NCG359E, 95A and Ti-45Al-5Nb-B-C was accessed. Differences in the ballistic impact properties of the various alloys will be discussed, particularly with respect to their manufacturing process, microstructure, and tensile properties.

  12. Energetics of Al13 Keggin cluster compounds

    PubMed Central

    Armstrong, Christopher R.; Casey, William H.; Navrotsky, Alexandra

    2011-01-01

    The ϵ-Al13 Keggin aluminum hydroxide clusters are essential models in establishing molecular pathways for geochemical reactions. Enthalpies of formation are reported for two salts of aluminum centered ϵ-Keggin clusters, Al13 selenate, (Na(AlO4)Al12(OH)24(SeO4)4•12H2O) and Al13 sulfate, (NaAlO4Al12(OH)24(SO4)4•12H2O). The measured enthalpies of solution, ΔHsol, at 28 °C in 5 N HCl for the ε-Al13 selenate and sulfate are −924.57 (± 3.83) and −944.30 ( ± 5.66) kJ·mol-1, respectively. The enthalpies of formation from the elements, ΔHf,el, for Al13 selenate and sulfate are −19,656.35 ( ± 67.30) kJ·mol-1, and −20,892.39 ( ± 70.01) kJ·mol-1, respectively. In addition, ΔHf,el for sodium selenate decahydrate was calculated using data from high temperature oxide melt solution calorimetry measurements: −4,006.39 ( ± 11.91) kJ·mol-1. The formation of both ε-Al13 Keggin cluster compounds is exothermic from oxide-based components but energetically unfavorable with respect to a gibbsite-based assemblage. To understand the relative affinity of the ϵ-Keggin clusters for selenate and sulfate, the enthalpy associated with two S-Se exchange reactions was calculated. In the solid state, selenium is favored in the Al13 compound relative to the binary chalcogenate, while in 5 N HCl, sulfur is energetically favored in the cluster compound compared to the aqueous solution. This contribution represents the first thermodynamic study of ε-Al13 cluster compounds and establishes a method for other such molecules, including the substituted versions that have been created for kinetic studies. Underscoring the importance of ε-Al13 clusters in natural and anthropogenic systems, these data provide conclusive thermodynamic evidence that the Al13 Keggin cluster is a crucial intermediate species in the formation pathway from aqueous aluminum monomers to aluminum hydroxide precipitates. PMID:21852572

  13. NiAl alloys for structural uses

    NASA Technical Reports Server (NTRS)

    Koss, D. A.

    1991-01-01

    Alloys based on the intermetallic compound NiAl are of technological interest as high temperature structural alloys. These alloys possess a relatively low density, high melting temperature, good thermal conductivity, and (usually) good oxidation resistance. However, NiAl and NiAl-base alloys suffer from poor fracture resistance at low temperatures as well as inadequate creep strength at elevated temperatures. This research program explored macroalloying additions to NiAl-base alloys in order to identify possible alloying and processing routes which promote both low temperature fracture toughness and high temperature strength. Initial results from the study examined the additions of Fe, Co, and Hf on the microstructure, deformation, and fracture resistance of NiAl-based alloys. Of significance were the observations that the presence of the gamma-prime phase, based on Ni3Al, could enhance the fracture resistance if the gamma-prime were present as a continuous grain boundary film or 'necklace'; and the Ni-35Al-20Fe alloy was ductile in ribbon form despite a microstructure consisting solely of the B2 beta phase based on NiAl. The ductility inherent in the Ni-35Al-20Fe alloy was explored further in subsequent studies. Those results confirm the presence of ductility in the Ni-35Al-20Fe alloy after rapid cooling from 750 - 1000 C. However exposure at 550 C caused embrittlement; this was associated with an age-hardening reaction caused by the formation of Fe-rich precipitates. In contrast, to the Ni-35Al-20Fe alloy, exploratory research indicated that compositions in the range of Ni-35Al-12Fe retain the ordered B2 structure of NiAl, are ductile, and do not age-harden or embrittle after thermal exposure. Thus, our recent efforts have focused on the behavior of the Ni-35Al-12Fe alloy. A second parallel effort initiated in this program was to use an alternate processing technique, mechanical alloying, to improve the properties of NiAl-alloys. Mechanical alloying in the

  14. Corrosion Studies of 2195 Al-Li Alloy and 2219 Al Alloy with Differing Surface Treatments

    NASA Technical Reports Server (NTRS)

    Danford, M. D.; Mendrek, M. J.

    1998-01-01

    Corrosion studies of 2195 Al-Li and 2219 Al alloys have been conducted using the scanning reference electrode technique (SRET) and the polarization resistance (PR) technique. The SRET was used to study corrosion mechanisms, while corrosion rate measurements were studied with the PR technique. Plates of Al203 blasted, soda blasted and conversion coated 2219 Al were coated with Deft primer and the corrosion rates studied with the EIS technique. Results from all of these studies are presented.

  15. Consolidation of Al2O3/Al Nanocomposite Powder by Cold Spray

    NASA Astrophysics Data System (ADS)

    Poirier, Dominique; Legoux, Jean-Gabriel; Drew, Robin A. L.; Gauvin, Raynald

    2011-01-01

    While the improvement in mechanical properties of nanocomposites makes them attractive materials for structural applications, their processing still presents significant challenges. In this article, cold spray was used to consolidate milled Al and Al2O3/Al nanocomposite powders as well as the initial unmilled and unreinforced Al powder. The microstructure and nanohardness of the feedstock powders as well as those of the resulting coatings were compared. The results show that the large increase in hardness of the Al powder after mechanical milling is preserved after cold spraying. Good quality coating with low porosity is obtained from milled Al. However, the addition of Al2O3 to the Al powder during milling decreases the powder and coating nanohardness. This lower hardness is attributed to non-optimized milling parameters leading to cracked particles with insufficient Al2O3 embedding in Al. The coating produced from the milled Al2O3/Al mixture also showed lower particle cohesion and higher amount of porosity.

  16. Japanese version of the ALS-FTD-Questionnaire (ALS-FTD-Q-J).

    PubMed

    Watanabe, Yasuhiro; Beeldman, Emma; Raaphorst, Joost; Izumi, Yuishin; Yoshino, Hiide; Masuda, Michihito; Atsuta, Naoki; Ito, Satoru; Adachi, Tadashi; Adachi, Yoshiki; Yokota, Osamu; Oda, Masaya; Hanashima, Ritsuko; Ogino, Mieko; Ichikawa, Hiroo; Hasegawa, Kazuko; Kimura, Hideki; Shimizu, Toshio; Aiba, Ikuko; Yabe, Hayato; Kanba, Makoto; Kusumi, Kimiyoshi; Aoki, Tetsuya; Hiroe, Yu; Watanabe, Hirohisa; Nishiyama, Kazutoshi; Nomoto, Masahiro; Sobue, Gen; Nakashima, Kenji

    2016-08-15

    Amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD) share common clinical, genetic and neuropathological features. Some ALS patients have behavioral/personality changes, which could result in significant obstacles in the care provided by family members and caregivers. An easy screening tool would contribute greatly to the evaluation of these symptoms. We translated the ALS-FTD-Questionnaire, developed in the Netherlands, into Japanese (ALS-FTD-Q-J) and examined the clinimetric properties (internal consistency, construct and clinical validity). Patients with ALS and/or behavioral variant FTD (bvFTD) were evaluated alongside healthy controls in this multicenter study. All ALS patients, regardless of bvFTD status, were further evaluated by the frontal behavioral inventory (FBI) and for frontal/executive function, cognition, anxiety/depression, and motor functions. Data from 146 subjects were analyzed: ALS (92), ALS-bvFTD (6), bvFTD (16), and healthy controls (32). The internal consistency of the ALS-FTD-Q-J was good (Cronbach α=0.92). The ALS-FTD-Q-J showed construct validity as it exhibited a high correlation with the FBI (r=0.79). However, correlations were moderate with anxiety/depression and low with cognitive scales, in contrast to the original report, i.e. a moderate correlation with cognition and a low correlation with anxiety/depression. The ALS-FTD-Q-J discriminated ALS patients from (ALS-)bvFTD patients and controls. Thus, the ALS-FTD-Q-J is useful for evaluating Japanese ALS/FTD patients.

