Sample records for antinuclear antibody positive

  1. Antinuclear Antibodies (ANA)

    MedlinePLUS

    ... itself which can lead to autoimmune diseases, including lupus , scleroderma , Sjögren's syndrome , polymyositis/ dermatomyositis, mixed connective tissue disease, drug-induced lupus, and autoimmune hepatitis. A positive ANA can also ...

  2. Antinuclear antibodies in rosacea patients

    PubMed Central

    Salamon, Ma?gorzata; McCauliffe, Daniel; Sysa-J?drzejowska, Anna

    2013-01-01

    Introduction Rosacea is a common inflammatory disorder, characterized by a spectrum of facial manifestations. The clinical similarity to other dermatoses, like lupus erythematosus, might lead to misdiagnosis, particularly in patients with elevated antinuclear antibody titers. Aim To assess the frequency, titer and specificity of antinuclear antibodies in rosacea patients and correlate these findings with clinical features. Material and methods The study included 101 rosacea patients and 26 sex- and age-matched controls. Immunofluorescence antinuclear antibody testing was performed on HEp-2 substrates. Patients’ sera with ANA titers of 1 : 160 or higher were evaluated by Euroline analysis. Results Over a half (53.5%) of rosacea patients had an ANA titer greater than or equal to 1 : 160. Within this group 13.86% had a titer of 1 : 320, 8.91% had a titer of 1 : 640, and 6.93% had a titer of 1 : 1,280 or higher. The specificity of these antibodies could not be identified. Elevated ANA titers were present more often in women (55.8%) than in men (44.15%). Only two of 26 healthy volunteers had elevated ANA titers. One had a titer of 1 : 160 and the other of 1 : 320. During a two-year observation period, after the initial ANA testing, none of the patients with ANA titers above 1 : 640 developed an apparent autoimmune disorder. Conclusions Elevated ANA titers are commonly found in rosacea patients, what with simultaneously existing facial erythema and photosensitivity might lead to misdiagnosis of lupus erythematosus. Clinicians should beware of these findings to avoid misdiagnosing lupus erythematosus in rosacea patients with elevated ANA titers. PMID:24278039

  3. Antinuclear antibodies in arthritic and nonarthritic children with uveitis.

    PubMed

    Rosenberg, A M; Romanchuk, K G

    1990-01-01

    We determined whether a positive test for antinuclear antibodies correlated with uveitis only in children with juvenile rheumatoid arthritis (JRA) or whether they also represented a serologic marker for isolated idiopathic chronic uveitis in children. We conclude that the immunopathogenesis of uveitis associated with JRA is different from that of idiopathic chronic uveitis. PMID:2313675

  4. High Prevalence of Antinuclear Antibodies in Children with Thyroid Autoimmunity

    PubMed Central

    Segni, Maria; Pucarelli, Ida; Truglia, Simona; Turriziani, Ilaria; Serafinelli, Chiara; Conti, Fabrizio

    2014-01-01

    Background. Antinuclear antibodies (ANA) are a hallmark of many autoimmune diseases and can be detected many years before disease onset. Autoimmune thyroid diseases (AITD) are frequently associated with other organ- and non-organ-specific autoimmune disorders. Objectives. To assess the prevalence of ANA in pediatric patients with AITD and their clinical correlations. Methods. Ninety-three consecutive pediatric patients with AITD were enrolled (86 children with chronic lymphocytic thyroiditis and 7 with Graves' disease). ANA, anti-double DNA (anti-dsDNA) antibodies, anti-extractable nuclear antigen (anti-ENA), anti-cyclic citrullinated peptide antibodies (anti-CCP), and rheumatoid factor (RF) was obtained. Signs and symptoms potentially related to rheumatic diseases in children were investigated by a questionnaire. Results. ANA positivity was found in 66/93 children (71%), anti-ENA in 4/93 (4.3%), anti-dsDNA in 1/93 (1.1%), RF in 3/93 (3.2%), and anti-CCP in none. No significant differences were found between the ANA-positive and ANA-negative groups with respect to age, sex, L-thyroxine treatment, or prevalence of other autoimmune diseases. Overall, parental autoimmunity was found in 23%. Conclusions. ANA positivity was demonstrated in 71% of children with AITD. ANA positivity was not related to overt immune-rheumatic diseases. However, because the positivity of ANA can occur even many years before the onset of systemic autoimmune diseases, prospective studies are warranted. PMID:24741574

  5. Antinuclear antibody and rheumatoid factor in silica-exposed workers.

    PubMed

    Aminian, Omid; Sharifian, Seyed Akbar; Mehrdad, Ramin; Haghighi, Khosro Sadeghniyat; Mazaheri, Maria

    2009-06-01

    A lot of workers in industries such as foundry, stonecutting, and sandblasting are exposed to higher than permissible levels of crystalline silica. Various alterations in humoral immune function have been reported in silicosis patients and workers exposed to silica dust. The aim of this study was to measure antinuclear antibody (ANA) and rheumatoid factor (RF) levels in foundry workers exposed to silica and to compare them with a control group without such exposure. ANA and RF were measured in 78 exposed and 73 non-exposed workers, and standard statistical methods were used to compare them. The two groups did not significantly differ in age and smoking. Mean work duration of the exposed and non-exposed workers was (14.9+/-4.72) years and (12.41+/-6.3) years, respectively. Ten exposed workers had silicosis. ANA was negative in all workers in either group. Its mean titer did not differ significantly between the exposed and control workers [(0.39+/-0.15) IU mL(-1) vs. (0.36+/-0.17) IU mL(-1), respectively]. RF was positive in two workers of each group. Other studies have reported an increase in ANA and RF associated with exposure to silica dust and silicosis.In contrast, our study suggests that exposure to silica dust does not increase the level of ANA and RF in exposed workers. PMID:19581212

  6. Intracellular and circulating neuronal antinuclear antibodies in human epilepsy

    PubMed Central

    Iffland, Philip H.; Carvalho-Tavares, Juliana; Trigunaite, Abhishek; Man, Shumei; Rasmussen, Peter; Alexopoulos, Andreas; Ghosh, Chaitali; Jørgensen, Trine N.; Janigro, Damir

    2013-01-01

    There are overwhelming data supporting the inflammatory origin of some epilepsies (e.g., Rasmussen's encephalitis and limbic encephalitis). Inflammatory epilepsies with an autoimmune component are characterized by autoantibodies against membrane-bound, intracellular or secreted proteins (e.g., voltage gated potassium channels). Comparably, little is known regarding autoantibodies targeting nuclear antigen. We tested the hypothesis that in addition to known epilepsy-related autoantigens, human brain tissue and serum from patients with epilepsy contain autoantibodies recognizing nuclear targets. We also determined the specific nuclear proteins acting as autoantigen in patients with epilepsy. Brain tissue samples were obtained from patients undergoing brain resections to treat refractory seizures, from brain with arteriovenous malformations or from post-mortem multiple sclerosis brain. Patients with epilepsy had no known history of autoimmune disease and were not diagnosed with autoimmune epilepsy. Tissue was processed for immunohistochemical staining. We also obtained subcellular fractions to extract intracellular IgGs. After separating nuclear antibody-antigen complexes, the purified autoantigen was analyzed by mass spectrometry. Western blots using autoantigen or total histones were probed to detect the presence of antinuclear antibodies in the serum of patients with epilepsy. Additionally, HEp-2 assays and antinuclear antibody ELISA were used to detect the staining pattern and specific presence of antinuclear antibodies in serum of patients with epilepsy. Brain regions from patients with epilepsy characterized by blood-brain barrier disruption (visualized by extravasated albumin) contained extravasated IgGs. Intracellular antibodies were found in epilepsy (n=13/13) but not in multiple sclerosis brain (n= 4/4). In brain from patients with epilepsy, neurons displayed higher levels of nuclear IgGs compared to glia. IgG colocalized with extravasated albumin. All subcellular fractions from brain resections of patients with epilepsy contained extravasated IgGs (n=10/10), but epileptogenic cortex, where seizures originated from, displayed the highest levels of chromatin-bound IgGs. In the nuclear IgG pool, anti-histone autoantibodies were identified by two independent immunodetection methods. HEp-2 assay and ELISA confirmed the presence of anti-histone (n= 5/8) and anti-chromatin antibodies in serum from patients with epilepsy We developed a multi-step approach to unmask autoantigens in the brain and sera of patients with epilepsy. This approach revealed antigen-bound antinuclear antibodies in neurons and free antinuclear IgGs in serum of patients with epilepsy. Conditions with blood-brain barrier disruption but not seizures, were characterized by extravasated but not chromatin-bound IgGs. Our results show that the pool of intracellular IgG in brain of patients with epilepsy consists of nucleus-specific autoantibodies targeting chromatin and histones. Seizures may be the trigger of neuronal uptake of antinuclear antibodies. PMID:23880401

  7. Antinuclear antibodies and elevated anti-Epstein-Barr virus titers in cancer patients.

    PubMed Central

    McCormick, N K; McCormick, K J; Trentin, J J

    1976-01-01

    One and one-half percent of human sera from patients seen at a clinic for treatment of cancer contained antibodies to the nuclei of chick kidney cells by indirect immunofluorescence tests. In the group of sera containing antinuclear antibodies, the geometric mean titer to Epstein-Barr virus (EBV) capsid antigen was significantly elevated. Sera obtained from normal adults or from patients with similar histological types of tumors that possessed no antinuclear antibodies contained lower levels of anti-EBV antibodies. The elevated titers to EBV were correlated with the presence or absence of antinuclear antibodies and not with a particular type or site of neoplastic disease. PMID:178605

  8. Prevalence and clinical significance of antinuclear antibodies in Iranian women with unexplained recurrent miscarriage

    PubMed Central

    Molazadeh, Morteza; Karimzadeh, Hadi; Azizi, Mohammad R

    2014-01-01

    Background: Antinuclear antibodies (ANAs) in women with recurrent miscarriage have been reported. The presence of moderate to high titers of these antibodies represents an autoimmune condition that can endanger the health of the fetus in pregnant women. Objective: In this study, we evaluated the prevalence of ANAs in Iranian women with a history of two or more unexplained abortion. Materials and Methods: 560 women with unexplained recurrent miscarriage and 560 healthy controls accounted for this study over a period of 13 months. ANAs were detected by indirect immunofluorescence technique. Results: ANAs were detected in 74 of 560 (13.21%) patient with recurrent miscarriage, and in only 5 of 560 (0.9%) controls (p<0.001). ANA positivity was generally found with low-positive results (1.40-1.80) in about 38% of positive cases, whereas moderate titres (1.160-1.320) and high titres (>1.640) were seen in about 46% and 16% of cases respectively. Finally evaluating of microscopic ANA patterns revealed that about half of positive cases had antibodies against DNA- histone complex, associated with systemic lupus erythematosus disease. Conclusion: Antinuclear antibodies are not uncommon in women with unexplained recurrent miscarriage, suggesting the possible role of an autoimmune disorder on abortion, at least in a subgroup of patients. PMID:24799884

  9. Autoantibodies to lens epithelium-derived growth factor/transcription co-activator P75 (LEDGF/P75) in children with chronic nonspecific complaints and with positive antinuclear antibodies.

    PubMed

    Kuwabara, Natsuko; Itoh, Yasuhiko; Igarshi, Tohru; Fukunaga, Yoshitaka

    2009-09-01

    Autoimmune fatigue syndrome (AIFS) is characterized by chronic nonspecific complaints, consistently positive antinuclear antibodies (ANA), and lack of alternate medical explanations. A newly recognized antibody, named anti-Sa, was detected in approximately 40% of the patients by Western blot (WB) using HeLa extract. Some patients with AIFS later develop chronic fatigue syndrome (CFS), and most of them are positive for anti-Sa. On the other hand, Muro et al. reported anti-DFS70 in patients with CFS. Anti-Sa and anti-DFS70 were turned out to be same specificities by exchanging studies of blind sera. The target antigen of anti-DFS70 was identified as lens epithelium derived growth factor/transcription co-activator p75 (LEDGF/p75). The objectives of this study are to confirm whether the target antigen of anti-Sa is also LEDGF/p75, and to develop ELISA system by using recombinant protein. Recombinant protein of LEDGF/p75 was purchased from Protein One (Bethesda, MD, USA). We developed an ELISA system to detect anti-LEDGF/p75 by coating this recombinant protein. 226 sera of AIFS patients (including 36 CFS patients) were applied to this ELISA assay and Western immunoblot, and it was revealed that anti-Sa-positive sera defined by WB and sera positive for anti-LEDGF/p75 on ELISA were identical. Moreover, reactivities of anti-Sa on WB were inhibited by pre-incubating with recombinant LEDGF/p75, and eluted antibodies from the nitrocellulose membrane could react on the ELISA. These results confirm that the Sa antigen is LEDGF/p75. The ELISA assay using recombinant LEDGF/p75 could be a promising tool for measuring anti-Sa and consequently for diagnosing CFS. PMID:19657776

  10. Association of antinuclear and antinucleolar antibodies in progressive systemic sclerosis.

    PubMed Central

    Bernstein, R M; Steigerwald, J C; Tan, E M

    1982-01-01

    Antinuclear and/or antinucleolar antibodies were demonstrated in the sera of 74 of 76 patients (97%) with progressive systemic sclerosis, using tissue culture cells (HEp-2) as substrate in the indirect immunofluorescent method. Six patterns of nuclear staining and three nucleolar patterns were recognized. The nuclear patterns were centromere, fine speckles, coarse speckles, diffusely grainy, homogeneous and nuclear dots. The nucleolar patterns were speckled, homogeneous and clumpy. The results of digestion studies with ribonuclease, deoxyribonuclease and trypsin suggested that the nuclear antigens are proteins, some of which may be associated with chromatin. The nucleolar antigens appeared to be nucleic acid in nature. Certain characteristic serologic and clinical features associated with staining patterns were observed. The diffusely grainy pattern was seen only in sera containing antibody to Scl-70 antigen. Centromere staining was confirmed to be highly selective for the CREST (Calcinosis, Raynaud's phenomenon, esophageal involvement, sclerodactyly and telangiectasis) variant of progressive systemic sclerosis with rheumatoid factor titres higher in these patients with anti-centromere antibodies. PMID:7044633

  11. Precise quantitation of antinuclear antibodies on HEp-2 cells without the need for serial dilution.

    PubMed Central

    Hollingsworth, P N; Dawkins, R I; Peter, J B

    1996-01-01

    Using HEp-2 cells as a substrate, we developed a method to quantitate antinuclear antibodies (ANA) by comparing the green fluorescence intensity of unknown samples with that of calibrated standards. Intensity was then converted to international units per milliliter by reference to a standard curve. This method is accurate and precise around the cutoff for positively (5 to 10 IU/ml) and therefore provides a reliable screening test for active, untreated systemic lupus erythematosus. Furthermore, the method can identify sera likely to contain autoantibodies commonly detected in ANA-positive sera (SS-A, SS-B, Sm, small nuclear ribonucleoprotein, Scl-70, and double-stranded DNA). PMID:8807199

  12. Human peripheral blood monocytes display surface antigens recognized by monoclonal antinuclear antibodies

    SciTech Connect

    Holers, V.M.; Kotzin, B.L.