  17. Japanese version of the ALS-FTD-Questionnaire (ALS-FTD-Q-J).

    PubMed

    Watanabe, Yasuhiro; Beeldman, Emma; Raaphorst, Joost; Izumi, Yuishin; Yoshino, Hiide; Masuda, Michihito; Atsuta, Naoki; Ito, Satoru; Adachi, Tadashi; Adachi, Yoshiki; Yokota, Osamu; Oda, Masaya; Hanashima, Ritsuko; Ogino, Mieko; Ichikawa, Hiroo; Hasegawa, Kazuko; Kimura, Hideki; Shimizu, Toshio; Aiba, Ikuko; Yabe, Hayato; Kanba, Makoto; Kusumi, Kimiyoshi; Aoki, Tetsuya; Hiroe, Yu; Watanabe, Hirohisa; Nishiyama, Kazutoshi; Nomoto, Masahiro; Sobue, Gen; Nakashima, Kenji

    2016-08-15

    Amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD) share common clinical, genetic and neuropathological features. Some ALS patients have behavioral/personality changes, which could result in significant obstacles in the care provided by family members and caregivers. An easy screening tool would contribute greatly to the evaluation of these symptoms. We translated the ALS-FTD-Questionnaire, developed in the Netherlands, into Japanese (ALS-FTD-Q-J) and examined the clinimetric properties (internal consistency, construct and clinical validity). Patients with ALS and/or behavioral variant FTD (bvFTD) were evaluated alongside healthy controls in this multicenter study. All ALS patients, regardless of bvFTD status, were further evaluated by the frontal behavioral inventory (FBI) and for frontal/executive function, cognition, anxiety/depression, and motor functions. Data from 146 subjects were analyzed: ALS (92), ALS-bvFTD (6), bvFTD (16), and healthy controls (32). The internal consistency of the ALS-FTD-Q-J was good (Cronbach α=0.92). The ALS-FTD-Q-J showed construct validity as it exhibited a high correlation with the FBI (r=0.79). However, correlations were moderate with anxiety/depression and low with cognitive scales, in contrast to the original report, i.e. a moderate correlation with cognition and a low correlation with anxiety/depression. The ALS-FTD-Q-J discriminated ALS patients from (ALS-)bvFTD patients and controls. Thus, the ALS-FTD-Q-J is useful for evaluating Japanese ALS/FTD patients. PMID:27423564

  18. A new discontinuously reinforced aluminum MMC: Al+AlB{sub 2} flakes

    SciTech Connect

    HALL,AARON C.; ECONOMY,J.

    2000-06-08

    Development of a novel metal matrix composite based on the Al-B alloy system has been undertaken. Preparation of this discontinuously reinforced material is based on the precipitation of high aspect ratio AlB{sub 2} from an Al-B alloy. This paper describes a number of efforts forced on preparing high volume fractions (> 30 v%) of AlB{sub 2} in aluminum. New insights into the behavior of the Al-B alloys system allowed this effort to be successful.

  19. Atomistic Modeling of RuAl and (RuNi) Al Alloys

    NASA Technical Reports Server (NTRS)

    Gargano, Pablo; Mosca, Hugo; Bozzolo, Guillermo; Noebe, Ronald D.; Gray, Hugh R. (Technical Monitor)

    2002-01-01

    Atomistic modeling of RuAl and RuAlNi alloys, using the BFS (Bozzolo-Ferrante-Smith) method for alloys is performed. The lattice parameter and energy of formation of B2 RuAl as a function of stoichiometry and the lattice parameter of (Ru(sub 50-x)Ni(sub x)Al(sub 50)) alloys as a function of Ni concentration are computed. BFS based Monte Carlo simulations indicate that compositions close to Ru25Ni25Al50 are single phase with no obvious evidence of a miscibility gap and separation of the individual B2 phases.

  20. Effects of hydrogen absorption in TbNiAl and UNiAl

    SciTech Connect

    Bordallo, H.N.; Nakotte, H.; Schultz, A.; Kolomiets, A.V.; Havela, L.; Andreev, A.V.

    1998-12-31

    Although hydrides of intermetallic compounds are used extensively as hydrogen-storage media, little is known about the exact nature of metal-hydrogen interactions. However, this knowledge is of essential importance for the understanding of thermodynamics and other properties. Hydrides (deuterides) of TbNiAl and UNiAl have been widely studied because of drastic increase of magnetic ordering temperature under hydrogenation. Here the authors report neutron-diffraction results of the three deuterides, TbNiAlD{sub 1.28}, TbNiAlD{sub 0.8}a nd UNiAlD{sub 2.23}.

  1. Arsenate uptake by Al nanoclusters and other Al-based sorbents during water treatment.

    PubMed

    Mertens, Jasmin; Rose, Jérôme; Wehrli, Bernhard; Furrer, Gerhard

    2016-01-01

    In many parts of the world, arsenic from geogenic and anthropogenic sources deteriorates the quality of drinking water resources. Effective methods of arsenic removal include adsorption and coagulation with iron- and aluminum-based materials, of which polyaluminum chloride is widely employed as coagulant in water treatment due to its low cost and high efficiency. We compared the arsenic uptake capacity and the arsenic bonding sites of different Al-based sorbents, including Al nanoclusters, polyaluminum chloride, polyaluminum granulate, and gibbsite. Extended X-ray absorption fine structure (EXAFS) spectroscopy revealed that As(V) forms bidentate-binuclear complexes in interaction with all Al-based removal agents. The octahedral configuration of nanoclusters and the distribution of sorption sites remain the same in all types of removal agents consisting of nano-scale Al oxyhydroxide particles. The obtained distances for As(V)-O and As(V)-Al agreed with previously published data and were found to be 1.69 ± 0.02 Å and 3.17-3.21 Å, respectively. Our study suggests that As(V) binds to Al nanoclusters as strongly as to Al oxide surfaces. The As sorption capacity of Al nanoclusters was found to be very similar to that of Al clusters in a polyaluminum chloride. The most efficient Al-based sorbents for arsenic removal were Al nanoclusters, followed by polyaluminum granulate. PMID:26613179

  2. Hybrid Al + Al3Ni metallic foams synthesized in situ via laser engineered net shaping

    NASA Astrophysics Data System (ADS)

    Zheng, Baolong; Li, Ying; Smugeresky, John E.; Zhou, Yizhang; Baker, Dean; Lavernia, Enrique J.

    2011-09-01

    A hybrid, Al + Al3Ni metallic foam was synthesized in situ via laser engineered net shaping (LENS®) of Ni-coated 6061 Al powder in the absence of a foaming agent. During LENS® processing, the Ni coating reacted with the Al matrix, resulting in the simultaneous formation of a fine dispersion of Al3Ni, and a high volume fraction of porosity. As a reinforcement phase, the intermetallic compound formed particles with a size range of 1-5 µm and a volume fraction of 63%, with accompanying 35-300 µm pores with a 60% volume fraction. The microstructure of the as-deposited Al + Al3Ni composite foams was characterized using SEM, EDS, XRD and TEM/HRTEM techniques. The evolution of the microstructure was analyzed on the basis of the thermal field present during deposition, paying particular attention to the thermodynamics of the Al3Ni intermetallic compound formation as well as discussing the mechanisms that may be responsible for the observed porosity. The mechanical behavior of the as-deposited material was characterized using compression and microhardness testing, indicating that the yield strength and hardness are 190 MPa and 320 HV, respectively, which represents an increase of over three times higher than that of annealed Al6061, or similar to heat-treated Al6061 fully dense matrix, and much higher than those of traditional Al alloy foams, and with a low density of 1.64 g/m3.

  3. Ni{sub 3}Al aluminide alloys

    SciTech Connect

    Liu, C.T.

    1993-10-01

    This paper provides a brief review of the recent progress in research and development of Ni{sub 3}Al and its alloys. Emphasis has been placed on understanding low ductility and brittle fracture of Ni{sub 3}Al alloys at ambient and elevated temperatures. Recent studies have resulted in identifying both intrinsic and extrinsic factors governing the fracture behavior of Ni{sub 3}Al alloys. Parallel efforts on alloy design using physical metallurgy principles have led to properties for structural use. Industrial interest in these alloys is high, and examples of industrial involvement in processing and utilization of these alloys are briefly mentioned.

  4. InAlN/AlN/GaN heterostructures for high electron mobility transistors

    NASA Astrophysics Data System (ADS)

    Usov, S. O.; Sakharov, A. V.; Tsatsulnikov, A. F.; Lundin, V. W.; Zavarin, E. E.; Nikolaev, A. E.; Yagovkina, M. A.; Zemlyakov, V. E.; Egorkin, V. I.; Ustinov, V. M.

    2016-08-01

    The results of development of InAlN/AlN/GaN heterostructures, grown on sapphire substrates by metal-organic chemical vapour deposition, and high electron mobility transistors (HEMTs) based on them are presented. The dependencies of the InAlN/AlN/GaN heterostructure properties on epitaxial growth conditions were investigated. The optimal indium content and InAlN barrier layer thicknesses of the heterostructures for HEMT s were determined. The possibility to improve the characteristics of HEMTs by in-situ passivation by Si3N4 thin protective layer deposited in the same epitaxial process was demonstrated. The InAlN/AlN/GaN heterostructure grown on sapphire substrate with diameter of 100 mm were obtained with sufficiently uniform distribution of sheet resistance. The HEMTs with saturation current of 1600 mA/mm and transconductance of 230 mS/mm are demonstrated.

  5. A Rechargeable Al/S Battery with an Ionic-Liquid Electrolyte.

    PubMed

    Gao, Tao; Li, Xiaogang; Wang, Xiwen; Hu, Junkai; Han, Fudong; Fan, Xiulin; Suo, Liumin; Pearse, Alex J; Lee, Sang Bok; Rubloff, Gary W; Gaskell, Karen J; Noked, Malachi; Wang, Chunsheng

    2016-08-16

    Aluminum metal is a promising anode material for next generation rechargeable batteries owing to its abundance, potentially dendrite-free deposition, and high capacity. The rechargeable aluminum/sulfur (Al/S) battery is of great interest owing to its high energy density (1340 Wh kg(-1) ) and low cost. However, Al/S chemistry suffers poor reversibility owing to the difficulty of oxidizing AlSx . Herein, we demonstrate the first reversible Al/S battery in ionic-liquid electrolyte with an activated carbon cloth/sulfur composite cathode. Electrochemical, spectroscopic, and microscopic results suggest that sulfur undergoes a solid-state conversion reaction in the electrolyte. Kinetics analysis identifies that the slow solid-state sulfur conversion reaction causes large voltage hysteresis and limits the energy efficiency of the system.