    1985-09-01

    The authors used monoclonal anti-nuclear autoantibodies and indirect immunofluorescence to examine normal human peripheral blood mononuclear leukocytes for the presence of cell surface nuclear antigens. Only one monoclonal anti-histone antibody (MH-2) was found to bind to freshly isolated PBL, staining approximately 10% of large cells. However, after cells were placed into culture for 16-24 h, a high percentage (up to 60%) of large-sized cells were recognized by an anti-DNA (BWD-1) and several different antihistone monoclonal antibodies (BWH-1, MH-1, and MH-2). These antibodies recognize separate antigenic determinants on chromatin and histones extracted from chromatin. The histone antigen-positive cells were viable, and the monoclonal antibodies could be shown to be binding to the cell surface and not to the nucleus. Using monoclonal antibodies specific for monocytes and T cells, and complement-mediated cytotoxicity, the cells bearing histone antigens were shown to be primarily monocytes. The appearance of histone and DNA antigen-positive cells was nearly completely inhibited by the addition of low concentrations of cycloheximide at initiation of the cultures. In contrast, little effect on the percentage of positive cells was detected if cells were exposed to high doses of gamma irradiation before culture. These data further support the existence of cell surface nuclear antigens on selected cell subsets, which may provide insight into the immunopathogenesis of systemic lupus erythematosus and related autoimmune diseases.

  13. Evaluation of circulating immune complexes and antinuclear antibodies in Japanese patients with leprosy

    Microsoft Academic Search

    F. Furukawa; K. Sekita; Y. Hamashima; M. Ozaki; S. Imamura

    1983-01-01

    In 79 patients with leprosy a significant increase of anti-extractable nuclear antigen (ENA) antibodies and circulating immune complexes (CIC) was found. No correlation between CIC and anti-ENA antibodies was demonstrable. Since such a correlation is known from antinuclear antibodies and CIC in patients with systemic lupus erythematosus, it appears likely that anti-ENA antibodies do not play a causative role in

  14. Anti-Nuclear Antibodies in Daily Clinical Practice: Prevalence in Primary, Secondary, and Tertiary Care

    PubMed Central

    Avery, Thomas Y.; van de Cruys, Mart; Austen, Jos; Stals, Frans; Damoiseaux, Jan G. M. C.

    2014-01-01

    For the diagnosis of systemic autoimmune rheumatic diseases (SARD), patients are screened for anti-nuclear antibodies (ANA). ANA, as assessed by indirect immunofluorescence (IIF), have a poor specificity. This hampers interpretation of positive results in clinical settings with low pretest probability of SARD. We hypothesized that the utility of positive ANA IIF results increases from primary to tertiary care. We retrospectively determined ANA, anti-ENA, and anti-dsDNA antibody prevalence in patient cohorts from primary (n = 1453), secondary (n = 1621), and tertiary (n = 1168) care settings. Results reveal that from primary care to tertiary care, ANA prevalence increases (6.2, 10.8, and 16.0%, resp.). Moreover, in primary care low titres (70% versus 51% and 52% in secondary and tertiary care, resp.) are more frequent and anti-ENA/dsDNA reactivities are less prevalent (21% versus 39% in secondary care). Typically, in tertiary care the prevalence of anti-ENA/dsDNA reactivities (21%) is lower than expected. From this descriptive study we conclude that positive ANA IIF results are more prone to false interpretation in clinical settings with low pretest probabilities for SARD, as in primary care. Whether alternative approaches, that is, immunoadsorption of anti-DFS70 antibodies or implementation of anti-ENA screen assays, perform better, needs to be determined. PMID:24741596

  15. Induction of anti-nuclear antibodies in mice orally exposed to cadmium at low concentrations.

    PubMed Central

    Ohsawa, M; Takahashi, K; Otsuka, F

    1988-01-01

    Anti-nuclear antibodies (ANA) in serum were detected in male ICR mice fed drinking water containing 3, 30 and 300 ppm Cd as CdCl2 for 10 weeks. In response to Cd exposure, ICR mice developed ANA of the IgG class giving nuclear patterns moderately stained by immunofluorescence or immunoenzyme method. Positive immunofluorescence staining of ANA was obtained in 50, 89 and 90% of ICR mice exposed to 3, 30 and 300 ppm Cd, respectively. Their spleen cells also showed an enhancement of antibody forming response to sheep red blood cells (SRBC) without the SRBC priming. When mice were primed with SRBC after exposure to Cd, however, a significant suppression of the antibody forming response was observed in mice fed 300 ppm Cd but not in those fed 3 ppm Cd. No significant differences in delayed-type hypersensitivity reaction to SRBC were observed between Cd-fed and control animals. Inbred BALB/c mice were less susceptible to the induction of ANA by Cd, as induced only in 300 ppm Cd-fed mice. Thus environmental exposure to Cd can induce ANA in ICR mice with a high susceptibility, presumably accompanied with a non-specific stimulation of antibody formation. Images Fig. 1 PMID:3048815

  16. Induction of antinuclear antibodies in patients with rheumatoid arthritis receiving treatment with human recombinant interferon gamma.

    PubMed Central

    Seitz, M; Franke, M; Kirchner, H

    1988-01-01

    Of six patients with rheumatoid arthritis (RA) treated with human recombinant interferon gamma for two to eight months, three developed antinuclear antibodies (ANAs). This was accompanied by a simultaneous clinical exacerbation of the disease activity. In this study both anti-inflammatory and immunostimulatory effects of human recombinant interferon gamma in patients with RA were observed. PMID:3137901

  17. Editorial Overview: Antinuclear Antibody and Extractable Nuclear Antigen-Related Diseases

    Microsoft Academic Search

    Falk Hiepe; Thomas Dörner; Gerd-Rüdiger Burmester

    2000-01-01

    In 1948, the observation of the LE cell phenomenon in a patient with systemic lupus erythematosus (SLE) began the discovery of a broad variety of autoantibodies directed to nuclear antigens called antinuclear antibodies (ANA). Nowadays, different ANA serve as important diagnostic parameters for differentiating most of the connective tissue diseases, such as SLE, neonatal lupus syndromes, Sjögren’s syndrome, scleroderma, autoimmune

  18. Cold agglutinin-induced haemolysis in association with antinuclear antibody-negative SLE

    PubMed Central

    Chaubey, Vinod K; Chhabra, Lovely

    2013-01-01

    Systemic lupus erythematosus (SLE) is a chronic relapsing autoimmune disease associated with several autoantibodies targeted to nuclear and cytoplasmic antigens. Serum antinuclear antibody (ANA) is considered an important diagnostic marker of SLE. However, 2–3% of patients with typical clinical picture of SLE may have persistently negative ANA tests. Autoimmune haemolytic anaemia (AIHA) in SLE is usually mediated by warm IgG anti-erythrocyte antibodies. Our report describes a female patient who presented with clinical manifestations of SLE including photosensitivity, joint pains and AIHA. Further workup revealed high cold IgM agglutinin titres. A comprehensive workup for infectious aetiologies was negative. Autoimmune studies revealed negative ANA, but positive anti-double-stranded DNA and antiphospholipid antibodies. Lymphoproliferative disorder was excluded by imaging studies. Initial treatment with steroids proved of little benefit; however, rituximab resulted in significant clinical improvement. To the best of our knowledge, this is perhaps the first report of ANA-negative SLE presenting with cold AIHA. PMID:23761498

  19. The Prevalence of Antinuclear Antibodies in the General Population of China: A Cross-Sectional Study

    PubMed Central

    Guo, Ya-Ping; Wang, Chun-Guang; Liu, Xin; Huang, Yi-Qian; Guo, De-Li; Jing, Xing-Zhuo; Yuan, Chun-Gang; Yang, Song; Liu, Jin-Mei; Han, Meng-Si; Li, Hong-Xing

    2014-01-01

    Background The incidence of autoimmune diseases such as systemic lupus erythematosus, rheumatoid arthritis, and primary biliary cirrhosis has increased significantly in China. Information about the susceptibility or potential of autoimmune diseases in the general population is lacking. Objective To explore the prevalence of antinuclear antibody (ANA) and its specificities in the general population in China. Methods Twenty thousand nine hundred seventy sera samples were taken from the physical examination center in Baoding, China. Indirect immunofluorescence and line immunoassays were used to detect ANA and its specificities, respectively. Results Samples from females had a higher prevalence of ANA than samples from males (?2 = 278.55; P < 0.01). For both sexes, the prevalence of ANA positively correlated with age and there were significant differences among different age groups at 10-year intervals, except the 80 years group (P < 0.05). One thousand two hundred forty-three ANA-positive samples were further analyzed with line immunoassays. There was a significant difference among age groups and between sex groups in terms of the specific autoantibodies (P < 0.01). The autoantibodies with the top-3 positive frequencies were anti-Ro-52, anti-M2, and anti-SSA. Conclusions There was a high prevalence of ANA positivity in the general Chinese population that seemed to be influenced by sex and age and correlated with specific autoantibodies. PMID:25473438

  20. 21 CFR 866.5100 - Antinuclear antibody immunological test system.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ...such as ribonucleic acid, deoxyribonucleic acid, or nuclear proteins). The measurements aid in the diagnosis of systemic lupus erythematosus (a multisystem autoimmune disease in which antibodies attack the victim's own tissues), hepatitis...

  1. 21 CFR 866.5100 - Antinuclear antibody immunological test system.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...such as ribonucleic acid, deoxyribonucleic acid, or nuclear proteins). The measurements aid in the diagnosis of systemic lupus erythematosus (a multisystem autoimmune disease in which antibodies attack the victim's own tissues), hepatitis...

  2. 21 CFR 866.5100 - Antinuclear antibody immunological test system.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ...such as ribonucleic acid, deoxyribonucleic acid, or nuclear proteins). The measurements aid in the diagnosis of systemic lupus erythematosus (a multisystem autoimmune disease in which antibodies attack the victim's own tissues), hepatitis (a...

  3. 21 CFR 866.5100 - Antinuclear antibody immunological test system.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ...such as ribonucleic acid, deoxyribonucleic acid, or nuclear proteins). The measurements aid in the diagnosis of systemic lupus erythematosus (a multisystem autoimmune disease in which antibodies attack the victim's own tissues), hepatitis (a...

  4. 21 CFR 866.5100 - Antinuclear antibody immunological test system.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ...such as ribonucleic acid, deoxyribonucleic acid, or nuclear proteins). The measurements aid in the diagnosis of systemic lupus erythematosus (a multisystem autoimmune disease in which antibodies attack the victim's own tissues), hepatitis...

  5. Negatively charged polymers protect antinuclear antibody against inactivation by acylating agents.

    PubMed

    Samokhin, G P; Mongayt, D A; Iakoubov, L Z; Levchenko, T S; Torchilin, V P

    2001-05-15

    For many practical applications, monoclonal antibodies must be chemically modified without any significant loss in their immunoreactivity. In some situations, however, the amino acid residue crucial for antibody activity may be highly reactive toward the modifying agent, which results in antibody inactivation. The method to prevent inactivation of a modification-sensitive antinuclear monoclonal antibody by acylating agents was developed. The method is based on the hypothesis that a highly reactive amino group exists within, or in the vicinity of, the binding site of the antibody, providing crucial interaction with negatively charged moieties of DNA. It has been shown that negatively charged polymers, such as dextran sulfate or heparin, may provide temporary protection, presumably interacting noncovalently with this amino group and thus masking it. The protecting molecule can be removed later by chromatography on a protein A column, thus regenerating modified but not inactivated antibody in the free form for use in subsequent applications. In particular, we have modified antibody 2C5 with a chelating agent, diethylenetriaminepentaacetic acid (DTPA) without the loss of activity. Modified antibody was labeled with radioactive isotope, (111)In, via chelation by antibody-attached DTPA. The labeled antibody was shown to demonstrate the same specificity of binding to nucleosomes as the nonmodified antibody, so it may be used in immunoscintigraphy or biodistribution studies. The method might be useful for the modification of other modification-sensitive antibodies with other acylating chemicals, such as crosslinking agents or biotin derivatives. PMID:11355857

  6. [Baby hamster kidney cells as antigen for demonstration of antinuclear antibodies (author's transl)].

    PubMed

    Pernice, W; Scherer, R; Lüben, G; Sodomann, C P; Sedlacek, H H

    1978-09-15

    Baby hamster kidney cells fixed in acetone on glass slides were used as antigen for demonstration of antinuclear antibodies. Where certain storage conditions were observed (drying agent, 4 degrees C) they have kept for 12 months up to now. As regards specificity, sensitivity, reproducibility, and differentiation of fluorescent types the baby hamster kidney cell test appears superior to other immunofluorescence methods used (chicken erythrocytes, rat liver sections, and crithidiae). These results were obtained in 73 sera from patients with disseminated lupus erythematodes, drug-induced lupus erythematodes, discoid erythematodes, allergic vasculitis, progressive scleroderma, dermatomyositis, and 36 control sera. PMID:308447

  7. Prevalence of disease-specific antinuclear antibodies in general population: estimates from annual physical examinations of residents of a small town over a 5-year period

    Microsoft Academic Search

    Nobuhide Hayashi; Masahiro Koshiba; Kunihiro Nishimura; Daisuke Sugiyama; Tomoko Nakamura; Sahoko Morinobu; Seiji Kawano; Shunichi Kumagai

    2008-01-01

    The aim of this study was to investigate the types and prevalence of disease-specific antinuclear antibodies (ANAs) and their\\u000a relationship to rheumatic diseases in the general Japanese population. An immunofluorescence (IF) method was used for the\\u000a first screening of ANA levels in serum samples obtained from 2181 residents of a small Japanese town. Individuals positive\\u000a for IF-ANA were then further

  8. A bag of cells approach for antinuclear antibodies HEp-2 image classification.

    PubMed

    Wiliem, Arnold; Hobson, Peter; Minchin, Rodney F; Lovell, Brian C

    2015-06-01

    The antinuclear antibody (ANA) test via indirect immunofluorescence applied on Human Epithelial type 2 (HEp-2) cells is a pathology test commonly used to identify connective tissue diseases (CTDs). Despite its effectiveness, the test is still considered labor intensive and time consuming. Applying image-based computer aided diagnosis (CAD) systems is one of the possible ways to address these issues. Ideally, a CAD system should be able to classify ANA HEp-2 images taken by a camera fitted to a fluorescence microscope. Unfortunately, most prior works have primarily focused on the HEp-2 cell image classification problem which is one of the early essential steps in the system pipeline. In this work we directly tackle the specimen image classification problem. We aim to develop a system that can be easily scaled and has competitive accuracy. ANA HEp-2 images or ANA images are generally comprised of a number of cells. Patterns exhibiting in the cells are then used to make inference on the ANA image pattern. To that end, we adapted a popular approach for general image classification problems, namely a bag of visual words approach. Each specimen is considered as a visual document containing visual vocabularies represented by its cells. A specimen image is then represented by a histogram of visual vocabulary occurrences. We name this approach as the Bag of Cells approach. We studied the performance of the proposed approach on a set of images taken from 262 ANA positive patient sera. The results show the proposed approach has competitive performance compared to the recent state-of-the-art approaches. Our proposal can also be expanded to other tests involving examining patterns of human cells to make inferences. © 2014 International Society for Advancement of Cytometry. PMID:25492545

  9. Immune-Mediated Fever in the Dog. Occurrence of Antinuclear Antibodies, Rheumatoid Factor, Tumor Necrosis Factor and Interleukin-6 in Serum

    PubMed Central

    Bohnhorst, Øvrebø; Hanssen, I; Moen, Torolf

    2002-01-01

    Contents of antinuclear antibodies (ANA), rheumatoid factor (RF), tumor necrosis factor (TNF-?) and interleukin-6 (IL-6) were measured in serum from 20 dogs with immune-mediated fever. Seven out of 20 patients were ANA positive, 1 out of 20 was positive to antibodies against extractable nuclear antigens (ENA), 1 out of 20 was positive to antibodies against deoxynucleoproteins (DNP), 2 out of 13 were RF positive and none out of 20 patients had antibodies against native DNA in the serum. TNF-? was not detected in any serum of 15 dogs with immune-mediated fever, while 10 out of 13 presented with elevated IL-6. The results varied between patients, but the IL-6 level was high in most of them. This indicate a role for IL-6 in the pathogenesis of immune-mediated fever in most cases. PMID:12564546

  10. Cronkhite-Canada syndrome showing elevated levels of antinuclear and anticentromere antibody.

    PubMed

    Ota, Seisuke; Kasahara, Akinori; Tada, Shoko; Tanaka, Takehiro; Umena, Sachio; Fukatsu, Haruka; Noguchi, Toshio; Matsumura, Tadashi

    2015-02-01

    A 56-year-old female initially visited an otorhinolaryngologist because of an impaired sense of taste in September, 2010 and was referred to our facility in October, 2010. She was diagnosed with Basedow's disease for which she underwent subtotal thyroidectomy in 1984 and arthritis involving multiple joints, primarily affecting her hands. In addition, the anticentromere antibody (ACA) level was markedly high. On physical examination, alopecia as well as hyperpigmentation of the dorsum of the hands and back was observed. Dystrophic changes of the fingernails and a bilateral thumb abduction deformity were observed. Antinuclear antibodies were elevated. Gastrointestinal endoscopy and colonoscopy revealed the mucosa carpeted with strawberry-like polypoid lesions. Histopathological examination of the biopsied specimen of the stomach revealed a corkscrew-like appearance. Thus, the patient was diagnosed with Cronkhite-Canada syndrome (CCS). She admitted to our hospital in November, 2010. Oral prednisolone was administered with success. In July, 2012, her antimitochondrial M2 antibody level was elevated. To the best of our knowledge, the present case is the first patient with CCS, a history of Basedow's disease, and elevated levels of ACA and antimitochondrial M2 antibody. We consider the present case suggests CCS could be caused by immunological abnormality. PMID:25518819

  11. Rheumatoid factor, anti-nuclear antibody in ischemic heart disease: Acute versus chronic patients

    PubMed Central

    Sedaghat, Akram; Sadeghi, Masoumeh; Heidari, Ramin; Sistani, Efat; Bayanfar, Zahra

    2014-01-01

    BACKGROUND Immunopathological and inflammatory processes play important roles in the initiation and development of ischemic heart disease. Hence, this study aimed to evaluate the relationship between serum levels rheumatoid factor (RF) and anti-nuclear antibodies (ANA) and severity of coronary stenotic lesions. METHODS Totally 140 patients with acute coronary syndrome (ACS) (n = 70) and chronic stable angina (CSA) (n = 70) that undergoing coronary angiography were enrolled in this study. ANA by the enzyme-linked immunosorbent assay (ELISA) and serum level of RF was measured by latex method. The severity of coronary stenotic lesions calculated by Gensini score. To analyze the correlations of ANA and RF to Gensini score Pearson correlation test was used. To adjust the effect of age and other confounder factors such hypertension, diabetes, hyperlipidemia and smoking multiple linear regression was used. RESULTS The mean serum levels of RF and ANA in CSA group were significantly higher than ACS group after adjusting for the confounder factors (P < 0.050 for ANA). Serum levels of ANA significantly correlated with severity of coronary stenotic lesions calculated by Gensini score (r = 0.40 and P < 0.050). After adjusting confounders, multiple linear regression analysis showed ANA remained independently associated with Gensini scores in ACS group (B = 0.505, P < 0.001). CONCLUSION Higher serum levels of ANA may be considered as independent risk factors for ACS. PMID:25815020

  12. Anti-telomere antibodies in systemic lupus erythematosus (SLE): a comparison with five antinuclear antibody assays in 430 patients with SLE and other rheumatic diseases

    PubMed Central

    Salonen, E; Miettinen, A; Walle, T; Koskenmies, S; Kere, J; Julkunen, H

    2004-01-01

    Objective: To investigate the prevalence and diagnostic significance of antibodies against telomeric DNA in systemic lupus erythematosus (SLE) and other autoimmune rheumatic diseases, and to make comparisons with five conventional anti-DNA or anti-nuclear antibody (ANA) assays. Methods: Antibodies to telomeres, which are highly repetitive sequences of DNA (TTAGGG/CCCTAA) at the end of eukaryotic chromosomes, were measured by an enzyme linked immunosorbent assay (ELISA) in 305 patients with SLE and 125 patients with other autoimmune rheumatic diseases (78 rheumatoid arthritis, 32 primary Sjögren's syndrome, eight mixed connective tissue disease, seven miscellaneous rheumatic diseases). Other assays used were two commercial ELISA assays for anti-dsDNA using calf thymus as antigen, Crithidialuciliae immunofluorescence, and radioimmunoassay (RIA) for anti-dsDNA and immunofluorescence using Hep-2 cells for ANA. Results: The prevalence of anti-telomere in SLE was 60%, v 5% in rheumatoid arthritis and 18% in other autoimmune rheumatic diseases. Specificity of anti-telomere for SLE was 91%; positive and negative predictive values were 95% and 46%, respectively. For anti-dsDNA by two ELISA assays using calf thymus as antigen, sensitivities were 69% and 29% and specificities 66% and 96%, respectively. Other anti-dsDNA assays had low sensitivities (RIA 43%, Crithidia immunofluorescence 13%). The association of anti-telomere with a history of nephritis in patients with SLE was stronger (p = 0.005) than by any other assay (p = 0.006–0.999). The correlations between the different assays were good (p<0.001 for all comparisons). Conclusions: The new ELISA for anti-telomere antibodies using standardised human dsDNA as antigen is a sensitive and highly specific test for SLE. PMID:15361381

  13. Anti-SSA/Ro antibody determination by enzyme-linked immunosorbent assay as a supplement to standard immunofluorescence in antinuclear antibody screening.

    PubMed

    Blomberg, S; Ronnblom, L; Wallgren, A C; Nilsson, B; Karlsson-Parra, A

    2000-06-01

    The aim of this study was to investigate the frequency and possible clinical relevance of SSA/Ro antibodies, as determined by enzyme-linked immunosorbent assay (ELISA), in patient sera not exhibiting a concomitant positive reaction by the standard immunofluorescence (IF) test using HEP-2 cells as substrate. SSA/Ro reactivity, as shown by ELISA, was found in 285 (7%) of 4025 serum samples consecutively remitted for antinuclear antibody (ANA) screening. Seventy-five of these serum samples (26%), derived from 64 patients, were negative by the IF-ANA screening test. Serum samples from all 64 patients exhibiting SSA/Ro reactivity by ELISA without concomitant positivity by IF-ANA were further investigated by IF using transfected HEP-2 cells hyperexpressing the 60,000 MW SSA/Ro antigen (HEP-2000(R)) and by immunodiffusion (ID) and Western blot. In 55 of these 64 patients, SSA/Ro reactivity could be verified by one or more of the other techniques investigated. Twelve of these patients fulfilled four or more American College of Rheumatology (ACR) criteria for systemic lupus erythematosus (SLE) and another five patients exhibited a histologically confirmed cutaneous lupus erythematosus (LE). In four of the 12 IF-ANA-negative patients with a diagnosis of SLE, the SSA/Ro reactivity was only detectable by ELISA and Western blot. In conclusion, the use of a sensitive ELISA assay could provide a clinically important supplement to the routine ANA screening by IF, which does not detect certain anti-SSA/Ro-containing sera among patients with relevant autoimmune diagnoses. Detection of anti-SSA/Ro antibodies, however, does not alone signify cutaneous LE or SLE but adds weight to these diagnoses that should rely heavily on other clinical information. PMID:10849373

  14. Prevalence of disease-specific antinuclear antibodies in general population: estimates from annual physical examinations of residents of a small town over a 5-year period.

    PubMed

    Hayashi, Nobuhide; Koshiba, Masahiro; Nishimura, Kunihiro; Sugiyama, Daisuke; Nakamura, Tomoko; Morinobu, Sahoko; Kawano, Seiji; Kumagai, Shunichi

    2008-01-01

    The aim of this study was to investigate the types and prevalence of disease-specific antinuclear antibodies (ANAs) and their relationship to rheumatic diseases in the general Japanese population. An immunofluorescence (IF) method was used for the first screening of ANA levels in serum samples obtained from 2181 residents of a small Japanese town. Individuals positive for IF-ANA were then further tested for disease-specific ANAs using eight enzyme immunoassays. Physical status and the presence of illness were determined by means of questionnaires and medical examinations. Based on the result of the IF-ANA assay, the rates of positive samples at 1:40 and 1:160 dilutions were 26.0 and 9.5%, respectively, with females have significantly higher positivity rates than males (P < 0.0001). Among 566 IF-ANA-positive individuals, 100 individuals were found to have 114 disease-specific ANAs. Anti-SSA/Ro, anti-centromere, and anti-U1RNP antibodies were detected in 58, 30, and 11 individuals, respectively, but anti-Sm, anti-Scl-70, and anti-Jo-1 antibodies were undetectable. Questionnaires and medical examinations revealed that among 60 disease-specific ANA-positive individuals that were available for testing, six had Sjögren's syndrome (SS), five were suspected of having SS, and five had rheumatoid arthritis. Surprisingly, 34 (57%) of the disease-specific ANA-positive individuals were clinically healthy. Anti-SSA/Ro, anti-centromere, and anti-U1RNP antibodies were quite frequent among clinically healthy Japanese subjects, although anti-Sm, anti-Scl-70, and anti-Jo-1 antibodies were not. Of the 60 individuals who tested positive for disease-specific ANAs, 30% (18/60) actually manifested systemic rheumatic diseases, while 50% showed no detectable signs or symptoms of rheumatic diseases. PMID:18283522

  15. TLR Tolerance Reduces IFN-Alpha Production Despite Plasmacytoid Dendritic Cell Expansion and Anti-Nuclear Antibodies in NZB Bicongenic Mice

    Microsoft Academic Search

    Evelyn Pau; Yui-Ho Cheung; Christina Loh; Ginette Lajoie; Joan E. Wither

    2012-01-01

    Genetic loci on New Zealand Black (NZB) chromosomes 1 and 13 play a significant role in the development of lupus-like autoimmune disease. We have previously shown that C57BL\\/6 (B6) congenic mice with homozygous NZB chromosome 1 (B6.NZBc1) or 13 (B6.NZBc13) intervals develop anti-nuclear antibodies and mild glomerulonephritis (GN), together with increased T and B cell activation. Here, we produced B6.NZBc1c13

  16. Role of Nucleic Acid–Sensing TLRs in Diverse Autoantibody Specificities and Anti-nuclear Antibody–Producing B Cells

    PubMed Central

    Koh, Yi Ting; Scatizzi, John C.; Gahan, Jennifer D.; Lawson, Brian R.; Baccala, Roberto; Pollard, K. Michael; Beutler, Bruce A.; Theofilopoulos, Argyrios N.; Kono, Dwight H.

    2013-01-01

    Nucleic acid (NA)–sensing TLRs (NA-TLRs) promote the induction of anti-nuclear Abs in systemic lupus erythematosus. However, the extent to which other nonnuclear pathogenic autoantibody specificities that occur in lupus and independently in other autoimmune diseases depend on NA-TLRs, and which immune cells require NA-TLRs in systemic autoimmunity, remains to be determined. Using Unc93b13d lupus-prone mice that lack NA-TLR signaling, we found that all pathogenic nonnuclear auto-antibody specificities examined, even anti-RBC, required NA-TLRs. Furthermore, we document that NA-TLRs in B cells were required for the development of antichromatin and rheumatoid factor. These findings support a unifying NA-TLR–mediated mechanism of autoantibody production that has both pathophysiological and therapeutic implications for systemic lupus erythematosus and several other humoral-mediated autoimmune diseases. In particular, our findings suggest that targeting of NA-TLR signaling in B cells alone would be sufficient to specifically block production of a broad diversity of autoantibodies. PMID:23589617

  17. Outcome of asbestos exposure (lung fibrosis and antinuclear antibodies) with respect to skin reactivity: an 8-year longitudinal study

    SciTech Connect

    Lange, A.; Garncarek, D.; Tomeczko, J.; Ciechanowski, G.; Bisikiewicz, R.

    1986-10-01

    Two hundred seventy asbestos workers were examined during an 8-year period. During this time five consecutive surveys were completed. Skin tests with streptokinase-streptodornase (SK-SD), tuberculin (PPD), and phytohemagglutinin (PHA) were performed in the middle of this period. The results of these tests were related to X-ray chest film results and the appearance of antinuclear antibodies (ANA). In all surveys, except the first, X-ray films with small irregular opacities with a profusion greater than or equal to1/1 belonged more frequently to asbestos workers who did not respond to SK-SD or PHA. Thirty-one cases of asbestosis were diagnosed at that time, 23 of them became asbestotic after the skin tests were performed. Asbestotic cases contributed more frequently to the group with energy as compared to asbestos workers lacking asbestosis. Furthermore, asbestosis was correlated with lack of response to second strength of SK-SD in males and PHA in both sexes. Lack of response to these activators was predictive of asbestosis. Asbestos workers with ANA frequently displayed a lack of response to the first strength of SK-SD, PPD, and PHA. This was partly due to the presence of asbestotic cases in the group. However, low responders to all these activators were found frequently in the group with ANA independent of the presence of asbestosis.

  18. Development of antinuclear antibodies and a genetic linkage in pigs infected with porcine circovirus type 2

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Objectives. Prominent nuclear immunohistochemical staining of a PCV-2 free porcine kidney cell line (PK-15) was detected with a rabbit polyclonal antibody produced against a conserved PCV2 Rep-protein peptide. This unexpected finding led us to retrospectively test sera from gnotobiotic pigs for the ...

  19. ANA (Antinuclear Antibody Test)

    MedlinePLUS

    ... one of the tests to help diagnose systemic lupus erythematosus (SLE) . ANA are a group of autoantibodies ... in one or more joints Red rash (for lupus , one resembling a butterfly across the nose and ...

  20. Antinuclear antibody panel

    MedlinePLUS

    ... have signs of an autoimmune disorder, particularly systemic lupus erythematosus . This test may be done if you ... ANA does not confirm a diagnosis of systemic lupus erythematosis (SLE). However, a lack of ANA makes ...

  1. Prevalence of antinuclear antibodies in 3 groups of healthy individuals: blood donors, hospital personnel, and relatives of patients with autoimmune diseases.