  6. High performance AlGaN/GaN HEMTs with AlN/SiNx passivation

    NASA Astrophysics Data System (ADS)

    Xin, Tan; Yuanjie, Lü; Guodong, Gu; Li, Wang; Shaobo, Dun; Xubo, Song; Hongyu, Guo; Jiayun, Yin; Shujun, Cai; Zhihong, Feng

    2015-07-01

    AlGaN/GaN high electron-mobility transistors (HEMTs) with 5 nm AlN passivation by plasma enhanced atomic layer deposition (PEALD) were fabricated, covered by 50 nm SiNx which was grown by plasma enhanced chemical vapor deposition (PECVD). With PEALD AlN passivation, current collapse was suppressed more effectively and the devices show better subthreshold characteristics. Moreover, the insertion of AlN increased the RF transconductance, which lead to a higher cut-off frequency. Temperature dependence of DC characteristics demonstrated that the degradations of drain current and maximum transconductance at elevated temperatures for the AlN/SiNx passivated devices were much smaller compared with the devices with SiNx passivation, indicating that PEALD AlN passivation can improve the high temperature operation of the AlGaN/GaN HEMTs. Project supported by the National Natural Science Foundation of China (No. 60890192).

  7. A Rechargeable Al/S Battery with an Ionic-Liquid Electrolyte.

    PubMed

    Gao, Tao; Li, Xiaogang; Wang, Xiwen; Hu, Junkai; Han, Fudong; Fan, Xiulin; Suo, Liumin; Pearse, Alex J; Lee, Sang Bok; Rubloff, Gary W; Gaskell, Karen J; Noked, Malachi; Wang, Chunsheng

    2016-08-16

    Aluminum metal is a promising anode material for next generation rechargeable batteries owing to its abundance, potentially dendrite-free deposition, and high capacity. The rechargeable aluminum/sulfur (Al/S) battery is of great interest owing to its high energy density (1340 Wh kg(-1) ) and low cost. However, Al/S chemistry suffers poor reversibility owing to the difficulty of oxidizing AlSx . Herein, we demonstrate the first reversible Al/S battery in ionic-liquid electrolyte with an activated carbon cloth/sulfur composite cathode. Electrochemical, spectroscopic, and microscopic results suggest that sulfur undergoes a solid-state conversion reaction in the electrolyte. Kinetics analysis identifies that the slow solid-state sulfur conversion reaction causes large voltage hysteresis and limits the energy efficiency of the system. PMID:27417442

  8. Fabrication and characterization of all-refractory NbCN/Al/AlO(x)/Al/Nb junctions

    NASA Astrophysics Data System (ADS)

    Barber, Z. H.; Blamire, M. G.; Somekh, R. E.; Evetts, J. E.

    1993-03-01

    High-quality AlO(x) tunnel barriers have been fabricated on epitaxial niobium carbonitride (NbCN) base layers by the deposition of an Al layer followed by thermal oxidation. By careful control of its uniformity, the thickness of the Al layer has been reduced to less than 3 nm, which results in an average gap voltage, Vg(NbCN), of up to 2.65 mV. Using a self-aligned whole-wafer processing route, high-quality NbCN/Al/AlO(x)/Al/Nb junctions as small as 0.6 sq mm have been made. These junctions offer considerable advantages over directly deposited barriers in terms of minimal subgap leakage, good control of the barrier conductance, and simple processing procedures. It was shown that submicron junctions can be fabricated with no gap smearing or reduction in quality. Using only Nb counterelectrodes total gap voltages up to 4.0 mV, with widths of 0.6 mV, have been demonstrated in high-quality junctions.

  9. Characterization of AlInN/AlN/GaN Heterostructures with Different AlN Buffer Thickness

    NASA Astrophysics Data System (ADS)

    Çörekçi, S.; Dugan, S.; Öztürk, M. K.; Çetin, S. Ş.; Çakmak, M.; Özçelik, S.; Özbay, E.

    2016-07-01

    Two AlInN/AlN/GaN heterostructures with 280-nm- and 400-nm-thick AlN buffer grown on sapphire substrates by metal-organic chemical vapor deposition (MOCVD) have been investigated by x-ray diffraction (XRD), atomic force microscopy (AFM), photoluminescence (PL) and Hall-effect measurements. The symmetric (0002) plane with respect to the asymmetric (10bar{1}2) plane in the 280-nm-thick AlN buffer has a higher crystal quality, as opposed to the 400-nm-thick buffer. The thinner buffer improves the crystallinity of both (0002) and (10bar{1}2) planes in the GaN layers, it also provides a sizeable reduction in dislocation density of GaN. Furthermore, the lower buffer thickness leads to a good quality surface with an rms roughness of 0.30 nm and a dark spot density of 4.0 × 108 cm-2. The optical and transport properties of the AlInN/AlN/GaN structure with the relatively thin buffer are compatible with the enhancement in its structural quality, as verified by XRD and AFM results.

  10. Interdiffusion in Diffusion Couples: U-Mo v. Al and Al-Si

    SciTech Connect

    D. D. Keiser, Jr.; E. Perez; B. Yao; Y. H. Sohn

    2009-11-01

    Interdiffusion and microstructural development in the U-Mo-Al system was examined using solid-tosolid diffusion couples consisting of U-7wt.%Mo, U-10wt.%Mo and U-12wt.%Mo vs. pure Al, annealed at 600°C for 24 hours. The influence of Si alloying addition (up to 5 wt.%) in Al on the interdiffusion microstructural development was also examined using solid-to-solid diffusion couples consisting of U-7wt.%Mo, U-10wt.%Mo and U-12wt.%Mo vs. pure Al, Al-2wt.%Si, and Al-5wt.%Si annealed at 550°C up to 20 hours. Scanning electron microscopy (SEM), transmission electron microscopy (TEM) and electron probe microanalysis (EPMA) were employed to examine the development of a very fine multiphase intermetallic layer. In ternary U-Mo-Al diffusion couples annealed at 600°C for 24 hours, interdiffusion microstructure varied of finely dispersed UAl3, UAl4, U6Mo4Al43, and UMo2Al20 phases while the average composition throughout the interdiffusion zone remained constant at approximately 80 at.% Al. Interdiffusion microstructure observed by SEM/TEM analyses and diffusion paths drawn from concentration profiles determined by EPMA appear to deviate from the assumption of “local thermodynamic equilibrium,” and suggest that interdiffusion occurs via supersaturated UAl4 followed by equilibrium transformation into UAl3, U6Mo4Al43, UAl4 and UMo2Al20 phases. Similar observation was made for U-Mo vs. Al diffusion couples annealed at 550°C. The addition of Si (up to 5 wt.%) in Al significantly reduced the thickness of the intermetallic layer by changing the constituent phases of the interdiffusion zone developed in U-Mo vs. Al-Si diffusion couples. Specifically, the formation of (U,Mo)(Al,Si)3 with relatively large solubility for Mo and Si, along with UMo2Al20 phases was observed along with disappearance of U6Mo4Al43 and UAl4 phases. Simplified understanding based on U-Al, U-Si, and Mo-Si binary phase diagrams is discussed in the light of the beneficial effect of Si alloying addition.

  11. Smoking Tied to Shorter Survival with ALS

    MedlinePlus

    ... with ALS between 2007 and 2011 in northern Italy. They also looked at chronic lung disease (COPD) ... of neuroscience at the University of Turin in Italy, led the study. SOURCE: BMJ , news release, Sept. ...

  12. Stem Cells Deemed Safe for ALS Patients

    MedlinePlus

    ... baby step forward," said lead researcher Dr. Jonathan Glass, of Emory University, in Atlanta. "We can say this procedure is doable in ALS patients," Glass said. "Now we have to test whether it's ...

  13. Glial cells in ALS: the missing link?

    PubMed

    Raibon, Elsa; Todd, Lisa Marie; Möller, Thomas

    2008-08-01

    Amyotrophic lateral sclerosis (ALS) was initially known as Charcot's sclerosis, named after the French neurobiologist and physician Jean-Martin Charcot who first described this type of muscular atrophy in the early nineteenth century. In the United States, ALS became widely known as Lou Gehrig's disease after the famous baseball player who succumbed to the disease in the late 1930s. Currently, ALS is the most common motor neuron disease, with a worldwide incidence of 8 cases per 100,000 population per year. Familial forms constitute approximately 5% to 10% of all cases. Onset increases with age, with a peak in the seventh decade and a slight preponderance (relative risk, 1.3-1.5) among men compared with women. Rapid progression of motor neuron loss leads to death an average of 3 to 5 years after symptom onset. The cause of ALS remains unknown and there is still no curative therapy.

  14. Thermal Conductivity of Al-Salt Composites

    NASA Astrophysics Data System (ADS)

    Li, Peng; Zhang, Mei; Wang, Lijun; Seetharaman, Seshadri

    2015-11-01

    With a view to examine the possibility of estimating the content of entrapped metallic aluminium in the salt cake from aluminium remelting, the thermal diffusivity of reference composites of KCl-NaCl-Al was measured as a function of aluminium metal content at room temperature. The thermal conductivity of the reference composites was found to increase with the metallic Al content. The lumped parameter model approach was carried out to discuss the influence of different geometry arrangements of each phase, viz. air, salts and metallic aluminium on the thermal conductivity. Application of the present results to industrial samples indicates that factors such as the interfacial condition of metallic Al particles have to be considered in order to estimate the amount of entrapped Al in the salt cake.

  15. 12th Annual ALS Users' Association Meeting

    SciTech Connect

    Robinson, Arthur L.

    1999-12-17

    Science took the front seat as 219 Advanced Light Source (ALS) users and staff gathered on Monday and Tuesday, October 18 and 19 for the twelfth annual users' meeting. The bulk of the meeting was dedicated to reports on science at the ALS. Packed into two busy days were 31 invited oral presentations and 80 submitted poster presentations, as well as time to visit 24 vendor booths. The oral sessions were dedicated to environmental science, chemical dynamics, biosciences, magnetic materials, and atomic and molecular science. In addition, there was an ALS highlights session that emphasized new results and a session comprising highlights from the young scientists who will carry the ALS into the future.

  16. 17th Annual ALS Users' Association Meeting

    SciTech Connect

    Robinson, Art; Tamura, Lori

    2004-11-29

    It's not exactly Russian roulette, but scheduling October events outdoors is not risk-free, even in usually sunny California. An overflow crowd of more than 400 registered users, ALS staff, and vendors enjoyed a full indoor program featuring science highlights and workshops spread over two and a half days from October 18 to October 20. However, a major storm, heralding the onset of the San Francisco Bay Area rainy season, posed a few weather challenges for the events on the ALS patio.

  17. Continuous germanene layer on Al(111).

    PubMed

    Derivaz, Mickael; Dentel, Didier; Stephan, Régis; Hanf, Marie-Christine; Mehdaoui, Ahmed; Sonnet, Philippe; Pirri, Carmelo

    2015-04-01

    Germanene, a 2D honeycomb structure similar to silicene, has been fabricated on Al(111). The 2D germanene layer covers uniformly the substrate with a large coherence over the Al(111) surface atomic plane. It is characterized by a (3 × 3) superstructure with respect to the substrate lattice, shown by low energy electron diffraction and scanning tunnelling microscopy. First-principles calculations indicate that the Ge atoms accommodate in a very regular atomic configuration with a buckled conformation. PMID:25802988

  18. Metastability in the MgAl2O4-Al2O3 System

    DOE PAGES

    Wilkerson, Kelley R.; Smith, Jeffrey D.; Hemrick, James G.

    2014-07-22

    Aluminum oxide must take a spinel form ( γ-Al2O3) at elevated temperatures in order for extensive solid solution to form between MgAl2O4 and α-Al2O3. The solvus line between MgAl2O4 and Al2O3 has been defined at 79.6 wt% Al2O3 at 1500°C, 83.0 wt% Al2O3 at 1600°C, and 86.5 wt% Al2O3 at 1700°C. A metastable region has been defined at temperatures up to 1700°C which could have significant implications for material processing and properties. Additionally, initial processing could have major implications on final chemistry. The spinel solid solution region has been extended to form an infinite solid solution with Al2O3 at elevatedmore » temperatures. A minimum in melting at 1975°C and a chemistry of 96 wt% Al2O3 rather than a eutectic is present, resulting in no eutectic crystal formation during solidification.« less

  19. Next-generation sequencing of 28 ALS-related genes in a Japanese ALS cohort.

    PubMed

    Nakamura, Ryoichi; Sone, Jun; Atsuta, Naoki; Tohnai, Genki; Watanabe, Hazuki; Yokoi, Daichi; Nakatochi, Masahiro; Watanabe, Hirohisa; Ito, Mizuki; Senda, Jo; Katsuno, Masahisa; Tanaka, Fumiaki; Li, Yuanzhe; Izumi, Yuishin; Morita, Mitsuya; Taniguchi, Akira; Kano, Osamu; Oda, Masaya; Kuwabara, Satoshi; Abe, Koji; Aiba, Ikuko; Okamoto, Koichi; Mizoguchi, Kouichi; Hasegawa, Kazuko; Aoki, Masashi; Hattori, Nobutaka; Tsuji, Shoji; Nakashima, Kenji; Kaji, Ryuji; Sobue, Gen

    2016-03-01

    We investigated the frequency and contribution of variants of the 28 known amyotrophic lateral sclerosis (ALS)-related genes in Japanese ALS patients. We designed a multiplex, polymerase chain reaction-based primer panel to amplify the coding regions of the 28 ALS-related genes and sequenced DNA samples from 257 Japanese ALS patients using an Ion Torrent PGM sequencer. We also performed exome sequencing and identified variants of the 28 genes in an additional 251 ALS patients using an Illumina HiSeq 2000 platform. We identified the known ALS pathogenic variants and predicted the functional properties of novel nonsynonymous variants in silico. These variants were confirmed by Sanger sequencing. Known pathogenic variants were identified in 19 (48.7%) of the 39 familial ALS patients and 14 (3.0%) of the 469 sporadic ALS patients. Thirty-two sporadic ALS patients (6.8%) harbored 1 or 2 novel nonsynonymous variants of ALS-related genes that might be deleterious. This study reports the first extensive genetic screening of Japanese ALS patients. These findings are useful for developing genetic screening and counseling strategies for such patients.

  20. Magnetism in Fe4Al13 and related FeAl intermetallics

    NASA Astrophysics Data System (ADS)

    Chi, Ji; Li, Yang; Gou, Weiping; Goruganti, V.; Rathnayaka, K. D. D.; Ross, Joseph H., Jr.

    2006-03-01

    We report the results of an experimental study of FeAl alloys, including Fe4Al13, FeAl2 and Fe2Al5. By using NMR, dc magnetic susceptibility, and specific heat, we found that Fe4Al13 and Fe2Al5 are non-magnetic with some dilute magnetic moments, while FeAl2 can be characterized as a concentrated local moment system. Fe4Al13 is a decagonal quasicrystal approximant with 102 atoms in its unit cell. The ^27Al NMR spin-lattice relaxation indicates a very narrow pseudogap in the electronic density of states [g(E)] in the vicinity of the Fermi energy. The observations could be fit assuming a parabolic variation of g(E), consistent with observations in other quasicrystals and approximants. NMR lineshape measurements also agree with this analysis, and show that the system is dilute-magnetic, in strong contrast to the FeAl2 ordered intermetallic. We use specific heat to analyze the dilute moment density. This work was supported by the Robert A. Welch Foundation, Grant No. A-1526, by the National Science Foundation (DMR-0103455), and by Texas A&M University through the Telecommunications and Informatics Task Force.

  1. Thermodynamics and surface properties of liquid Al-Ga and Al-Ge alloys

    NASA Astrophysics Data System (ADS)

    Anusionwu, B. C.; Adebayo, G. A.; Madu, C. A.

    2009-11-01

    The surface properties of Al-Ga and Al-Ge liquid alloys have been theoretically investigated at a temperature of 1100 K and 1220 K respectively. For the Al-Ga system, the quasi chemical model for regular alloy and a model for phase segregating alloy systems were applied, while for the Al-Ge system the quasi chemical model for regular and compound forming binary alloys were applied. In the case of Al-Ga, the models for the regular alloys and that for the phase segregating alloys produced the same value of order energy and same values of thermodynamic and surface properties, while for the Al-Ge system, the model for the regular alloy reproduced better the thermodynamic properties of the alloy. The model for the compound forming systems showed a qualitative trend with the measured values of the thermodynamic properties of the Al-Ge alloy and suggests the presence of a weak complex of the form Al2Ge3. The surface concentrations for the alloys show that Ga manifests some level of surface segregation in Al-Ga liquid alloy while the surface concentration of Ge in Al-Ge liquid alloy showed a near Roultian behavior below 0.8 atomic fraction of Ge.

  2. Clinical trials for neuroprotection in ALS.

    PubMed

    Siciliano, G; Carlesi, C; Pasquali, L; Piazza, S; Pietracupa, S; Fornai, F; Ruggieri, S; Murri, L

    2010-07-01

    Owing to uncertainty on the pathogenic mechanisms underlying motor neuron degeneration in amyotrophic lateral sclerosis (ALS) riluzole remains the only available therapy, with only marginal effects on disease survival. Here we review some of the recent advances in the search for disease-modifying drugs for ALS based on their putative neuroprotective effetcs. A number of more or less established agents have recently been investigated also in ALS for their potential role in neuroprotection and relying on antiglutamatergic, antioxidant or antiapoptotic strategies. Among them Talampanel, beta-lactam antibiotics, Coenzyme Q10, and minocycline have been investigated. Progress has also been made in exploiting growth factors for the treatment of ALS, partly due to advances in developing effective delivery systems to the central nervous system. A number of new therapies have also been identified, including a novel class of compounds, such as heat-shock protein co-inducers, which upregulate cell stress responses, and agents promoting autophagy and mitochondriogenesis, such as lithium and rapamycin. More recently, alterations of mRNA processing were described as a pathogenic mechanism in genetically defined forms of ALS, as those related to TDP-43 and FUS-TLS gene mutations. This knowledge is expected to improve our understanding of the pathogenetic mechanism in ALS and developing more effective therapies. PMID:20406180

  3. Modeling of precipitation in Al alloys

    SciTech Connect

    Asta, M.; Foiles, S.M.; Wolfer, W.G.