    PubMed

    Marin, Guadalupe G; Cardiel, Mario H; Cornejo, Horacio; Viveros, Martha E

    2009-10-01

    Antinuclear antibodies (ANA) are frequently found in healthy populations. To define the prevalence, pattern, and titer of ANA in different groups of the healthy Mexican population, we studied 304 individuals, classified into 3 groups: 104 blood donors, 100 hospital personnel working at The State General Hospital, which included doctors, laboratory technicians, and nurses; and 100 relatives of patient diagnosed either with systemic lupus erythematosus or rheumatoid arthritis, all of them apparently healthy at the time of study. We determined ANA using immunofluorescence microscopy performed on HEp-2 cells. Fluorescence was detected in 165 serum samples (54.3%). The most frequent pattern was the speckled (50.3%). The most frequent dilution was 1:40 (35.4%), followed by 1:80 (13.4%), 1:160 (3.2%), and 1:320 (1.3%).Regarding the results by study group, we found a trend toward higher ANA levels in group 2 (hospital personnel), compared with group 1 (blood donors) and group 3 (relatives of patients), a trend also reflected by the increasing frequency of serum titers of 1:80 and higher (P = 0.074). According to occupation, medical doctors showed a higher incidence of speckled pattern when compared with other occupations (P = 0.022). Medical doctors (n = 75) showed also higher titers of this particular pattern (P = 0.03). In group 3, relatives of patients with systemic lupus erythematosus showed the speckled pattern more frequently than relatives of patients with rheumatoid arthritis, in low titers (P = 0.017). We suggest that ANA tests showing speckled pattern should be at a 1:160 titer or higher to be considered positive; other patterns such as homogeneous, peripheral, or centromeric might be considered positive even at low titers (Positive ANA finding in the absence of physical signs and symptoms has limited diagnostic utility and should always be interpreted by a rheumatologist, in the context of clinical symptoms and results of laboratory tests for specific autoantibodies. Populations such as doctors and relatives of patients with autoimmune disease tend to presents increased ANA titers. PMID:20009966

  2. Mercury exposure, malaria, and serum antinuclear/antinucleolar antibodies in amazon populations in Brazil: a cross-sectional study

    PubMed Central

    Silva, Ines A; Nyland, Jennifer F; Gorman, Andrew; Perisse, Andre; Ventura, Ana Maria; Santos, Elizabeth CO; de Souza, Jose M; Burek, CL; Rose, Noel R; Silbergeld, Ellen K

    2004-01-01

    Background Mercury is an immunotoxic metal that induces autoimmune disease in rodents. Highly susceptible mouse strains such as SJL/N, A.SW, B10.S (H-2s) develop multiple autoimmune manifestations after exposure to inorganic mercury, including lymphoproliferation, elevated levels of autoantibodies, overproduction of IgG and IgE, and circulating immune complexes in kidney and vasculature. A few studies have examined relationships between mercury exposures and adverse immunological reactions in humans, but there is little evidence of mercury-associated autoimmunity in humans. Methods To test the immunotoxic effects of mercury in humans, we studied communities in Amazonian Brazil with well-characterized exposures to mercury. Information was collected on diet, mercury exposures, demographic data, and medical history. Antinuclear and antinucleolar autoantibodies (ANA and ANoA) were measured by indirect immunofluorescence. Anti-fibrillarin autoantibodies (AFA) were measured by immunoblotting. Results In a gold mining site, there was a high prevalence of ANA and ANoA: 40.8% with detectable ANoA at ?1:10 serum dilution, and 54.1% with detectable ANA (of which 15% had also detectable ANoA). In a riverine town, where the population is exposed to methylmercury by fish consumption, both prevalence and levels of autoantibodies were lower: 18% with detectable ANoA and 10.7% with detectable ANA. In a reference site with lower mercury exposures, both prevalence and levels of autoantibodies were much lower: only 2.0% detectable ANoA, and only 7.1% with detectable ANA. In the gold mining population, we also examined serum for AFA in those subjects with detectable ANoA (?1:10). There was no evidence for mercury induction of this autoantibody. Conclusions This is the first study to report immunologic changes, indicative of autoimmune dysfunction in persons exposed to mercury, which may also reflect interactions with infectious disease and other factors. PMID:15522122

  3. A retrospective study on IVF/ICSI outcome in patients with anti-nuclear antibodies: the effects of prednisone plus low-dose aspirin adjuvant treatment

    PubMed Central

    2013-01-01

    Background Anti-nuclear antibodies (ANA) are suspected of having relevance to adverse reproductive events. Methods This study aims to investigate the potential effect of ANA on IVF/ICSI outcome and the therapeutic role of prednisone plus low-dose aspirin (P + A) adjuvant treatment in ANA + patients. The first IVF/ICSI cycles without P + A of sixty-six ANA + women were enrolled as the ANA + group, and the 233 first IVF/ICSI cycles of matched ANA- women served as the ANA- group. The ANA + group was divided into the Titre??1:320 subgroup. Twenty-one ANA + women with adverse outcomes in their first cycles (ANA + cycles without P + A) received P + A adjuvant treatment for three months before the second IVF/ICSI cycle (ANA + cycles with P + A). The clinical characteristics and the IVF/ICSI outcomes were compared, respectively, between 1) the ANA + group and the ANA- group, 2) the Titre??1:320 subgroup, and 3) the ANA + cycles without P + A and the ANA + cycles with P + A. Results No significant differences were observed between each of the two-group pairs in the clinical characteristics. The ANA?+?group exhibited significantly lower MII oocytes rate, normal fertilisation, pregnancy and implantation rates, as well as remarkably higher abnormal fertilisation and early miscarriage rates. The Titre??1:320 subgroup. After the P + A adjuvant treatment, the number of two pro-nuclei, perfect embryos and available embryos, and the implantation rate increased significantly. Conclusions These observations suggest that ANA could exert a detrimental effect on IVF/ICSI outcome that might not be titre-dependent, and P + A adjuvant treatment could be useful for ANA + patients. This hypothesis should be verified in further prospective randomised studies. PMID:24093222

  4. Detection of Antiendothelial Cell Antibodies by an Enzyme-Linked Immunosorbent Assay Using Antigens from Cell Lysate: Minimal Interference with Antinuclear Antibodies and Rheumatoid Factors

    Microsoft Academic Search

    Christian Drouet; Marie-France Nissou; Denise Ponard; Josiane Arvieux; Chantal Dumestre-Perard; Philippe Gaudin; Bernard Imbert; Christian Massot; Francoise Sarrot-Reynauld

    2003-01-01

    Antiendothelial cell antibodies (AECAs), a heterogeneous group of autoantibodies, are associated with several diseases characterized by immune-mediated vascular damage (for re- views, see references 2 and 13), including systemic lupus ery- thematosus (19), systemic sclerosis (17), Wegener's granulo- matosis (6), and rheumatoid arthritis complicated by vasculitis (7). Even if they recognize poorly characterized targets, they may be valuable as markers

  5. Relation between antinuclear antibodies and the autoimmune rheumatic diseases and disease type and activity in systemic lupus erythematosus using a variety of cultured cell lines

    Microsoft Academic Search

    J Sequi; I Leigh; D A Isenberg

    1991-01-01

    Antinuclear activity was assessed in serum samples from a series of 40 patients with differing clinical subsets (including renal and neurological disease) of systemic lupus erythematosus (SLE) against a transformed keratinocyte line (SvK14)* and normal human keratinocytes. Paired serum samples were studied during disease activity and inactivity, and the effects of ultraviolet radiation on the availability of nuclear antigens in

  6. Pitfalls of formalin fixation for determination of antineutrophil cytoplasmic antibodies.

    PubMed Central

    Chowdhury, S M; Broomhead, V; Spickett, G P; Wilkinson, R

    1999-01-01

    Sera can produce nuclear or perinuclear immunofluorescence staining in neutrophils which may be caused by antibodies with differing antigenic specificities. These include perinuclear antineutrophil cytoplasmic antibodies (P-ANCA), granulocyte specific antinuclear antibody (GS-ANA), and antinuclear antibody (ANA). There is controversy over the value of formalin fixation of neutrophils in differentiating antibodies giving selective or preferential reaction with the nuclear or perinuclear area of neutrophils. In a comparative study of 77 sera, formalin fixation caused inconsistency, nonspecific effects, and false positivity owing to enhanced fluorescence. If formalin fixed neutrophils are used in the routine diagnostic laboratory, this will add confusion to the interpretation of the ANCA assay. PMID:10562820

  7. Development of connective tissue disease in patients presenting with Raynaud's phenomenon: a six year follow up with emphasis on the predictive value of antinuclear antibodies as detected by immunoblotting.

    PubMed Central

    Kallenberg, C G; Wouda, A A; Hoet, M H; van Venrooij, W J

    1988-01-01

    Eighty five patients referred because of Raynaud's phenomenon (RP) were followed up for six years. Every two years they were screened for signs and symptoms of connective tissue disease (CTD) according to a protocol, and serum was stored. Initially, 30 patients had primary RP, 16 had one symptom of CTD ('possible CTD'), 18 had two or more symptoms ('probable CTD'), and 21 had definite CTD (14 of whom had scleroderma). Most of the symptoms were related to scleroderma. There was an insidious progression to scleroderma or CRST syndrome (calcinosis, Raynaud's phenomenon, sclerodactyly, telangiectasia): 11 of 46 patients with primary RP or possible CTD developed probable scleroderma (two or more symptoms but not fulfilling all criteria), and seven of 13 patients with probable scleroderma developed definite scleroderma or CRST. The presence of distinct antinuclear antibodies (ANAs) as detected by immunoblotting in patients with primary RP and possible CTD at the start of the study was associated with the evolution of symptoms of CTD (chi 2 = 5.7, p less than 0.01). In patients initially with primary RP or possible CTD the antibody specificities of ANAs as determined by immunoblotting had prognostic value for the development of certain disease entities: anticentromere (CR-19) for CRST (sensitivity 60%, specificity 98%) and antitopoisomerase I (Scl-70 or Scl-86) for scleroderma or probable scleroderma (sensitivity 38%, specificity 100%). PMID:3261966

  8. Prevalence of symptoms of systemic lupus erythematosus (SLE) and of fluorescent antinuclear antibodies associated with chronic exposure to trichloroethylene and other chemicals in well water

    SciTech Connect

    Kilburn, K.H.; Warshaw, R.H. (Univ. of Southern California, Los Angeles (United States))

    1992-02-01

    Criteria for the recognition of systemic lupus erythematosus (SLE) were applied to 362 subjects exposed to trichloroethylene, trichloroethane, inorganic chromium, and other chemicals in water obtained from wells in an industrially contaminated aquifer in Tucson, Arizona. Their antinuclear autoantibodies were measured by fluorescence (FANA) in serum. Ten patients with clinical SLE and/or other collagen-vascular diseases were considered separately. Results were compared to an Arizona control group, to published series, and to laboratory controls. Frequencies of each of 10 ARA symptoms were higher in exposed subjects than in any comparison group except those with clinical SLE. The number of subjects with 4 or more symptoms was 2.3 times higher compared to referent women and men. FANA titers > 1:80 was approximately 2.3 times higher in women but equally frequent in men as in laboratory controls. ARA score and FANA rank were correlated with a coefficient (cc) of .1251, r{sup 2} = .0205 in women and this correlation was almost statistically significant in men cc = .1282, r{sup 2} = .0253. In control men and women neither correlation was significant. Long-term low-dose exposure to TCE and other chemicals in contaminated well water significantly increased symptoms of lupus erthematosus as perceived by the ARA score and the increased FANA titers.

  9. Panel reactive antibody positivity and associated HLA antibodies in Turkish renal transplant candidates.

    PubMed

    Ozdemir, Fatma Nurhan; Sezer, Siren; Akcay, Ali; Arat, Zubeyde; Turan, Munire; Gulmus, Sale; Kulah, Eyup; Haberal, Mehmet

    2004-01-01

    Pre- and post-renal transplantation panel reactive antibody (PRA) screening is associated with increased incidence of hyperacute or acute graft rejection and graft loss. This study was designed to find any relationship PRA sensitization and associated human leukocyte antigen (HLA)-specific antibodies in Turkish renal transplant candidates. We included 340 patients who were in the renal transplantation waiting list in the study. We determined PRA sensitization ratio and the associated anti-HLA IgG antibody distribution of the patient group. The PRA testing was currently performed and levels above 30% were accepted to be positive. The PRA class I positivity was determined in 24 (7%) and class II in 34 (10%) of the patients. The most frequent HLA antibodies for class I were B56, A2, A34, A1, A23, A24 and B61; and for class II were DR11, DR14, DQ7, DR10, DQ5, DR1 and DR7, respectively. From these, the increase of the numbers of anti-HLA class II antibodies was significantly correlated with the increase of PRA sensitization ratio. In conclusion, the identification of the associated HLA-specific antibodies and correlation with the Turkish population HLA antigen distribution will identify the high-risk patients who are candidates for transplantation. PMID:14967317

  10. HLA class II genes associated with anticentromere antibody in Japanese patients with systemic sclerosis (scleroderma)

    Microsoft Academic Search

    M Kuwana; Y Okano; J Kaburaki; H Inoko

    1995-01-01

    OBJECTIVE--To define further HLA class II gene associations with anticentromere antibody (ACA), a major serum antinuclear antibody in patients with systemic sclerosis (SSc). METHODS--HLA class II genes were determined using polymerase chain reaction\\/restriction fragment length polymorphisms in 94 Japanese patients with SSc (22 ACA positive and 72 ACA negative) and 50 race matched normal control subjects. RESULTS--Frequency of DQB1*0501 was

  11. Effects of cytotoxin-associated gene A (CagA) positive Helicobacter pylori infection on anti-platelet glycoprotein antibody producing B cells in patients with primary idiopathic thrombocytopenic purpura (ITP)

    PubMed Central

    Cheng, Yuan-Shan; Kuang, Li-Ping; Zhuang, Chun-Lan; Jiang, Jia-Dian; Shi, Man

    2015-01-01

    Objective: To explore the effects of cytotoxin-associated gene A (CagA) positive Helicobacter pylori (H. pylori or HP) infection on circulating B cells producing specific platelet glycoprotein antibodies and the association between therapeutic outcomes in primary idiopathic thrombocytopenic purpura (ITP) patients. Methods: A total of 76 newly diagnosed primary ITP patients were included in the study which was conducted at the first affiliated hospital of Shantou University Medical college, in Shantou city China, between January 2013 and January 2014. These patients were tested for H. pylori infection by 13C urea breath test and for anti-CagA antibody in H. pylori positive cases by enzyme-linked immunosorbent assay (ELISA) method. Anti-GPIb and anti-GPIIb/IIIa antibody-producing B cells were measured using an enzyme-linked immunospot (ELISPOT) assay in all ITP patients and 30 controls. Anti-nuclear antibody (ANA) was also detected in ITP patients. Results: The numbers of anti-GPIIb/IIIa antibody-producing B cells in HP+CagA+ patients were higher than in HP+CagA- or HP- patients. However, anti-GPIb antibody-producing B cells were found higher in HP- patients. Analysis of treatment outcomes showed that a therapeutic response was more likely in patients presenting anti-GPIIb/IIIa B cells, but the poor response was found to be associated with anti-GPIb B cells and ANA presences. Conclusion: CagA antigen of H. pylori may induce anti-GPIIb/IIIa antibodies production by a molecular mimicry mechanism. Anti-GPIIb/IIIa and anti-GPIb antibody producing B Cells detection is useful for predicting treatment effects of primary ITP. PMID:25878627

  12. Risk factors for ANA positivity in healthy persons

    Microsoft Academic Search

    Quan-Zhen Li; David R Karp; Jiexia Quan; Valerie K Branch; Jinchun Zhou; Yun Lian; Benjamin F Chong; Edward K Wakeland; Nancy J Olsen

    2011-01-01

    Introduction  The finding of antinuclear antibody (ANA) positivity in a healthy individual is usually of unknown significance and in most\\u000a cases is benign. However, a subset of such individuals is at risk for development of autoimmune disease. We examined demographic\\u000a and immunological features that are associated with ANA positivity in clinically healthy persons to develop insights into\\u000a when this marker carries

  13. Heterophilic antibodies interfering with radioimmunoassay. A false-positive pregnancy test

    SciTech Connect

    Vladutiu, A.O.; Sulewski, J.M.; Pudlak, K.A.; Stull, C.G.

    1982-11-19

    A young woman with amenorrhea had a consistently positive pregnancy test result (serum radioimmunoassay measurement of ..beta..-human chorionic gonadotropin hormone). No fetal or placental tissue was found after uterine curettage and exploratory laparotomy. The false-positive pregnancy test result was due to heterophilic antibovine and antigoat antibodies in the patient's serum. These antibodies interfered with radioimmunoassays using goat antibodies. This case shows that serum heterophilic antibodies can interfere with immunoassays and result in unnecessary diagnostic procedures and/or unnecessary treatment.