    1996-10-01

    Objective was the development of a computational model of precipitation from a supersaturated alloy solid solution. The model is based on the formalism of chemical-reaction-rate theory combined with classical descriptions of precipitate thermodynamic properties and a mean-field treatment of diffusion-limited growht and coarsening. For the case of precipitation of Al{sub 3}Sc in supersaturated Al-Sc alloys, it is demonstrated how the model can be used to calculate number densities and size distributions of precipitates as a function of aging time and temperature, including effects of continuous cooling and thermally generated point defects. Application of the model to a specific alloy system requires knowledge of diffusion data, point defect energetics, and thermodynamic properties for bulk phases and interphase interfaces. For interfaces and point defects, thermodynamic data can be difficult to measure and reliable values of defect free energies are often unavailable. For this reason, part of the efforts were devoted to applying semiempirical and first-principles atomistic techniques to the calculation of interfacial and point-defect thermodynamic properties. This report discusses applications for interphase interfaces in the Al-Ag, Al-Sc, and Al-Li alloy systems. We also describe atomistic work aimed at understanding the energetics of vacancy clusters in Al. These clusters serve as sinks for isolated vacancies during aging and their growth can lead to more complex defects, such as dislocation loops, that act as heterogeneous nucleation sites.

  4. Kinetic Monte Carlo of transport processes in Al/AlOx/Au-layers: Impact of defects

    NASA Astrophysics Data System (ADS)

    Weiler, Benedikt; Haeberle, Tobias; Gagliardi, Alessio; Lugli, Paolo

    2016-09-01

    Ultrathin films of alumina were investigated by a compact kMC-model. Experimental jV-curves from Al/AlOx/Au-junctions with plasma- and thermal-grown AlOx were fitted by simulated ones. We found dominant defects at 2.3-2.5 eV below CBM for AlOx with an effective mass mox ∗= 0.35 m0 and a barrier EB ,A l /A l O x≈2.8 eV in agreement with literature. The parameterization is extended to varying defect levels, defect densities, injection barriers, effective masses and the thickness of AlOx. Thus, dominant charge transport processes and implications on the relevance of defects are derived and AlOx parameters are specified which are detrimental for the operation of devices.

  5. Theoretical studies of Ni/sub 3/Al and NiAl with impurities

    SciTech Connect

    Chen, S.P.; Voter, A.F.; Boring, A.M.; Albers, R.C.; Hay, P.J.

    1988-01-01

    Intermetallic compound has been extensively studied because of their superior properties in strength, low creep rate, and high melting point. But most of the systems have room temperature ductility problems, like Ll/sub 2/ and B2 compounds. Both Ll/sub 2/ Ni/sub 3/Al and B2 NiAl exhibit intergranular fracture mode. Understanding grain boundaries in these materials is of particular importance since intergranular fracture limits the applicability of these otherwise promising material. In an effort trying to understand the fracture mechanism, we have used embedded atom potentials to study the properties of Ni/sub 3/Al and NiAl. We also consider the effect of boron, sulfur, and nickel segregation on the strength of grain boundaries in Ni/sub 3/Al and NiAl. 22 refs., 2 figs.

  6. Scattering induced by Al segregation in AlGaN/GaN heterostructures

    SciTech Connect

    Liu, Xiwen; Lu, Yanwu; Ji, Dong

    2015-08-17

    The effect of Al segregation near dislocations on the mobility of two-dimensional electron gas in AlGaN/GaN heterostructure-based high-electron-mobility transistors was investigated. Exponentially varied composition fluctuation was effective in describing Al segregation near dislocations when calculating scattering behavior. Mobility, which was limited by Al segregation surrounding dislocation lines, was calculated to be in the order of 10{sup 3} cm{sup 2}/Vs to 10{sup 6} cm{sup 2}/Vs. Results indicated that the mobility in AlGaN/GaN heterojunction was enhanced upon the reduction of dislocation density at low temperature. This study contributes to generating higher electron mobility in AlGaN/GaN heterojunctions.

  7. Microscopic Properties of Long-Period Ordering in Al-Rich TiAl Alloys

    NASA Astrophysics Data System (ADS)

    Hata, S.; Nakano, T.; Kuwano, N.; Itakura, M.; Matsumura, S.; Umakoshi, Y.

    2008-07-01

    The ordering mechanism of long-period superstructures (LPSs) in Al-rich TiAl alloys has been investigated by high-resolution transmission electron microscopy (HRTEM). The LPSs are classified in terms of arrangements of base clusters with different shapes and compositions formed in Ti-rich (002) layers of L10-TiAl matrix: square Ti4Al, fat rhombus Ti3Al, and lean rhombus Ti2Al type clusters. The HRTEM observations revealed that antiphase boundaries of long-range-ordered LPS domains and short-range-ordered microdomains are constructed by various space-filling arrangements of the base clusters. Such a microscopic property characterized by the base clusters and their arrangements is markedly analogous to that of the {left< {{text{1,1/2,0}}} rightrangle } * special-point ordering alloys such as Ni-Mo.

  8. Epidemiology and neurological complications of infection by the Zika virus: a new emerging neurotropic virus.

    PubMed

    Carod-Artal, Francisco J

    2016-04-01

    Introduccion. El actual brote epidemico por virus Zika se inicio en 2015 y en la actualidad afecta a 31 paises y territorios en America. Se revisan los aspectos epidemiologicos y clinicos asociados con la infeccion por virus Zika. Desarrollo. Desde 2007, 55 paises de America, Asia, Africa y Oceania han detectado transmision local del virus. La actual epidemia ha afectado a casi 1,5 millones de personas en Brasil. El 80% de los casos son asintomaticos. La enfermedad por virus Zika cursa con fiebre, exantema maculopapular, artralgias y conjuntivitis no purulenta. Los sintomas suelen ser autolimitados y duran una semana. Se ha descrito un aumento de la incidencia de los casos de microcefalia, lesiones retinianas y sindrome de Guillain-Barre asociados con el virus Zika. El sindrome de Guillain-Barre asociado al Zika en la Polinesia es una variante axonal motora pura. El ARN del virus Zika se ha identificado en muestras de tejido cerebral, placenta y liquido amniotico de niños con microcefalia y en perdidas fetales de mujeres infectadas por Zika durante el embarazo. Se recomienda realizar la prueba de reaccion en cadena de la polimerasa mediante transcriptasa inversa para detectar ARN virico y pruebas serologicas (IgM ELISA y anticuerpos neutralizantes) para confirmar una infeccion por Zika. El diagnostico diferencial incluye la infeccion por virus dengue y chikungunya. Conclusiones. Existe un conocimiento limitado sobre los mecanismos patogenicos implicados y las consecuencias a largo plazo de la infeccion por virus Zika en adultos y recien nacidos.

  9. Remarks on Peinado et al.'s Analysis of J3Gen

    PubMed Central

    Garcia-Alfaro, Joaquin; Herrera-Joancomartí, Jordi; Melià-Seguí, Joan

    2015-01-01

    Peinado et al. analyzed the security of the J3Gen pseudorandom number generator proposed by Melià-Seguí et al., and claimed weaknesses regarding its security properties. They also presented a deterministic attack based on the decimation of the J3Gen output sequences. We show that the assumptions made by Peinado et al. are not correct and that the proposed deterministic attack against J3Gen does not hold in practice. PMID:25781510

  10. Two-Phase (TiAl+TiCrAl) Coating Alloys for Titanium Aluminides

    NASA Technical Reports Server (NTRS)

    Brady, Michael P. (Inventor); Smialek, James L. (Inventor); Brindley, William J. (Inventor)

    1998-01-01

    A coating for protecting titanium aluminide alloys, including the TiAl gamma + Ti3Al (alpha(sub 2)) class, from oxidative attack and interstitial embrittlement at temperatures up to at least 1000 C. is disclosed. This protective coating consists essentially of titanium, aluminum. and chromium in the following approximate atomic ratio: Ti(41.5-34.5)Al(49-53)Cr(9.5-12.5)

  11. Temperature dependence diode parameters studies of Al/CuPc/n-Si/Al structure

    NASA Astrophysics Data System (ADS)

    Kumar, Ratnesh; Kaur, Ramneek; Sharma, Mamta; Kaur, Maninder; Tripathi, S. K.

    2015-08-01

    This paper presents the fabrication of Al/CuPc/n-Si/Al metal-organic-semiconductor diode. The copper phthalocyanine as organic layer is deposited on Si substrate by thermal evaporation technique. The temperature dependent current-voltage measurements are performed on Al/CuPc/n-Si structure. The important diode parameters i.e. the barrier height and ideality factor have been calculated. The temperature dependence of barrier height and ideality factor has been studied.

  12. The lowest ionization potentials of Al2

    NASA Technical Reports Server (NTRS)

    Bauschlicher, Charles W., Jr.; Barnes, Leslie A.; Taylor, Peter R.

    1988-01-01

    Potential curves for the lowest two electronic states (X 2 sigma g + and A 2 pi u) of Al2(+) were computed using complete active space SCF/multireference CI wave functions and large Gaussian basis sets. The lowest observable vertical ionization potential (to Al2(+) X 2 sigma g +) of the Al2 X 3 pi u ground state is calculated to occur around 6.1 eV, in excellent agreement with the experimental range of 6.0 to 6.42 eV obtained in recent cluster ionization studies by Cox and co-workers. The second vertical ionization potential (to Al2(+) A 2 pi u) occurs near 6.4 eV, also within the experimental range. The adiabatic IP of 5.90 eV is in good agreement with the value of 5.8 to 6.1 eV deduced by Hanley and co-workers from the difference in thresholds between collision induced dissociation processes of Al3(+). The computed IP values are somewhat larger than those deduced from branching ratios in cluster fragmentation experiments by Jarrold and co-workers. The observation of an ionization threshold below 6.42 eV is shown to be incompatible with an Al2 ground electronic state assignment of 3 sigma g -, but the separation between the two lowest states of Al2 is so small that it is likely that both are populated in the experiments, so that this does not provide unambiguous support for the recent theoretical assignment of the ground state as 3 pi u.