  14. Anti-aquaporin-4 antibody-positive dorsal midbrain syndrome.

    PubMed

    Lee, Juyoun; Jeong, Seong-Hae; Park, Sang Min; Sohn, Eun Hee; Lee, Ae Young; Kim, Jae-Moon; Jo, Hyun-Jin; Lee, Yeon-Hee; Kim, Ji-Soo

    2015-04-01

    Neuromyelitis optica spectrum disorders (NMOSD) can cause various ocular motor disorders in addition to optic neuritis. Ocular motor findings associated with NMOSD include spontaneous vertical and gaze-evoked nystagmus, wall-eyed bilateral internuclear ophthalmoplegia, and trochlear nerve palsy. The association between dorsal midbrain syndrome and anti-aquaporin-4 antibody seropositivity has not been reported. Here, we report a patient displaying typical dorsal midbrain syndrome and anti-aquaporin-4 antibody seropositivity. PMID:25013154

  15. Panel reactive antibody positivity and associated HLA antibodies in Turkish renal transplant candidates

    Microsoft Academic Search

    Fatma Nurhan Ozdemir; Siren Sezer; Ali Akcay; Zubeyde Arat; Munire Turan; Sale Gulmus; Eyup Kulah; Mehmet Haberal

    2004-01-01

    Pre- and post-renal transplantation panel reactive antibody (PRA) screening is associated with increased incidence of hyperacute or acute graft rejection and graft loss. This study was designed to find any relationship PRA sensitization and associated human leukocyte antigen (HLA)-specific antibodies in Turkish renal transplant candidates. We included 340 patients who were in the renal transplantation waiting list in the study.

  16. Sensitivity of the HEp2000 Substrate for the Detection of Anti-SSA\\/Ro60 Antibodies

    Microsoft Academic Search

    I. Peene; W. Van Ael; M. Vandenbossche; T. Vervaet; E. Veys; F. De Keyser

    2000-01-01

    :   Anti-SSA\\/Ro antibodies are the most prevalent type of antinuclear antibody (ANA). Anti-SSA\\/Ro-positive sera may recognise\\u000a two proteins: a 52 kDa (Ro52) and a 60 kDa (Ro60) subunit. We studied the sensitivity for Ro60 detection using the HEp-2000\\u000a substrate, which consists of HEp-2 cells transfected with Ro60 cDNA in an anti-SSA\\/Ro-positive population consecutively identified\\u000a by double immunodiffusion (DID) with thymus\\/spleen

  17. Poorly differentiated hepatocellular carcinoma accompanied by anti-Hu antibody-positive paraneoplastic peripheral neuropathy.

    PubMed

    Matsui, Takahiro; Hori, Yumiko; Nagano, Hiroaki; Eguchi, Hidetoshi; Marubashi, Shigeru; Wada, Hiroshi; Wada, Naoki; Ikeda, Jun-Ichiro; Sakamoto, Michiie; Morii, Eiichi

    2015-07-01

    The anti-Hu antibody is one of the most famous onco-neural antibodies related to paraneoplastic neurological syndrome, and is associated with small cell lung carcinoma in most cases. Here, we report a case of poorly differentiated hepatocellular carcinoma accompanied by paraneoplastic peripheral neuropathy positive for the anti-Hu antibody. Image inspection before operation revealed that no tumors were found in organs other than the liver, including lung, and that the liver tumor had no metastatic lesion. The liver tumor showed histological appearance of poorly differentiated carcinoma with cartilaginous metaplasia and partial blastoid cell appearance. Most tumor cells presented trabecular-like structure lined by sinusoidal vessels. Immunohistochemically, the tumor cells were positive for low molecular weight cytokeratin and vimentin, partially positive for cytokeratin 19 and CD56, but negative for synaptophysin, chromogranin A and alpha-fetoprotein. Based on the trabecular-like morphology and the results of immunohistochemical staining, we concluded that the tumor was diagnosed as poorly differentiated hepatocellular carcinoma. Anti-Hu antibody-positive paraneoplastic peripheral neuropathy accompanied with liver tumor is extremely rare as far as is known. The presented case indicates that poorly differentiated carcinoma has the potential to be the responsible lesion of anti-Hu antibody-positive paraneoplastic neurological syndrome and systemic work-up is important for the management of this neurological disorder. PMID:25941021

  18. Antinuclear antibodies in asthma patients- a special asthma phenotype?

    PubMed

    Agache, Ioana; Duca, Liliana; Anghel, Mariana; Pamfil, Gheorghe

    2009-03-01

    Several studies reported the appearance of asthma and autoimmune conditions in the same patient, but the clinical significance of this association was not yet assessed. One hundred asthmatic patients were observed for one year evolution with death, severe exacerbations, intake of > 1000 micrograms of beclometasone or equivalent (high ICS) and FEV1 decline >100 ml, in relation with ANA (ELISA), sputum and blood eosinophilia (EO), NSAID intolerance, BMI >25, chronic rhinosinusitis, smoking status and FEV1 <30% predicted (low FEV1). After 1 year of observation, there were 5 deaths, 28 severe asthma exacerbations requiring hospitalisations, 24 cases requiring high inhaled corticosteroid intake, and 19 patients with fast FEV1 decline (>100 ml/year). Multiple regression analysis pointed out several different independent risk factors for severe asthma evolution: for death presence of ANA (P=0.037), NSAID intolerance (P<0.001) and low FEV1 (P=0.021); for evolution with severe exacerbations ANA (p=0.011), sputum EO (P<0.001), smoking (P=0.044) and NSAID intolerance (P=0.022); for high ICS intake ANA (P=0.036), sputum EO (P=0.026) and low FEV1 (P=0.006); for FEV1 decline >100 ml ANA (P=0.006), sputum EO (P=0.037), BMI>25 (P=0.046) and NSAID intolerance (P=0.017). The presence of ANA is an independent risk factor in asthma for evolution with death, severe exacerbations, high inhaled corticosteroid intake and FEV1 decline >100 ml. PMID:19279359

  19. Hepatitis B core antibody-positive donors in cardiac transplantation: a single-center experience.

    PubMed

    Horan, J L; Stout, J E; Alexander, B D

    2014-10-01

    Hepatitis B virus (HBV) core antibody (HBcAb)-positive donors are increasingly utilized in solid organ transplantation. We report a single center's experience in cardiac transplantation with 18 HBcAb-positive donors. Available follow-up on recipients of cardiac allografts from HBcAb-positive donors, including 2 donors with low-level serum HBV DNA at the time of transplantation, demonstrated no documented donor-derived HBV transmission. PMID:25154437

  20. Studies on production of anticollagen antibodies in silicosis

    SciTech Connect

    Nagaoka, Tadasu; Tabata, Masaji; Kobayashi, Kenichi; Okada, Akira (Kanazawa Univ. (Japan))

    1993-01-01

    Silicosis is characterized by pulmonary fibrotic changes which consist primarily of an increase in collagen. In this study, anticollagen antibodies in the serum of 134 silicosis patients versus 40 normal subjects were examined and their relationship with immunoglobulin, autoantibodies, and procollagen III peptide (PIIIP) was investigated by enzyme-linked immunosorbent assay (ELISA). The mean levels of antihuman type I collagen (HI) and anti-human type Ill collagen (HIII) antibodies were significantly higher in the silicosis patients versus the normal subjects (P < 0.001). However, no differences were observed in the mean levels of anti-human type IV collagen (HIV) antibodies in the silicosis patients versus the normal subjects. Anticollagen antibodies in the sera of silicosis patients appear to be formed at an early stage of the disease. We observed a correlation between anticollagen antibodies and immunoglobulin. There was a tendency toward high values of anticollagen antibodies in the sera of patients positive for antinuclear antibodies (ANA) and rheumatoid factor (RF), both of which are autoantibodies. However, no correlation was observed between serum PIIIP and anticollagen antibodies. These observations suggest that, in silicosis, there is a relationship between anticollagen antibodies and immunoglobulins, as well as between anticollagen antibodies and autoantibodies. Measurement of anticollagen antibodies in the sera of silicosis patients offers a useful index for evaluating the prognosis of pulmonary fibrosis and autoimmune abnormality in silicosis. 49 refs. 6 figs., 6 tabs.

  1. Antineutrophilic Cytoplasmic Antibody Positive Vasculitis Associated with Methimazole Use

    PubMed Central

    Shikha, Deep; Harris, Jonathan; Resta, Christine; Park, Patricia

    2015-01-01

    ANCA-associated vasculitis (AAV) is a rare and potentially life threatening complication associated with antithyroid drug use. It is more commonly reported with propylthiouracil, with fewer cases reported with methimazole use. We present the case of a 55-year-old man with toxic multinodular goiter which was treated with methimazole for 6 months. He developed ANCA positive leukocytoclastic vasculitis with hemorrhagic and necrotic bullous lesions of lower extremities. The vasculitis was initially thought to be secondary to recent cephalosporin use; however, the skin lesions progressed despite stopping the cephalosporin and treatment with steroids, and he developed osteomyelitis. His vasculitis resolved after cessation of methimazole use. This case highlights the importance of careful monitoring for variable manifestations of AAV in patients treated with methimazole. PMID:26060588

  2. [Ataxia with ophthalmoplegia: Miller-Fisher syndrome with anti-GQ1b antibody positivity].

    PubMed

    Guilloton, L; Camarasa, C; Agard, E; Tondeur, G; Dot, C; Drouet, A

    2014-02-01

    Miller-Fisher syndrome is defined as ophthalmoplegia, ataxia and areflexia. Considered as a variant of Guillain-Barré syndrome, it differs in its clinical presentation and by anti-GQ1b antibody positivity. The authors report a case of Miller-Fisher syndrome characterized by ataxia and complete ophthalmoplegia. Through this example, the range of ophthalmologic clinical manifestations are discussed. PMID:24513384

  3. Clinical, HLA, and Small Bowel Immunohistochemical Features of Children with Positive Serum Antiendomysium Antibodies and Architecturally Normal Small Intestinal Mucosa

    Microsoft Academic Search

    Francesco Paparo; Emma Petrone; Antonella Tosco; Maria Maglio; Melissa Borrelli; Virginia M. Salvati; Erasmo Miele; Luigi Greco; Salvatore Auricchio; Riccardo Troncone

    2005-01-01

    BACKGROUND:Antiendomysium antibodies have a high sensitivity and specificity for celiac disease. A small percentage of subjects positive for these antibodies have a small intestinal mucosa hitherto considered normal.OBJECTIVES:The aim of this study was to characterize the clinical, serological, immunogenetic, and immunohistological features of these subjects.METHODS:From 409 patients who were positive for celiac-related antibodies, we selected 24 (5.9%) patients who had

  4. Isolation of Highly Active Monoclonal Antibodies against Multiresistant Gram-Positive Bacteria

    PubMed Central

    Rossmann, Friederike S.; Laverde, Diana; Kropec, Andrea; Romero-Saavedra, Felipe; Meyer-Buehn, Melanie; Huebner, Johannes

    2015-01-01

    Multiresistant nosocomial pathogens often cause life-threatening infections that are sometimes untreatable with currently available antibiotics. Staphylococci and enterococci are the predominant Gram-positive species associated with hospital-acquired infections. These infections often lead to extended hospital stay and excess mortality. In this study, a panel of fully human monoclonal antibodies was isolated from a healthy individual by selection of B-cells producing antibodies with high opsonic killing against E. faecalis 12030. Variable domains (VH and VL) of these immunoglobulin genes were amplified by PCR and cloned into an eukaryotic expression vector containing the constant domains of a human IgG1 molecule and the human lambda constant domain. These constructs were transfected into CHO cells and culture supernatants were collected and tested by opsonophagocytic assay against E. faecalis and S. aureus strains (including MRSA). At concentrations of 600 pg/ml, opsonic killing was between 40% and 70% against all strains tested. Monoclonal antibodies were also evaluated in a mouse sepsis model (using S. aureus LAC and E. faecium), a mouse peritonitis model (using S. aureus Newman and LAC) and a rat endocarditis model (using E. faecalis 12030) and were shown to provide protection in all models at a concentration of 4 ?g/kg per animal. Here we present a method to produce fully human IgG1 monoclonal antibodies that are opsonic in vitro and protective in vivo against several multiresistant Gram-positive bacteria. The monoclonal antibodies presented in this study are significantly more effective compared to another monoclonal antibody currently in clinical trials. PMID:25706415

  5. Primary Thrombosis Prophylaxis in Antiphospholipid Antibody–Positive Patients: Where Do We Stand?

    Microsoft Academic Search

    Medha Barbhaiya; Doruk Erkan

    2011-01-01

    Persistently positive antiphospholipid antibodies (aPLs) with thrombosis and\\/or pregnancy morbidity are the hallmark of the\\u000a antiphospholipid syndrome. However, aPL-positive patients with no prior history of thrombosis exist. On the basis of a limited\\u000a number of studies that predominantly included systemic lupus erythematosus patients, aPL-positive patients without previous\\u000a thrombosis have a 0% to 3.8% annual incident thrombosis risk. Given that every

  6. Positive hepatitis B virus core antibody in HIV infection--false positive or evidence of previous infection?

    PubMed

    Pallawela, S N S; Sonnex, C; Mabayoje, D; Bloch, E; Chaytor, S; Johnson, M A; Carne, C; Webster, D P

    2015-02-01

    Isolated HBV core antibody (anti-HBc) is defined as the presence of anti-HBc with a negative HBV surface antigen (HBsAg) and HBV surface antibody (anti-HBs <10?IU/l). In patients infected with HIV with isolated anti-HBc, the aim was to determine: The prevalence of isolated positive anti-HBc; The most effective method of identifying which patients have had previous Hepatitis B Virus (HBV) infection; The prevalence of false positive anti-HBc. HBV serology results were identified from 539 patients infected with HIV sampled between January 2010 and December 2012. In those with an isolated anti-HBc and negative anti-HBe, a second anti-HBc test was carried out using a different assay. Samples were also screened for HBV DNA. The anti-retroviral regimens at time of screening were documented. 101/539 had an isolated anti-HBc. Of these, 32 (32%) had a positive anti-HBe (including 1 equivocal) and 69(68%) were anti-HBe negative. Of those negative for anti-HBe, 32 were tested for both DNA and a second anti-HBc. Of these 26 (81%) were on cART at time of HBV testing, with 25 (78%) on ART with anti-HBV activity. The prevalence of isolated anti-HBc was 19%. Only 32% were also anti-HBe positive, whereas 97% of those anti-HBe negative were positive on a second anti-HBc assay suggesting lack of utility of anti-HBe in resolving serological quandaries. One subject (3%) had a false positive anti-HBc. There was no evidence of chronic HBV but 78% patients were on HBV-suppressive combination anti-retroviral therapy. PMID:25174739

  7. Investigation of False Positive Results with an Oral Fluid Rapid HIV1\\/2 Antibody Test

    Microsoft Academic Search

    Krishna Jafa; Pragna Patel; Duncan A. MacKellar; Patrick S. Sullivan; Kevin P. Delaney; Tracy L. Sides; Alexandra P. Newman; Sindy M. Paul; Evan M. Cadoff; Eugene G. Martin; Patrick A. Keenan; Bernard M. Branson; Douglas Nixon

    2007-01-01

    BackgroundIn March 2004, the OraQuick® rapid HIV antibody test became the first rapid HIV test approved by the US Food and Drug Administration for use on oral fluid specimens. Test results are available in 20 minutes, and the oral fluid test is non-invasive. From August 2004–June 2005, we investigated a sudden increase in false-positive results occurring in a performance study

  8. An effective immunotherapy regimen for VGKC antibody-positive limbic encephalitis

    Microsoft Academic Search

    S. H. Wong; M. D. Saunders; A. J. Larner; K. Das; I. K. Hart

    2010-01-01

    BackgroundVoltage-gated potassium channel antibody-positive limbic encephalitis (VGKC+LE) frequently improves with immunotherapy, although the optimum regimen is unknown. The effectiveness of a combination immunomodulatory regimen was tested in consecutive VGKC+LE patients.MethodsThis was an open-label prospective study of nine VGKC+LE patients. All patients had plasma exchange (50 ml\\/kg), intravenous immunoglobulin (2 g\\/kg) and intravenous methylprednisolone (1 g×3), followed by maintenance oral prednisolone

  9. Relationship between Antibody-Positive Rate against Plasmodium vivax Circumsporozoite Protein and Incidence of Malaria.

    PubMed

    Lee, Hyeong-Woo; Kang, Yoon-Joong; Cho, Shin-Hyeong; Na, Byoung-Kuk; Pak, Jhang Ho; Nam, Ho-Woo; Park, Yun-Kyu; Sohn, Youngjoo; Kim, Tong-Soo

    2015-04-01

    The relationship between anti- Plasmodium vivax circumsporozoite protein (CSP) antibody levels and the prevalence of malaria in epidemic areas of South Korea was evaluated. Blood samples were collected from inhabitants of Gimpo-si (city), Paju-si, and Yeoncheon-gun (county) in Gyeonggi-do (province), as well as Cheorwon-gun in Gangwon-do from November to December 2004. Microscopic examinations were used to identify malaria parasites. ELISA was used to quantitate anti-circumsporozoite protein (CSP) antibodies against P. vivax. A total of 1,774 blood samples were collected. The overall CSP-ELISA-positive rate was 7.7% (n=139). The annual parasite incidences (APIs) in these areas gradually decreased from 2004 to 2005 (1.09 and 0.80, respectively). The positive rate in Gimpo (10.4%, 44/425) was the highest identified by CSP-ELISA. The highest API was found in Yeoncheon, followed by Cheorwon, Paju, and Gimpo in both years. The positive rates of CSP-ELISA were closely related to the APIs in the study areas. These results suggest that seroepidemiological studies based on CSP may be helpful in estimating the malaria prevalence in certain areas. In addition, this assay can be used to establish and evaluate malaria control and eradication programs in affected areas. PMID:25925175

  10. Successful treatment with noninvasive positive-pressure ventilation based on the prediction of disease onset using CT and respiratory function tests in an elderly patient with relapsing polychondritis.

    PubMed

    Yamaguchi, Hiromichi; Komase, Yuko; Ono, Ayami; Morita, Akane; Ishida, Akira

    2013-01-01

    An 83-year-old man who had been receiving treatment for bronchial asthma since 62 years of age experienced difficulty breathing on exertion and was admitted to the hospital. On admission, computed tomography revealed tracheal wall thickening, while test results for antinuclear antibodies and anti-type II collagen antibodies were positive. Since a saddle nose deformity, malacia of the auricles and sensorineural deafness were also observed, relapsing polychondritis was diagnosed. Measuring the peak expiratory flow rate was useful in the early airway assessment. During the follow-up period, the patient's dyspnea worsened and noninvasive positive-pressure ventilation was introduced. As a result, the subjective symptoms improved. PMID:23676595

  11. Comparison of Antibodies That Mediate HIV Type 1 gp120 Antibody-Dependent Cell-Mediated Cytotoxicity in Asymptomatic HIV Type 1-Positive Men and Women

    PubMed Central

    Mata, Mariana M.; Iwema, Joyce R.; Dell, Shanna; Neems, Leslie; Jamieson, Beth D.; Phair, John; Cohen, Mardge H.; Anastos, Kathryn

    2014-01-01

    Abstract Recent studies suggest that HIV-specific antibody-dependent cell-mediated cytotoxicity (ADCC) antibodies contribute to protective immunity against HIV. An important characteristic of future HIV vaccines will, therefore, be the ability to stimulate production of these antibodies in both men and women. Early studies suggest that men may have a better ADCC antibody response against HIV than women. Our objective was to determine whether men and women differ with respect to their ADCC response to HIV-1 gp120. HIV-positive, asymptomatic untreated men and women were matched for race, age, CD4+ T cell number, HIV-1 viral load, and treatment and HIV-1 gp120 ADCC antibody titers were compared. A standard 51Cr-release assay was used to determine HIV-1 gp120 ADCC antibody titers in HIV-1-seropositive individuals from the Multicenter AIDS Cohort Study (MACS; n=32) and the Women's Interagency HIV Study (WIHS; n=32). Both sexes had high ADCC titers against HIV-1 gp120: 34.4% (n=11) and 40.6% (n=13) of men and women, respectively, had titers of 10,000; 62.5% (n=20) and 56.3% (n=18) had titers of 100,000. Groups did not differ in percent specific release (% SR), lytic units (LU), correlations of titer to viral load, or titer to CD4+ T cells in men or women. Both groups also had similar cross-clade ADCC antibody responses (p>0.5 for % SR and LU). Comparable groups of asymptomatic HIV-1-infected men and women had comparable HIV-1 gp120 ADCC antibodies. Both sexes had significant cross-clade reactivity. Differences between men and women may become evident as disease progresses; this should be evaluated at later stages of HIV-1 infection. PMID:23972002

  12. Monoclonal Antibody Against the Turn of the 42-Residue Amyloid ?-Protein at Positions 22 and 23

    PubMed Central

    2010-01-01

    Aggregation of the 42-mer amyloid ?-protein (A?42) plays a critical role in the pathogenesis of Alzheimer’s disease (AD). We have proposed a toxic conformer with a turn at positions 22 and 23, as well as a nontoxic conformer with a turn at positions 25 and 26, in A?42 aggregates from systematic proline scanning and solid-state NMR studies. Although recent clinical trials of immunization targeting A?42 aggregates have proved useful, some adverse effects were reported. One of the reasons was hypothesized to be excessive immunoreactions derived from the unintended removal of nontoxic A?42, which plays an important role in the physiological function. To develop a monoclonal antibody for toxic A?42, E22P-A?10-35, a minimum moiety for neurotoxicity containing the turn at positions 22 and 23, was used for the generation of antibodies, following the selection of clones using A?42 mutants of E22P (turn-inducing) and E22V (turn-preventing). The obtained clone (11A1) showed a high binding affinity (KD = 10.3 nM) for A?42 using surface plasmon resonance. 11A1 also inhibited the neurotoxicity of A?42 in PC12 cells. Immunohistochemical studies showed that not only extracellular but intracellular amyloid was stained in human AD brains. In Western blotting analyses using human brains, low-molecular weight-oligomers rather than the monomer of A? were readily recognized by 11A1. These results imply that 11A1 could detect toxic A?42 oligomers with the turn at positions 22 and 23 and that 11A1 could be applicable for the therapeutic targeting of toxic A?42 in AD. PMID:22778811

  13. Antineutrophil cytoplasmic antibody-positive conversion and microscopic polyangiitis development in patients with idiopathic pulmonary fibrosis

    PubMed Central

    Kagiyama, Naho; Takayanagi, Noboru; Kanauchi, Tetsu; Ishiguro, Takashi; Yanagisawa, Tsutomu; Sugita, Yutaka

    2015-01-01

    Background Increasing evidence indicates that antineutrophil cytoplasmic antibody (ANCA)-positive conversion occurs in patients initially diagnosed with idiopathic pulmonary fibrosis (IPF) and as a result, some of these patients develop microscopic polyangiitis (MPA). However, the incidence density of these patients is not well known. Objectives To explore the incidence of ANCA-positive conversion and development of MPA during the disease course in patients with IPF and to evaluate whether corticosteroid therapy reduces MPA development in patients with IPF with myeloperoxidase (MPO)-ANCA positivity at diagnosis or who later acquire MPO-ANCA positivity. Methods We retrospectively analysed the medical records of 504 Asian patients with IPF treated at our institution in Saitama, Japan. Results Of the 504 patients with IPF, 20 (4.0%) had MPO-ANCA and 16 (3.2%) had PR-3-ANCA when first evaluated. In 264 of 504 patients with IPF, ANCA was measured repeatedly and seroconversion to MPO-ANCA and PR3-ANCA occurred in 15 (5.7%) and 14 (5.3%) patients, respectively, and 9 of 35 patients who were either MPO-ANCA positive at IPF diagnosis or who subsequently seroconverted developed MPA. None of the nine patients who developed MPA had been previously treated with steroids. The incidence of MPA tended to be lower in patients treated than not treated with corticosteroids although this was not statistically significant. Conclusions Some patients with IPF with MPO-ANCA positivity at IPF diagnosis or with MPO-ANCA-positive conversion during follow-up developed MPA. Clinical trials to determine whether corticosteroid therapy can reduce MPA development and prolong survival in MPO-ANCA-positive patients with IPF should be considered. PMID:25593704

  14. Antithyroid peroxidase antibody positivity is associated with lower incidence of metastasis in breast cancer

    PubMed Central

    KEMAL, YASEMIN; DEMIRAG, GUZIN; EKIZ, KUBILAY; YUCEL, IDRIS

    2015-01-01

    Thyroid extracts were first used to treat patients with metastatic breast cancer over a century ago. Since then, a number of studies have investigated the association between thyroid disorders and breast cancer. The presence of antibodies to thyroid peroxidase (TPOab) was recently reported to be associated with improved outcome in these patients. The aim of the present study was to evaluate the association between TPOab positivity and clinicopathological characteristics in breast cancer patients. The study included 318 newly diagnosed cases of breast cancer treated at Ondokuz Mayis University Hospital, Samsun, Turkey, between 2008 and 2012. Serum thyroid-stimulating hormone, free triiodothyronine and free thyroxine levels were measured at the time of diagnosis. Of the 318 patients, 253 were considered to be TPOab-negative (TPOab ?34 IU/ml) and 65 TPOab-positive (TPOab >34 IU/ml). No cases with distant metastases were found in the TPOab-positive group. However, 20 (7.9%) of the 253 patients displayed distant metastases in the TPOab-negative group (P=0.01). Therefore, TPOab positivity was found to be associated with a lower incidence of metastasis in breast cancer patients.

  15. Investigation of False Positive Results with an Oral Fluid Rapid HIV-1/2 Antibody Test

    PubMed Central

    Jafa, Krishna; Patel, Pragna; MacKellar, Duncan A.; Sullivan, Patrick S.; Delaney, Kevin P.; Sides, Tracy L.; Newman, Alexandra P.; Paul, Sindy M.; Cadoff, Evan M.; Martin, Eugene G.; Keenan, Patrick A.; Branson, Bernard M.

    2007-01-01

    Background In March 2004, the OraQuick® rapid HIV antibody test became the first rapid HIV test approved by the US Food and Drug Administration for use on oral fluid specimens. Test results are available in 20 minutes, and the oral fluid test is non-invasive. From August 2004–June 2005, we investigated a sudden increase in false-positive results occurring in a performance study of OraQuick® oral-fluid rapid HIV tests in Minnesota. Methodology/Principal Findings In a field investigation, we reviewed performance study data on oral-fluid and whole-blood OraQuick® rapid HIV test device lots and expiration dates and assessed test performance and interpretation with oral-fluid and whole-blood specimens by operators who reported false-positive results. We used multivariate logistic regression to evaluate client demographic and risk characteristics associated with false-positive results. Next, we conducted an incidence study of false-positive OraQuick rapid HIV tests in nine US cities and tested both oral-fluid and finger-stick whole-blood specimens from clients; reactive tests were confirmed with Western blot. Sixteen (4.1%) false-positive oral-fluid results occurred in the performance study from April 15, 2004 through August 31, 2004 with unexpired devices from six test lots among 388 HIV-uninfected clients (specificity, 95.9%; 95% CI: 93.4–97.6). Three test operators who had reported false-positive results performed and interpreted the test according to package-insert instructions. In multivariate analysis, only older age was significantly associated with false-positive results (adjusted odds ratio?=?4.5, 95% CI: 1.2–25.7). In the incidence study, all valid oral-fluid and whole-blood results from 2,268 clients were concordant and no false-positive results occurred (100% specificity). Conclusions/Significance The field investigation did not identify a cause for the increase in false-positive oral-fluid results, and the incidence study detected no false-positive results. The findings suggest this was an isolated cluster; the test's overall performance was as specified by the manufacturer. PMID:17268576

  16. Acute Liver Failure in an Antimitochondrial Antibody-Positive 63-Year-Old Man

    PubMed Central

    Wakamatsu, Toru; Kanda, Tatsuo; Tawada, Akinobu; Miyamura, Tatsuo; Takahashi, Masanori; Chiba, Tetsuhiro; Arai, Makoto; Maruyama, Hitoshi; Fujiwara, Keiichi; Imazeki, Fumio; Yokosuka, Osamu

    2012-01-01

    Antimitochondrial antibody (AMA) is one of the representative features of primary biliary cirrhosis (PBC). PBC is a female-dominant disease usually presenting intrahepatic bile duct destruction, cholestasis and fibrosis with or without chronic nonsuppurative destructive cholangitis. We presented the case of a 63-year-old man with acute liver failure who had AMA, pronounced alanine aminotransferase elevation and high bilirubinemia. We administered corticosteroids and rescued this patient without liver transplantation. It is well known that some patients within the spectrum of autoimmune liver disease present with characteristics of both PBC and autoimmune hepatitis. Although corticosteroids may be associated with a significant worsening of adverse events in patients with PBC, if acute liver failure in AMA-positive cases is progressive, the administration of corticosteroids has to be considered, as well as the preparation of urgent liver transplantation. PMID:22933985

  17. Remarkably increased resistin levels in anti-AChR antibody-positive myasthenia gravis.

    PubMed

    Zhang, Da-Qi; Wang, Rong; Li, Ting; Li, Xin; Qi, Yuan; Wang, Jing; Yang, Li

    2015-06-15

    Resistin is a pro-inflammatory cytokine involved in the pathogenesis of autoimmune diseases. To investigate serum resistin levels in patients with myasthenia gravis (MG) and determine if there are associations between resistin levels and disease severity, we measured serum resistin levels in 102 patients with anti-acetylcholine receptor antibody-positive MG (AChR-MG). We further analyzed associations between serum resistin levels and clinical variables in patients with MG. Our findings demonstrate that serum resistin levels are elevated in patients with AChR-generalized MG and AChR-MG with thymoma and are correlated with disease severity. Resistin has potential as a useful serum biomarker for inflammation in AChR-MG. PMID:26004149

  18. [Serum IgG antibodies to Chlamydia trachomatis in HLA-B-27 positive patients with rheumatic diseases].