  13. Chemical trend of superconducting transition temperature in hole-doped delafossite of CuAlO2, AgAlO2 and AuAlO2

    NASA Astrophysics Data System (ADS)

    Nakanishi, Akitaka; Katayama-Yoshida, Hiroshi

    2012-12-01

    We have performed the first-principles calculations about the superconducting transition temperature Tc of hole-doped delafossite CuAlO2, AgAlO2 and AuAlO2. Calculated Tc are about 50 K (CuAlO2), 40 K (AgAlO2) and 3 K(AuAlO2) at maximum in the optimum hole-doping concentration. The low Tc of AuAlO2 is attributed to the weak electron-phonon interaction caused by the low covalency and heavy atomic mass.

  14. High temperature deformation of NiAl and CoAl

    NASA Technical Reports Server (NTRS)

    Nix, W. D.

    1982-01-01

    The high temperature mechanical properties of the aluminides are reviewed with respect to their potential as high temperature structural materials. It is shown that NiAl and CoAl are substantially stronger than the pure metals Ni and Co at high temperatures and approach the strength of some superalloys, particularly when those superalloys are tested in "weak" directions. The factors that limit and control the high temperature strengths of NiAl and CoAl are examined to provide a basis for the development of intermetallic alloys of this type.

  15. Small Al clusters. II - Structure and binding in Al(n) (n = 2-6, 13)

    NASA Technical Reports Server (NTRS)

    Pettersson, Lars G. M.; Bauschlicher, Charles W., Jr.; Halicioglu, Timur

    1987-01-01

    The structure and stability of aluminum clusters containing up to six atoms have been studied using correlated wave functions and extended basis sets. The lowest energy structure is planar for Al4 and Al5, but three dimensional for Al6. The icosahedral, hcp, fcc, and two planar structures of Al13 were considered at the SCF level. The lowest energy structure is the icosahedron, but the planar structures are fairly low lying even in this case. A simplified description using two- and three-body interaction potentials is found to agree well with the ab initio structures and binding energies.

  16. Synthesis of core–shell AlOOH hollow nanospheres by reacting Al nanoparticles with water

    NASA Astrophysics Data System (ADS)

    Lozhkomoev, A. S.; Glazkova, E. A.; Bakina, O. V.; Lerner, M. I.; Gotman, I.; Gutmanas, E. Y.; Kazantsev, S. O.; Psakhie, S. G.

    2016-05-01

    A novel route for the synthesis of boehmite nanospheres with a hollow core and the shell composed of highly crumpled AlOOH nanosheets by oxidizing Al nanopowder in pure water under mild processing conditions is described. The stepwise events of Al transformation into boehmite are followed by monitoring the pH in the reaction medium. A mechanism of formation of hollow AlOOH nanospheres with a well-defined shape and crystallinity is proposed which includes the hydration of the Al oxide passivation layer, local corrosion of metallic Al accompanied by hydrogen evolution, the rupture of the protective layer, the dissolution of Al from the particle interior and the deposition of AlOOH nanosheets on the outer surface. In contrast to previously reported methods of boehmite nanoparticle synthesis, the proposed method is simple, and environmentally friendly and allows the generation of hydrogen gas as a by-product. Due to their high surface area and high, slit-shaped nanoporosity, the synthesized AlOOH nanostructures hold promise for the development of more effective catalysts, adsorbents, vaccines and drug carriers.

  17. Elastic and piezoelectric properties of AlN and LiAlO2 single crystals.

    PubMed

    Sotnikov, Andrey; Schmidt, Hagen; Weihnacht, Manfred; Smirnova, Elena; Chemekova, Tatiana; Makarov, Yuri

    2010-04-01

    We have successfully grown high-quality AlN piezoelectric single crystal using the sublimation technique. Transparent crack-free boules of approximately 15 mm in diameter and 25 mm in length along the [0001] direction were obtained, with coloring from amber to dark brown depending on growth temperature. Full sets of material parameters of grown AlN and commercially available LiAlO(2) bulk crystals were measured at room temperature. Temperature coefficients of the material parameters of LiAlO(2) were also obtained in a temperature range from -70 to +50 degrees C.

  18. Synthesis of core-shell AlOOH hollow nanospheres by reacting Al nanoparticles with water

    NASA Astrophysics Data System (ADS)

    Lozhkomoev, A. S.; Glazkova, E. A.; Bakina, O. V.; Lerner, M. I.; Gotman, I.; Gutmanas, E. Y.; Kazantsev, S. O.; Psakhie, S. G.

    2016-05-01

    A novel route for the synthesis of boehmite nanospheres with a hollow core and the shell composed of highly crumpled AlOOH nanosheets by oxidizing Al nanopowder in pure water under mild processing conditions is described. The stepwise events of Al transformation into boehmite are followed by monitoring the pH in the reaction medium. A mechanism of formation of hollow AlOOH nanospheres with a well-defined shape and crystallinity is proposed which includes the hydration of the Al oxide passivation layer, local corrosion of metallic Al accompanied by hydrogen evolution, the rupture of the protective layer, the dissolution of Al from the particle interior and the deposition of AlOOH nanosheets on the outer surface. In contrast to previously reported methods of boehmite nanoparticle synthesis, the proposed method is simple, and environmentally friendly and allows the generation of hydrogen gas as a by-product. Due to their high surface area and high, slit-shaped nanoporosity, the synthesized AlOOH nanostructures hold promise for the development of more effective catalysts, adsorbents, vaccines and drug carriers.

  19. Synthesis of core-shell AlOOH hollow nanospheres by reacting Al nanoparticles with water.

    PubMed

    Lozhkomoev, A S; Glazkova, E A; Bakina, O V; Lerner, M I; Gotman, I; Gutmanas, E Y; Kazantsev, S O; Psakhie, S G

    2016-05-20

    A novel route for the synthesis of boehmite nanospheres with a hollow core and the shell composed of highly crumpled AlOOH nanosheets by oxidizing Al nanopowder in pure water under mild processing conditions is described. The stepwise events of Al transformation into boehmite are followed by monitoring the pH in the reaction medium. A mechanism of formation of hollow AlOOH nanospheres with a well-defined shape and crystallinity is proposed which includes the hydration of the Al oxide passivation layer, local corrosion of metallic Al accompanied by hydrogen evolution, the rupture of the protective layer, the dissolution of Al from the particle interior and the deposition of AlOOH nanosheets on the outer surface. In contrast to previously reported methods of boehmite nanoparticle synthesis, the proposed method is simple, and environmentally friendly and allows the generation of hydrogen gas as a by-product. Due to their high surface area and high, slit-shaped nanoporosity, the synthesized AlOOH nanostructures hold promise for the development of more effective catalysts, adsorbents, vaccines and drug carriers. PMID:27053603

  20. Al-matrix composite materials reinforced by Al-Cu-Fe particles

    NASA Astrophysics Data System (ADS)

    Bonneville, J.; Laplanche, G.; Joulain, A.; Gauthier-Brunet, V.; Dubois, S.

    2010-07-01

    Al-matrix material composites were produced using hot isostatic pressing technique, starting with pure Al and icosahedral (i) Al-Cu-Fe powders. Depending on the processing temperature, the final reinforcement particles are either still of the initial i-phase or transformed into the tetragonal ω-Al00.70Cu0.20Fe0.10 crystalline phase. Compression tests performed in the temperature range 293K - 823K on the two types of composite, i.e. Al/i and Al/ω, indicate that the flow stress of both composites is strongly temperature dependent and exhibit distinct regimes with increasing temperature. Differences exist between the two composites, in particul ar in yield stress values. In the low temperatureregime (T <= 570K), the yield stress of the Al/ω composite is nearly 75% higher than that of the Al/i composite, while for T > 570K both composites exhibit similar yield stress values. The results are interpreted in terms of load transfer contribution between the matrix and the reinforcement particles and elementary dislocation mechanisms in the Al matrix.

  1. The new structure type Gd3Ni7Al14.

    PubMed

    Pukas, Svitlana; Gladyshevskii, Roman

    2015-11-01

    The crystal structure of Gd3Ni7Al14 (trigadolinium heptanickel tetradecaaluminide) belongs to a family of two-layer structures and can be described as an assembly of interpenetrating centred straight prisms. For the Ni atoms, trigonal prisms (Al4Gd2 and Al6) are observed, the Al atoms are inside tetragonal (Ni2Al2Gd4, Ni2Al4Gd2, Al4Gd4, Ni4Al4 and Al8) and pentagonal (Ni4Al6 and Al10) prisms, while the Gd atoms are at the centres of pentagonal (Ni4Al6) and hexagonal (Ni4Al8) prisms. In each case, the true coordination polyhedron is a capped prism, also including atoms from the same layer. The structural features of Gd3Ni7Al14 are similar to those of the intermetallides PrNi2Al3 and ZrNiAl. In all these structures, Ni-centred trigonal prisms form infinite columns via common triangular faces. The columns share prism edges and form a three-dimensional framework with six-membered rings in the (001) plane in the case of the PrNi2Al3 and ZrNiAl types. In the case of Gd3Ni7Al14, six-membered rings are also observed, but only two-thirds of the rings are interconnected via prism edges. PMID:26524174

  2. The new structure type Gd3Ni7Al14.

    PubMed

    Pukas, Svitlana; Gladyshevskii, Roman

    2015-11-01

    The crystal structure of Gd3Ni7Al14 (trigadolinium heptanickel tetradecaaluminide) belongs to a family of two-layer structures and can be described as an assembly of interpenetrating centred straight prisms. For the Ni atoms, trigonal prisms (Al4Gd2 and Al6) are observed, the Al atoms are inside tetragonal (Ni2Al2Gd4, Ni2Al4Gd2, Al4Gd4, Ni4Al4 and Al8) and pentagonal (Ni4Al6 and Al10) prisms, while the Gd atoms are at the centres of pentagonal (Ni4Al6) and hexagonal (Ni4Al8) prisms. In each case, the true coordination polyhedron is a capped prism, also including atoms from the same layer. The structural features of Gd3Ni7Al14 are similar to those of the intermetallides PrNi2Al3 and ZrNiAl. In all these structures, Ni-centred trigonal prisms form infinite columns via common triangular faces. The columns share prism edges and form a three-dimensional framework with six-membered rings in the (001) plane in the case of the PrNi2Al3 and ZrNiAl types. In the case of Gd3Ni7Al14, six-membered rings are also observed, but only two-thirds of the rings are interconnected via prism edges.