    PubMed

    Breustedt, W; Giesel, A; Puhlmann, B

    1989-01-01

    Urogenital chlamydial infection should act as a trigger of peripheral and axial form of arthritis like sexually acquired reactive arthritis and Reiter's syndrome assumedly. HLA B-27 positive patients with rheumatic disorders, mainly M. Bechterew (55 out of 62 men and 18 out of 25 women) were investigated for serum-IgG-antibodies to Chlamydia trachomatis (ELISA). Among HLA B-27 positive men and women urogenital disorders in their history were more common as compared with controls. There were found no differences concerning the rate of antichlamydial antibodies between patients and controls. It is supposed, that urogenital bacterial infections rather others than chlamydial infection may be connected with M. Bechterew. PMID:2792489

  19. Central retinal vein occlusion in a pediatric patient with SLE and antiphospholipid antibodies without anti-cardiolipin or anti-?2 glycoprotein I antibodies

    PubMed Central

    2014-01-01

    Background Antiphospholipid antibody syndrome is characterized by venous and/or arterial thrombosis, and is found in patients with systemic lupus erythematosus. Its diagnosis requires the presence of both clinical and laboratory findings, such as positive anti-cardiolipin and anti-?2 glycoprotein I antibodies and lupus anticoagulant. However, cardiolipin is a minor component of the vascular endothelial cells in human, and phosphatidylcholine and phosphatidylethanolamine are major components. Case presentation A 15-year-old female suddenly developed massive left intraretinal hemorrhaging due to central retinal vein occlusion. She also had a butterfly rash, and her laboratory findings revealed positive serum anti-nuclear antibodies and decreased serum complement. During this episode, she was diagnosed with systemic lupus erythematosus. Although she was negative for serum anti-cardiolipin IgG and anti-?2 glycoprotein I antibodies as well as lupus anticoagulant, her serum anti-phosphatidylcholine, anti-phosphatidylethanolamine, anti-phosphatidylinositol and phosphatidylserine IgG antibodies levels were increased. Conclusion Pediatric cases of central retinal vein occlusion are rare. Even in patients without anti-cardiolipin or anti-?2 glycoprotein I antibodies and lupus anticoagulant, there is the potential for the development of antiphospholipid antibody-related thrombosis. PMID:24885875

  20. The Drug User's Identity and How It Relates to Being Hepatitis C Antibody Positive: A Qualitative Study

    ERIC Educational Resources Information Center

    Copeland, Lorraine

    2004-01-01

    The increasing health problem of hepatitis C virus infection has only recently attracted the attention of psychosocial research, especially among subjects at higher risk (e.g. injecting drug users). There is a lack of information about the knowledge, perceptions and feelings that injecting drug users hold about their hepatitis C antibody positive

  1. Evaluation of Disulfide Bond Position to Enhance the Thermal Stability of a Highly Stable Single Domain Antibody

    PubMed Central

    Zabetakis, Dan; Olson, Mark A.; Anderson, George P.; Legler, Patricia M.; Goldman, Ellen R.

    2014-01-01

    Single domain antibodies are the small recombinant variable domains derived from camelid heavy-chain-only antibodies. They are renowned for their stability, in large part due to their ability to refold following thermal or chemical denaturation. In addition to refolding after heat denaturation, A3, a high affinity anti-Staphylococcal Enterotoxin B single domain antibody, possesses a melting temperature of ?84°C, among the highest reported for a single domain antibody. In this work we utilized the recently described crystal structure of A3 to select locations for the insertion of a second disulfide bond and evaluated the impact that the addition of this second bond had on the melting temperature. Four double-disulfide versions of A3 were constructed and each was found to improve the melting temperature relative to the native structure without reducing affinity. Placement of the disulfide bond at a previously published position between framework regions 2 and 3 yielded the largest improvement (>6°C), suggesting this location is optimal, and seemingly provides a universal route to raise the melting temperature of single domain antibodies. This study further demonstrates that even single domain antibodies with extremely high melting points can be further stabilized by addition of disulfide bonds. PMID:25526640

  2. Guillain-Barré syndrome-like-onset neurosarcoidosis positive for immunoglobulin G anti-N-acetylgalactosaminyl-GD1a antibody.

    PubMed

    Chatani, H; Tanaka, M; Nagata, T; Araki, T; Kusunoki, S

    2014-01-01

    Anti-ganglioside antibodies have been reported in various peripheral neuropathies, including Guillain-Barré syndrome (GBS), chronic inflammatory demyelinating polyneuropathy, multifocal motor neuropathy, Fisher syndrome, monoclonal gammopathy-associated neuropathy, and other idiopathic neuropathies. To our knowledge, there has been no report of anti-ganglioside-positive sarcoidosis. We report a 62-year-old man with acute weakness of the limbs and sensory disturbance of the right arm and trunk resembling GBS. Soluble interleukin-2 receptor and angiotensin-converting enzyme levels were elevated. Anti-ganglioside antibodies (immunoglobulin G anti-N-acetylgalactosaminyl-GD1a antibody [IgG anti-GalNAc-GD1a antibody]) were detected. Neurophysiological examination demonstrated axonal neuropathy. Bilateral hilar lymphadenopathy was demonstrated on a chest CT scan, and abnormal uptake of 67 Gallium was detected by scintigraphy. The ratio of CD4 to CD8 was elevated in bronchoalveolar lavage fluid. Noncaseating epithelioid cell granulomas were detected in a specimen obtained via transbronchial lung biopsy. Because intravenous immunoglobulin did not improve the symptoms, we commenced steroid pulse therapy followed by oral prednisolone therapy. After steroid therapy, he recovered fully. Because the findings in our patient fulfilled the criteria for neurosarcoidosis, we diagnosed his illness as probable neurosarcoidosis. To the best of our knowledge, this is the first patient with GBS-like-onset neurosarcoidosis positive for anti-IgG anti-GalNAc-GD1a antibody. PMID:23916762

  3. Biopsy findings on a stable recipient after secondary donor specific antibody (DSA) positive and ABO incompatible kidney transplantation.

    PubMed

    Sugitani, Atsushi; Takahashi, Chihiro; Naka, Takuji; Hisamitsu, Kazunori; Yamamoto, Osamu; Kobayashi, Naoto; Kimura, Mari; Yoshida, Haruhiko; Hanaki, Takehiko; Hamazoe, Ryuichi

    2015-07-01

    Using desensitization protocol, we performed a secondary donor specific antibody (DSA) positive and ABO incompatible kidney transplantation. One-hour biopsy showed no C4d deposition. The protocol biopsy after 2 weeks showed diffuse C4d deposition with peritubulitis. After 12 weeks, however, the protocol biopsy showed disappearance of tubulitis in spite of remaining C4d deposition. The recipient was in stable condition with excellent graft function despite high titer of the DSA. Monitoring of protocol biopsy is critical while antibody titer and the interpretation of the histological findings correlating with clinical markers must be considered. PMID:26031593

  4. Intragraft gene expression in positive crossmatch kidney allografts: ongoing inflammation mediates chronic antibody-mediated injury.

    PubMed

    Dean, P G; Park, W D; Cornell, L D; Gloor, J M; Stegall, M D

    2012-06-01

    We studied intragraft gene expression profiles of positive crossmatch (+XM) kidney transplant recipients who develop transplant glomerulopathy (TG) and those who do not. Whole genome microarray analysis and quantitative rt-PCR were performed on RNA from protocol renal allograft biopsies in three groups: (1) +XM/TG+ biopsies before and after TG; (2) +XM/NoTG; and (3) negative crossmatch kidney transplants (control). Microarray comparisons showed few differentially expressed genes between paired biopsies from +XM/TG+ recipients before and after the diagnosis of TG. Comparing +XM/TG+ and control groups, significantly altered expression was seen for 2447 genes (18%) and 3200 genes (24%) at early and late time points, respectively. Canonical pathway analyses of differentially expressed genes showed inflammatory genes associated with innate and adaptive immune responses. Comparing +XM/TG+ and +XM/NoTG groups, 3718 probe sets were differentially expressed but these were over-represented in only four pathways. A classic accommodation phenotype was not identified. Using rt-PCR, the expression of inflammatory genes was significantly increased in +XM/TG+ recipients compared to the +XM/NoTG and control groups. In conclusion, pretransplant donor-specific anti-HLA antibodies results in a gene expression profile characterized by inflammation and cellular infiltration and the majority of +XM grafts are exposed to chronic injury. PMID:22335458

  5. Clinical relevance of positive voltage-gated potassium channel (VGKC)-complex antibodies: experience from a tertiary referral centre

    PubMed Central

    Paterson, Ross W; Zandi, Michael S; Armstrong, Richard; Vincent, Angela; Schott, Jonathan M

    2014-01-01

    Background Voltage-gated potassium channel (VGKC)-complex antibodies can be associated with a range of immunotherapy-responsive clinical presentations including limbic encephalitis, Morvan's syndrome and acquired neuromyotonia. However, there are patients with positive levels in whom the significance is uncertain. Objective To evaluate the clinical significance associated with positive (>100?pM) VGKC-complex antibodies. Methods Over a 4-year period, 1053 samples were sent for testing of which 55 were positive. The clinical presentations, final diagnoses and responses to immunotherapies, when given, were assessed retrospectively and the likelihood of autoimmunity was categorised as definite, possible, unlikely or undetermined (modified from Zuliani et al 2012). Results Only 4 of the 32 patients with low-positive (100–400?pM) levels were considered definitely autoimmune, 3 with peripheral nerve hyperexcitability and 1 with a thymoma; 3 were given immunotherapies. Of the remaining 28 with low-positive levels, 13 (3 of whom had tumours) were considered possibly autoimmune, and 15 were unlikely or undetermined; 1 was given immunotherapy unsuccessfully. Of the 23 patients with high-positive (>400?pM) levels, 12 were given immunotherapies, 11 of whom showed a good response. 11 were considered definitely autoimmune, 10 with limbic encephalitis (antibody specificity: 5 LGI1, 1 contactin2, 2 negative, 2 untested) and 1 with a tumour. In the remaining 12, autoimmunity was considered possible (n=9; most had not received immunotherapies), or unlikely (n=3). Conclusions As antibody testing becomes more widely available, and many samples are referred from patients with less clear-cut diagnoses, it is important to assess the utility of the results. VGKC-complex antibodies in the range of 100–400?pM (0.1–0.4?nM) were considered clinically relevant in rare conditions with peripheral nerve hyperexcitability and appeared to associate with tumours (12.5%). By contrast high-positive (>400?pM; >0.4?nM) levels were considered definitely (38%) or possibly (49%) clinically relevant, but not all patients had a ‘classical’ limbic encephalitis and some did not receive immunotherapies. PMID:23757422

  6. Biological Evaluation of 131I- and CF750-Labeled Dmab(scFv)-Fc Antibodies for Xenograft Imaging of CD25-Positive Tumors

    PubMed Central

    Fan, Qing; Cai, Huawei; Yang, Hao; Li, Lin; Yuan, Cen; Lu, Xiaofeng; Wan, Lin

    2014-01-01

    A Dmab(scFv)-Fc antibody containing the single chain variable fragment of a humanized daclizumab antibody and the Fc fragment of a human IgG1 antibody was produced via recombinant expression in Pichia pastoris. The Dmab(scFv)-Fc antibody forms a dimer in solution, and it specifically binds CD25-positive tumor cells and tumor tissues. For tumor imaging, the Dmab(scFv)-Fc antibody was labeled with the 131I isotope and CF750 fluorescent dye, respectively. After intravenous injection of mice bearing CD25-positive tumor xenografts, tumor uptake of the 131I-Dmab(scFv)-Fc antibody was visible at 1?h, and clear images were obtained at 5?h using SPECT/CT. After systemic administration of the CF750-Dmab(scFv)-Fc antibody, tumor uptake was present as early as 1?h, and tumor xenografts could be kinetically imaged within 9?h after injection. These results indicate that the Dmab(scFv)-Fc antibody rapidly and specifically targets CD25-positive tumor cells, suggesting the potential of this antibody as an imaging agent for the diagnosis of lymphomatous-type ATLL. PMID:24864244

  7. A case of immunohistochemical false positive staining caused by incompatibility between a CD4 antibody and an autostainer

    PubMed Central

    Matsuda, Ikuo; Sugihara, Nao; Yunokizaki, Hiroshi; Abe, Takashi; Hirota, Seiichi

    2015-01-01

    Precise immunophenotyping of tumor cells by immunohistochemistry is complementary to morphological examination. It is critical for the correct histopathological diagnosis of lymphomas. In this paper, we report a case of T-cell lymphoma whose histopathological diagnosis was confounded by an immunohistochemical pitfall: a false positive caused by incompatibility between an antibody and an autostainer. In this case, based on CD4 immunohistochemistry of the affected lymph nodes, the T-cell lymphoma was diagnosed as CD4-positive at the onset, while it appeared discordantly to be CD4-negative at the second relapse. We noticed that CD4 antibodies and autostainers of different suppliers (designated as suppliers X and Y) were used in an unqualified combination in immunohistochemistry at the onset: that is, the combination of an antibody supplied by X and an autostainer supplied by Y (designated as X-Y combination) was used at the onset. On the other hand, the Y-Y combination was at the second relapse. At the second relapse, flow cytometry of the affected lymph node showed infiltration of CD4-negative T-cell lymphoma. We reasoned that CD4 immunonegativity obtained by the Y-Y combination at the second relapse was specific, while CD4 immunopositivity by the X-Y combination at the onset was false positive. Immunohistochemical reexamination of the lymph node at the onset proved to be CD4-negative by not only the Y-Y but also X-X combinations, confirming our final diagnosis of nodal relapse of CD4-negative T-cell lymphoma. This case illustrates the importance of using compatible combinations of antibodies and autostainers in diagnostic immunohistochemistry. PMID:25755812

  8. Steroid-Responsive Chronic Cerebellitis With Positive Glutamate Receptor ?2 Antibody

    Microsoft Academic Search

    Masaya Kubota; Yukitoshi Takahashi

    2008-01-01

    We report the clinical course of a 4-year-old girl with chronic cerebellitis (onset 2 days after diphtheria-pertussis-tetanus vaccination at 1 year and 7 months old) associated with anti-glutamate receptor ?2 antibody, who improved dramatically with steroid therapy (methylprednisolone pulse therapy plus oral prednisolone). Recently, it has been reported that the anti-glutamate receptor ?2 selectively expressed at the post-synaptic site of

  9. Anti-Ri antibody positive opsoclonus-myoclonus in a male patient with breast carcinoma

    Microsoft Academic Search

    Paul W. Wirtz; Peter A. E. Sillevis Smitt; Jorrit I. Hoff; Bertie de Leeuw; Gert Jan Lammers; Sjoerd G. van Duinen; Jan J. Verschuuren

    2002-01-01

    .   A 65-year-old male patient developed truncal ataxia, opsoclonus and myoclonus. In the serum anti-Ri antibodies were found,\\u000a which led to the detection of a small adenocarcinoma of the breast. Other prominent clinical features were an excessive startle\\u000a response and behavioral disorders, such as anxiety and impatience. These features suggest an immune response against both\\u000a Nova-1 and Nova-2 antigens throughout

  10. HLA-DRB1 Genotypes and the Risk of Developing Anti Citrullinated Protein Antibody (ACPA) Positive Rheumatoid Arthritis

    PubMed Central

    Balandraud, Nathalie; Picard, Christophe; Reviron, Denis; Landais, Cyril; Toussirot, Eric; Lambert, Nathalie; Telle, Emmanuel; Charpin, Caroline; Wendling, Daniel; Pardoux, Etienne; Auger, Isabelle; Roudier, Jean

    2013-01-01

    Objective To provide a table indicating the risk for developing anti citrullinated protein antibody (ACPA) positive rheumatoid arthritis (RA) according to one’s HLA-DRB1 genotype. Methods We HLA-DRB1 genotyped 857 patients with ACPA positive RA and 2178 controls from South Eastern and Eastern France and calculated Odds Ratios (OR) for developing RA for 106 of 132 possible genotypes accounting for 97% of subjects. Results HLA-DRB1 genotypic ORs for developing ACPA positive RA range from 28 to 0.19. HLA-DRB1 genotypes with HLA-DRB1*04SE (HLA-DRB1*0404, HLA-DRB1*0405, HLA-DRB1*0408), HLA-DRB1*04?01, HLA-DRB1*01 are usually associated with high risk for developing RA. The second HLA-DRB1 allele in genotype somewhat modulates shared epitope associated risk. We did not identify any absolutely protective allele. Neither the Reviron, nor the du Montcel models accurately explains our data which are compatible with the shared epitope hypothesis and suggest a dosage effect among shared epitope positive HLA-DRB1 alleles, double dose genotypes carrying higher ORs than single dose genotypes. Conclusion HLA-DRB1 genotypic risk for developing ACPA positive RA is influenced by both HLA-DRB1 alleles in genotype. We provide an HLA-DRB1 genotypic risk table for ACPA positive RA. PMID:23737967