  3. Evidence that 26Al Did Not Melt Asteroids

    NASA Astrophysics Data System (ADS)

    Wasson, J. T.

    2016-08-01

    26Al/27Al initial ratios in achondrites are much lower than expected if 26Al was the only heat source responsible for melting the parental materials. Impacts provided a substantial fraction of the heat.

  4. Exothermic reactions in Co/Al nanolaminates

    NASA Astrophysics Data System (ADS)

    Adams, D. P.; Hodges, V. C.; Bai, M. M.; Jones, E.; Rodriguez, M. A.; Buchheit, T.; Moore, J. J.

    2008-08-01

    Vapor-deposited Co/Al nanolaminates having a net equiatomic stoichiometry exhibited rapid, high-temperature synthesis. When ignited locally, 7.5-μm-thick Co/Al foils were characterized by self-sustained propagating reactions with flame front speeds between ˜0.5 and 9 m/s. Speed was largely affected by bilayer thickness and premixed volume fraction, consistent with previous studies of other exothermic multilayers. Cobalt aluminide foils reacted by self-propagating synthesis developed a variety of rough surface morphologies characterized by a large amplitude (peak-to-valley ˜1.0 μm), multiperiod, wavelike structure. High-temperature reaction was also stimulated by rapid global heating. Tests revealed low ignition temperatures (Tig)˜240-460 °C compared with previous Co-Al powder reactant experiments. Ignition temperature was influenced by bilayer thickness. All foils ignited by global heating and those reacted in a self-propagating mode developed a single-phase B2 (CsCl-type) crystal structure. Thick Co/Al nanolaminates were used successfully to join a few different materials including metalized Al2O3 (by soldering) and polyetheretherketone (by direct attachment).

  5. Oscillatory surface relaxations in Ni, Al, and their ordered alloys

    SciTech Connect

    Chen, S.P.; Voter, A.F.; Srolovitz, D.J.

    1986-09-15

    Results from simulations of Ni, Al, Ni/sub 3/Al, and NiAl show long-range, oscillatory surface relaxations that decay exponentially into the bulk. Pure fcc Ni and Al have oscillation periods that are close to the nearest-neighbor distance, independent of crystal face. This is shown to be due to surface smoothing and steric effects. In Ni/sub 3/Al and NiAl, the surface planes are rippled, with the Ni-Ni and Al-Al interlayer spacings oscillating 180/sup 0/ out of phase. Very good agreement between our results and experimentally measured atomic relaxations is obtained.

  6. The mechanical properties of FeAl

    SciTech Connect

    Baker, I.; George, E.P.

    1996-12-31

    Only in the last few years has progress been made in obtaining reproducible mechanical properties data for FeAl. Two sets of observations are the foundation of this progress. The first is that the large vacancy concentrations that exist in FeAl at high temperature are easily retained at low temperature and that these strongly affect the low-temperature mechanical properties. The second is that RT ductility is adversely affected by water vapor. Purpose of this paper is not to present a comprehensive overview of the mechanical properties of FeAl but rather to highlight our understanding of key phenomena and to show how an understanding of the factors which control the yield strength and fracture behavior has followed the discovery of the above two effects. 87 refs, 9 figs.

  7. Boron strengthening in FeAl

    SciTech Connect

    Baker, I.; Li, X.; Xiao, H.; Klein, O.; Nelson, C.; Carleton, R.L.; George, E.P.

    1998-11-01

    The effect of boron on the strength of B2-structured FeAl is considered as a function of composition, grain size and temperature. Boron does not affect the concentrations of antisite atoms or vacancies present, with the former increasing and the latter decreasing with increasing deviation from the stoichiometric composition. When vacancies are absent, the strength increase per at. % B per unit lattice strain, {Delta}{sigma}/({Delta}c x {epsilon}) increases with increasing aluminum concentration, but when vacancies are present (>45 at. % Al), {Delta}{sigma}/({Delta}c x {epsilon}) decreases again. Boron increases grain size strengthening in FeAl. B strengthening is roughly independent of temperature up to the yield strength peak but above the point, when diffusion-assisted deformation occurs, boron strengthening increases dramatically.

  8. [Al-Biruni--a universal scientist].

    PubMed

    Kujundzić, E; Masić, I

    1999-01-01

    Al-Biruni's was of Persian descent. He was born in Horesmiya and had studied mathematics, history and medicine. Acquiring knowledge from these sciences, he wrote an outstanding work on chronology of several nations and devoted it to Ziyarit ruler Kabus. He made a chronological overview of calendars from many nations, including Persians, Greeks, Egyptians, Jews, Melkitian and Nestorian Christians, Sabeyaans as well as the old Arabs. Data presented in the work, according to the later authors, were taken from very reliable sources. He was contemporary of Ibn-Sina, and thanks to their friendship, they have discussed very much miscellaneous topics. He belonged to the group of scholars, taken by Gaznevian Soultan Mahmud to a long journey to India. Afterwards Al-Biruni wrote and published detailed work "Description of India"--a work on cultural history of India. Due to excellent abilities of Al-Biruni as a philosopher and scholar, there are still significant and reliable notes about buddhistic philosophy, structure of castes and Brahmans' life style. In this Al-Biruni's masterpiece, there are many comparative analysis of Suffism and certain Indian philosophical methods. Al-Biruni's most important work is "Pharmacopoeia"--"Kitab al-saydala", which brilliantly describes all medicaments. This work has been published in many languages. He also wrote few works on astronomy and astrology. In those works he has explained some astrological events through scientific approach in a such peculiar way that nobody has ever explained before. He was also interested in sciences like geology, mineralology, geography, mathematics, psychology and many others. PMID:10386051

  9. Individual two level fluctuators in the tunneling conductance of Al/AlOx/Al Josephson junctions for superconducting qubits

    NASA Astrophysics Data System (ADS)

    Nugroho, Christopher; Orlyanchik, Vladimir; van Harlingen, Dale

    2014-03-01

    Two level system (TLS) defects in AlOx tunnel barriers can lead to low-frequency 1 / f critical current noise and losses in coherent superconducting circuits. Understanding the nature of these defects and how to eliminate them are critical in order to achieve ultra-long coherence times. We present measurements of the tunneling conductance of ultrasmall, A <(100nm) 2 , Al/AlOx/Al shadow evaporated junctions. The tunneling conductance of these junctions exhibits several isolated TLSs, which permitted the detailed analysis of the individual switching rates and behavior of the TLSs. We have studied the thermal activation behavior of these TLSs, and in some cases observe a crossover into quantum-limited tunneling at lower temperatures. Tracking the TLS switching rates as a function of the applied voltage bias provides an estimate of the TLS charge dipole moment. In some quantum tunneling limited TLSs we have observed a non-equilibrium enhancement of the switching rates that cannot be explained by simple dissipative heating of the TLSs. Further investigations into these TLS defects may lead to the identification of their physical origins and strategies to eliminate them. Research funded by the Intelligence Advanced Research Projects Activity (IARPA).

  10. Complementary and Alternative Therapies in ALS

    PubMed Central

    Bedlack, Richard S.; Joyce, Nanette; Carter, Gregory T.; Pagononi, Sabrina; Karam, Chafic

    2015-01-01

    Synopsis Given the severity of their illness and lack of effective disease modifying agents, it is not surprising that most patients with ALS consider trying complementary and alternative therapies. Some of the most commonly considered alternative therapies include special diets, nutritional supplements, cannabis, acupuncture, chelation and energy healing. This chapter reviews these in detail. We also describe 3 models by which physicians may frame discussions about alternative therapies: paternalism, autonomy and shared decision making. Finally, we review a program called ALSUntangled which using shared shared decision making to review alternative therapies for ALS. PMID:26515629

  11. A Comment on Eerland et al. (2016).

    PubMed

    Hart, William

    2016-03-01

    People imagine criminal behavior in greater detail and ascribe more blame to criminals when a description of a crime is written using the imperfective verb aspect (Hart & Albarracín, 2011, Experiment 3). An initiative to reproduce these findings conducted by Eerland et al. (2016) yielded a lack of replication. Eerland et al. have suggested that the original effect was likely driven by a Type I error produced by methodological aspects of the experiment, including low sample size. Recent conceptual replications of Hart and Albarracín's effect (Sherrill, Eerland, Zwaan, & Magliano, 2015), however, might imply the effect could be contingent on sample characteristics. PMID:26993281

  12. Fabrication and Characterization of AL-AL4C3 Nanocomposite by Mechanical Alloying

    NASA Astrophysics Data System (ADS)

    Mohammad Sharifi, E.; Enayati, M. H.; Karimzadeh, F.

    Aluminum carbide (Al4C3) seems to be an ideal reinforcement for producing aluminum matrix composites. Al4C3 has high hardness and shear strength as well as a high melting point. The dispersion of Al4C3 particles in the matrix produces a pinning effect that reduces aluminum grain growth, which improves the mechanical properties. In this study, aluminum powders were mixed with 4.5 wt.% graphite and mechanically alloyed using a high-energy ball mill in order to produce Al-Al4C3 nanocomposite. The structural evaluation of powder particles after different milling times was studied by X-ray diffractometry, scanning electron microscopy and microhardness measurements. The aluminum crystallite size estimated with broadening of XRD peaks by Williamson-Hall formula. XRD results suggested that the grain size of aluminum decreased to nanometer range (30 nm) during ball milling. No Al4C3 formed during the mechanical alloying process. Milled powders were then annealed at 300-600 °C for 1 h under argon atmosphere. Annealing at temperatures higher than 300 °C led to formation of Al4C3 phase which increased as annealing temperature increased. Aluminum grain size remained in nanometer range after annealing process. The microhardness of powder particles increased after annealing. Both effects are due to the formation of nanosized Al4C3 particles.

  13. Melting, Processing, and Properties of Disordered Fe-Al and Fe-Al-C Based Alloys

    NASA Astrophysics Data System (ADS)

    Satya Prasad, V. V.; Khaple, Shivkumar; Baligidad, R. G.

    2014-09-01

    This article presents a part of the research work conducted in our laboratory to develop lightweight steels based on Fe-Al alloys containing 7 wt.% and 9 wt.% aluminum for construction of advanced lightweight ground transportation systems, such as automotive vehicles and heavy-haul truck, and for civil engineering construction, such as bridges, tunnels, and buildings. The melting and casting of sound, porosity-free ingots of Fe-Al-based alloys was accomplished by a newly developed cost-effective technique. The technique consists of using a special flux cover and proprietary charging schedule during air induction melting. These alloys were also produced using a vacuum induction melting (VIM) process for comparison purposes. The effect of aluminum (7 wt.% and 9 wt.%) on melting, processing, and properties of disordered solid solution Fe-Al alloys has been studied in detail. Fe-7 wt.% Al alloy could be produced using air induction melting with a flux cover with the properties comparable to the alloy produced through the VIM route. This material could be further processed through hot and cold working to produce sheets and thin foils. The cold-rolled and annealed sheet exhibited excellent room-temperature ductility. The role of carbon in Fe-7 wt.% Al alloys has also been examined. The results indicate that Fe-Al and Fe-Al-C alloys containing about 7 wt.% Al are potential lightweight steels.

  14. Transport mechanisms of leakage current in Al2O3/InAlAs MOS capacitors

    NASA Astrophysics Data System (ADS)

    Jin, Chengji; Lu, Hongliang; Zhang, Yimen; Zhang, Yuming; Guan, He; Wu, Lifan; Lu, Bin; Liu, Chen

    2016-09-01

    An Al2O3 layer is inserted between the InAlAs layer and the metal gate in InAs/AlSb HEMTs to suppress the leakage current. The transport mechanisms of leakage current in Al2O3/InAlAs metal-oxide-semiconductor (MOS) capacitors at both positive and negative biases at different temperatures ranging from 10 °C to 70 °C are investigated. For positive bias, the leakage current is dominated by Schottky emission. Based on the fitted straight lines, the relative dielectric constant of Al2O3 and the barrier height between Al2O3 and InAlAs are extracted. However, for negative bias, the leakage current is dominated by Frenkel-Poole (F-P) emission and the depth of the trap energy level from the conduction band (ϕt) is extracted. Furthermore, authors explain the reason why the dominating mechanisms at positive and negative biases are different.

  15. Electronic circuits having NiAl and Ni.sub.3 Al substrates

    DOEpatents

    Deevi, Seetharama C.; Sikka, Vinod K.

    1999-01-01

    An electronic circuit component having improved mechanical properties and thermal conductivity comprises NiAl and/or Ni.sub.3 Al, upon which an alumina layer is formed prior to applying the conductive elements. Additional layers of copper-aluminum alloy or copper further improve mechanical strength and thermal conductivity.

  16. Directional Solidification and Mechanical Properties of NiAl-NiAlTa Alloys

    NASA Technical Reports Server (NTRS)

    Johnson, D. R.; Chen, X. F.; Oliver, B. F.; Noebe, R. D.; Whittenberger, J. D.

    1995-01-01

    Directional solidification of eutectic alloys is a promising technique for producing in-situ composite materials exhibiting a balance of properties. Consequently, the microstructure, creep strength and fracture toughness of directionally solidified NiAl-NiAlTa alloys were investigated. Directional solidification was performed by containerless processing techniques to minimize alloy contamination. The eutectic composition was found to be NiAl-15.5 at% Ta and well-aligned microstructures were produced at this composition. A near-eutectic alloy of NiAl-14.5Ta was also investigated. Directional solidification of the near-eutectic composition resulted in microstructures consisting of NiAl dendrites surrounded by aligned eutectic regions. The off-eutectic alloy exhibited promising compressive creep strengths compared to other NiAl-based intermetallics, while preliminary testing indicated that the eutectic alloy was competitive with Ni-base single crystal superalloys. The room temperature toughness of these two-phase alloys was similar to that of polycrystalline NiAl even with the presence of the brittle Laves phase NiAlTa.

  17. Characterization of nanophase Al-oxide/Al powders by electron energy-loss spectroscopy.

    PubMed

    Fernández; Sánchez-López; Caballero; Martin; Vacher; Ponsonnet

    1998-08-01

    Al nanoparticles were prepared by the inert gas condensation method. After passivation with oxygen and air exposure we obtained a powdered sample of an Al-oxide/Al nanocomposite material. In the present paper we describe the use of the electron energy-loss spectroscopy (EELS) technique in a transmission electron microscope to characterize such nanostructured powders compared with a microcrystalline commercial aluminium foil. Energy-filtered images showed the presence of an alumina overlayer of approximately 4 nm covering the aluminium nanoparticles (23 nm in diameter). EELS analysis enabled us to determine the total amount of Al2O3 and metallic Al and the structure of the alumina passivation overlayer in the sample. In particular, the extended energy-loss fine structure analysis of the data showed a major presence of Al tetrahedrally coordinated with oxygen in the alumina passivation layer of Al nanoparticles instead of the octahedral coordination found for a conventional Al foil. This surprising effect has been attributed to the nanoscopic character of the grains. The analysis of the electron-loss near-edge structure also determines the presence of a certain degree of aggregation in this kind of powdered sample as result of the coalescence of the nanocrystalline grains. The procedure presented here may have the potential to solve other problems during characterization of nanostructured materials.

  18. AlN homoepitaxial growth on sublimation-AlN substrate by low-pressure HVPE

    NASA Astrophysics Data System (ADS)

    Nomura, Takuya; Okumura, Kenta; Miyake, Hideto; Hiramatsu, Kazumasa; Eryu, Osamu; Yamada, Yoichi

    2012-07-01

    Crack-free thick AlN layers with low impurity concentrations were grown on free-standing AlN substrates fabricated by a sublimation method. Cracks due to tensile stresses were generated in the overgrowth layer when using on-axis AlN (0 0 0 1) substrates, as indicated by Raman scattering spectroscopy. In contrast, cracks were not generated when using 5° off-angle AlN (0 0 0 1) substrates. High crystalline quality was indicated by X-ray rocking curve (XRC) analysis. The full width at half maximum (FWHM) values of the (0 0 0 2) and (1 0-1 0) diffractions were 277 and 306 arcsec, respectively. Secondary ion mass spectrometry (SIMS) measurements indicated that the Si and C impurity concentrations were reduced to half of those in the sublimation-grown AlN substrates.

  19. Al/sub 2/O/sub 3/ adherence on CoCrAl alloys

    SciTech Connect

    Kingsley, L.M.

    1980-04-01

    Adherence of protective oxides on NiCrAl and CoCrAl superalloys has been promoted by a dispersion of a highly oxygen reactive element or its oxide being produced within the protection system. Two aspects of this subject are investigated here: the use of Al/sub 2/O/sub 3/ as both the dispersion and protective oxide; and the production of an HfO/sub 2/ dispersion while simultaneously aluminizing the alloy. It was found that an Al/sub 2/O/sub 3/ dispersion will act to promote the adherence of an external scale of Al/sub 2/O/sub 3/ to a degree comparable to previously tested dispersions and an HfO/sub 2/ dispersion comparable to that produced by a Rhines pack treatment is produced during aluminization.

  20. Bonding characters of Al-containing bulk metallic glasses studied by 27Al NMR.

    PubMed

    Xi, X K; Sandor, M T; Wang, H J; Wang, J Q; Wang, W H; Wu, Y

    2011-03-23

    We report very small (27)Al metallic shifts in a series of Cu-Zr-Al bulk metallic glasses. This observation and the Korringa type of spin-lattice relaxation behavior suggest that s-character wavefunctions weakly participate in bonding and opens the possibility of enhanced covalency (pd hybridization)