  11. Detection of thyroglobulin in antithyroglobulin antibody-positive sera by isoelectric focusing.

    PubMed

    Malvoisin, Etienne; Makhloufi, Djamila; Livrozet, Jean-Michel

    2015-07-20

    Circulating antibodies have the potential to interfere with the measurement of thyroglobulin (Tg) in sera of patients. Here, we determined Tg concentration by isoelectric focusing (IEF) on agarose gel using for detection a rabbit antiserum to human Tg termed FLX. Tg was determined in sera of thyroid patients and HIV-infected patients under antiviral therapy. We showed that Tg IEF was not affected by the presence of anti-Tg antibodies (TgAb). Tg concentrations measured by IEF in TgAb-negative sera were in most of the cases, similar to those obtained by IRMA (immunoradiometric assay). However, in 5 of the 96 thyroid patients, and none of the 46 healthy subjects, Tg was undetectable by antiserum FLX and measurable by IRMA. In HIV-infected patients (64 men and 60 women), Tg was not recognized by FLX in 23 men and 9 women and this was related to abnormal CD4. We hypothesize that the decreased binding of FLX to Tg may be the result of conformational change on the Tg molecule, a phenomenon apparently related to immunodeficiency in HIV-infected patients. For thyroid patients, Tg IEF may be very useful for the interpretation of results when Tg measurements by IRMA and automated immunoassays are affected by interferences. PMID:25998693

  12. Sensitivity of the HEp-2000 substrate for the detection of anti-SSA/Ro60 antibodies.

    PubMed

    Peene, I; Van Ael, W; Vandenbossche, M; Vervaet, T; Veys, E; De Keyser, F

    2000-01-01

    Anti-SSA/Ro antibodies are the most prevalent type of antinuclear antibody (ANA). Anti-SSA/ Ro-positive sera may recognise two proteins: a 52 kDa (Ro52) and a 60 kDa (Ro60) subunit. We studied the sensitivity for Ro60 detection using the HEp-2000 substrate, which consists of HEp-2 cells transfected with Ro60 cDNA in an anti-SSA/Ro-positive population consecutively identified by double immunodiffusion (DID) with thymus/spleen nuclear extract and line immunoassay (LIA) with recombinant Ro52 and Ro60. One hundred and twenty-seven consecutive anti-SSA/Ro-positive sera defined by DID with thymus/spleen nuclear extract and LIA using recombinant Ro52 and Ro60 were analysed on HEp-2000 and DID with natural Ro60. Of these, 91 were anti-Ro60 positive on LIA and/ or DID with natural Ro60. The HEp-2000 substrate detected 70/91 (sensitivity 77%) and correlated strongest with DID. Most of the missed anti-Ro60-positive sera had high ANA intensity. The substrate did not detect monospecific anti-Ro52 antibodies (sensitivity 9.7%; 3/31). HEp-2000 substrate can therefore be considered a reliable, simple and alternative method for DID in the detection of anti-Ro60 reactivity. Special follow-up should be given to sera with strong ANA patterns in which the SSA/Ro60 staining pattern may be hidden. PMID:10941811

  13. A functional variant of Fc? receptor IIIA is associated with rheumatoid arthritis in individuals who are positive for anti-glucose-6-phosphate isomerase antibodies

    PubMed Central

    Matsumoto, Isao; Zhang, Hua; Muraki, Yoshifumi; Hayashi, Taichi; Yasukochi, Takanori; Kori, Yuko; Goto, Daisuke; Ito, Satoshi; Tsutsumi, Akito; Sumida, Takayuki

    2005-01-01

    Anti-glucose-6-phosphate isomerase (GPI) antibodies are known to be arthritogenic autoantibodies in K/B×N mice, although some groups have reported that few healthy humans retain these antibodies. The expression of Fc? receptors (Fc?Rs) is genetically regulated and has strong implications for the development of experimental arthritis. The interaction between immune complexes and Fc?Rs might therefore be involved in the pathogenesis of some arthritic conditions. To explore the relationship between functional polymorphisms in Fc?Rs (FCGR3A-158V/F and FCGR2A-131H/R) and arthritis in individuals positive for anti-GPI antibodies, we evaluated these individuals with respect to FCGR genotype. Genotyping for FCGR3A-158V/F and FCGR2A-131H/R was performed by PCR amplification of the polymorphic site, followed by site specific restriction digestion using the genome of 187 Japanese patients with rheumatoid arthritis (including 23 who were anti-GPI antibody positive) and 158 Japanese healthy individuals (including nine who were anti-GPI antibody positive). We report here on the association of FCGR3A-158V/F functional polymorphism with anti-GPI antibody positive status. Eight out of nine healthy individuals who were positive for anti-GPI antibodies possessed the homozygous, low affinity genotype FCGR3A-158F (odds ratio = 0.09, 95% confidence interval 0.01–0.89; P = 0.0199), and probably were 'protected' from arthritogenic antibodies. Moreover, among those who were homozygous for the high affinity genotype FCGR3A-158V/V, there were clear differences in anti-human and anti-rabbit GPI titres between patients with rheumatoid arthritis and healthy subjects (P = 0.0027 and P = 0.0015, respectively). Our findings provide a molecular model of the genetic regulation of autoantibody-induced arthritis by allele-specific affinity of the Fc?Rs. PMID:16277670

  14. Available means: manifestations of Aristotle's three modes of rhetorical appeal in antinuclear fiction

    SciTech Connect

    Mannix, P.J.

    1986-01-01

    The abundance of sympathetic scientists, military men and clergymen in antinuclear fiction reflects a public perception that authorities speak most knowledgeably about an issue. Other antinuclear works employ characters with less traditional ethical appeals: nurturing women, vital youths, and even infallible computers. Antinuclear fiction uses enthymeme and example to reflect the history of the nuclear weapons debate. Some works attach the immorality of the weapons by examining the moral dilemmas of nuclear scientists. Others admit the permanence of the nuclear threat. By arousing emotions, fiction is capable of mobilizing its audience's active support for the ideas it presents. The principal emotions that various antinuclear works arouse highlight the close relationship between literature and rhetoric. The most dominant emotions, pity and fear, are the two Aristotle links to tragedy. Scorn, the principal emotion that Dr. Strangelove arouses - is the crucial emotion on which all satire depends. However, the other principal emotion in anti-nuclear fiction - hope - has principally a rhetorical function ensuring that the feelings the works provoke will be channeled constructively.

  15. Prevalence of antibodies against Treponema pallidum among HIV-positive patients in a tertiary care hospital in Mexico.

    PubMed

    Mata-Marín, José Antonio; Sandoval-Sánchez, Juan Joel; Huerta-García, Gloria; Arroyo-Anduiza, Carla Ileana; Alcalá-Martínez, Enrique; Mata-Marín, Luis Alberto; Sandoval-Ramirez, Jorge Luis; Gaytán-Martínez, Jesús

    2015-02-01

    Our objective was to determine the seroprevalence of syphilis among HIV-infected patients in a tertiary care hospital in Mexico City. A cross-sectional study was developed, and 318 HIV-positive patients were evaluated from January to February 2013 at Hospital de Infectología, National Medical Center 'La Raza' (a tertiary care hospital specialising in infectious diseases in Mexico City). Laboratory data were screened for the detection of antibodies against Treponema pallidum. Patients completed a questionnaire relating to socio-demographic data and factors associated with syphilis. Of the 318 patients, 83% were men. The mean age ± SD was 36?±?11 years; 52% were men who have sex with men and 47% had undertaken higher education. The overall seroprevalence of syphilis among these patients was 25% (95% confidence interval 21%, 30%). Men who have sex with men had a significantly higher seroprevalence (30% vs. 15%, p?=?0.009). We conclude that, in Mexico, there is a high seroprevalence of syphilis antibodies in HIV-infected patients and that men who have sex with men are the group most affected. PMID:24713227

  16. Prevalence and Risk Factors for Neutralizing Antibodies to Human Papillomavirus Types 16 and 18 in HIV-positive Men who have Sex with Men

    PubMed Central

    Sharma, Rachna; Efird, Jimmy T.; Chein, Aung; Holly, Elizabeth A.; Krajden, Mel; Berry, Michael J.; Darragh, Teresa M.; Jay, Naomi; Palefsky, Joel M.

    2013-01-01

    Objective HPV vaccination is routinely recommended in HIV-positive MSM ? 26 years old. Levels of prior HPV exposure in older HIV-positive MSM are assumed to be too high to warrant routine HPV vaccination. However, little is known about the prevalence of and risk factors for neutralizing antibody seropositivity to HPV-16 or HPV-18, a key measure of prior exposure to these types. Methods Cross-sectional analysis of baseline visit for 296 HIV-positive MSM participating in a prospective cohort study of anal squamous intraepithelial lesions (ASIL) at a university-based research clinic. Participants completed a questionnaire detailing behaviors and medical history. Phlebotomy, anal cytology, HPV DNA testing with quantitation, and high resolution anoscopy with biopsy were performed. A pseudovirion-based neutralizing antibody (PBNA) assay was used to measure HPV-16 and HPV-18 neutralizing antibodies. Results 132/296 (45%) men were HPV-16-seropositive and 141/296 (48%) were HPV-18-seropositive. 175/296 (59%) of the men were positive for HPV-16 antibodies or DNA, and 167/296 (56%) were positive for HPV-18 antibodies or DNA. In multivariable analysis, HPV-16 seropositivity did not correlate with age, years of HIV positivity, CD4+ level or HIV viral load. Significant risk factors included HPV-16 DNA positivity with higher DNA levels (ptrend<.001) and higher number of receptive sexual partners in the last year (ptrend=.012). Conclusions A high proportion of HIV-positive MSM >26 years are DNA-negative and seronegative to HPV-16 and HPV-18 even when using a sensitive PBNA assay. Prospective studies are needed to determine the clinical- and cost-effectiveness of HPV vaccination in HIV-positive MSM > 26 years old. PMID:24231786

  17. Antibody-directed double suicide gene therapy targeting of MUC1- positive leukemia cells in vitro and in vivo.

    PubMed

    Dong, Xiao-Ya; Wang, Wen-Qian; Zhao, Yu; Li, Xu-Dong; Fang, Zhi-Gang; Lin, Dong-Jun; Xiao, Ruo-Zhi; Huang, Ren-Wei; Pan, Guang-Jin; Liu, Jia-Jun

    2013-10-01

    Our aim was to specifically transfer the cytosine deaminase (CD) and thymidine kinase (TK) genes into mucin 1 (MUC1)-positive leukemia cells by anti-MUC1 antibody directed infection of replication-defective lentivirus and to evaluate the targeted cytotoxicity of double suicide genes to leukemia. The target gene vector (containing CD and TK) and envelope (containing GFP and anti-MUC1) and packaging plasmids were cotransfected into 293T cells to produce the recombinant lentivirus. Suicide genes in virus-infected leukemia cells (U937, Jurkat, and K562) were detected by western blot. The cytotoxicity and bystander effect in vitro and the therapeutic effect in vivo were detected after treatment with the prodrugs. The results revealed that combined treatment with prodrug 5-fluorocytosine (5-FC) and ganciclovir (GCV) inhibited leukemia cell growth and caused significant bystander effect than treatment with either prodrug alone. TK/GCV treatment alone induced degeneration and cell death while the effect of CD/5-FC alone mainly caused vacuolar degeneration and necrosis. The addictive effects of combinatorial use of GCV and 5-FC mainly induced swelling of the mitochondria followed by necrosis of the leukemia cells. In vivo experiments revealed that both single and combinatorial prodrug treatments could prolong the survival time of leukemic mice. In summary, anti-MUC1 antibody directed lentiviral vector successfully transduced dual suicide genes and exerted targeted cytotoxicity against MUC1 positive leukemia cells. This targeted lentiviral dual suicide gene delivering system provides a promising approach for clinical treatment of leukemia in future. PMID:24060312

  18. Characterisation of an engineered trastuzumab IgE antibody and effector cell mechanisms targeting HER2/neu-positive tumour cells

    PubMed Central

    2010-01-01

    Trastuzumab (Herceptin®), a humanized IgG1 antibody raised against the human epidermal growth factor receptor 2 (HER2/neu), is the main antibody in clinical use against breast cancer. Pre-clinical evidence and clinical studies indicate that trastuzumab employs several anti-tumour mechanisms that most likely contribute to enhanced survival of patients with HER2/neu-positive breast carcinomas. New strategies are aimed at improving antibody-based therapeutics like trastuzumab, e.g. by enhancing antibody-mediated effector function mechanisms. Based on our previous findings that a chimaeric ovarian tumour antigen-specific IgE antibody showed greater efficacy in tumour cell killing, compared to the corresponding IgG1 antibody, we have produced an IgE homologue of trastuzumab. Trastuzumab IgE was engineered with the same light- and heavy-chain variable-regions as trastuzumab, but with an epsilon in place of the gamma-1 heavy-chain constant region. We describe the physical characterisation and ligand binding properties of the trastuzumab IgE and elucidate its potential anti-tumour activities in functional assays. Both trastuzumab and trastuzumab IgE can activate monocytic cells to kill tumour cells, but they operate by different mechanisms: trastuzumab functions in antibody-dependent cell-mediated phagocytosis (ADCP), whereas trastuzumab IgE functions in antibody-dependent cell-mediated cytotoxicity (ADCC). Trastuzumab IgE, incubated with mast cells and HER2/neu-expressing tumour cells, triggers mast cell degranulation, recruiting against cancer cells a potent immune response, characteristic of allergic reactions. Finally, in viability assays both antibodies mediate comparable levels of tumour cell growth arrest. These functional characteristics of trastuzumab IgE, some distinct from those of trastuzumab, indicate its potential to complement or improve upon the existing clinical benefits of trastuzumab. PMID:18941743

  19. Induction of high levels of antibodies recognizing the neutralizing epitope ELDKWA and the D- or K-position-mutated epitopes by candidate epitope vaccines against HIV-1.

    PubMed

    Xiao, Y; Dong, X; Chen, Y

    2000-08-01

    Monoclonal antibody 2F5 recognizing the ELDKWA epitope on HIV-1 gp41 has a significant neutralization potency against 90% of the investigated viruses of African, Asian, American, and European strains, but the antibody responses to the epitope 2F5 in HIV-1-infected individuals were very low. We attempted to induce high levels of epitope-specific antibodies to ELDKWA and its three mutated epitopes by candidate epitope vaccines. The four candidate epitope vaccines all induced strong antibody responses at dilutions from about 1:6,400 to 1:25,600. We tested the cross-reactions between these antisera and four epitope peptides. The ELDKWA-specific antisera showed strong cross-reactivity with three neutralizing-resistant mutated epitopes which contain changes in the D or K positions of the epitope sequence. Virus variants containing these changes could escape neutralization by monoclonal antibody 2F5. In immunoblotting analysis, the ELDKWA, ELDEWA, and ELEKWA epitope specific antibodies all recognized rsgp41 which confirms that the antibodies against both mutated epitopes, ELDEWA and ELEKWA, could cross-react with the native epitope on rsgp41. Although it is not clear whether the polyclonal antibodies induced by the ELDKWA epitope vaccine could neutralize the mutated viruses containing these mutated epitopes, it is conceivable that epitope vaccines based on mutated epitopes could induce strong antibody responses with predefined epitope specificity to neutralize mutated viruse containing the mutated epitope. An epitope vaccine, using different epitopes including mutated epitopes, could provide a new concept for developing a new vaccine against HIV-1. PMID:10971120

  20. International Consensus Statement on Testing and Reporting of Antineutrophil Cytoplasmic Antibodies (ANCA)

    PubMed

    Savige, J; Gillis, D; Benson, E; Davies, D; Esnault, V; Falk, R J; Hagen, E C; Jayne, D; Jennette, J C; Paspaliaris, B; Pollock, W; Pusey, C; Savage, C O; Silvestrini, R; van der Woude, F; Wieslander, J; Wiik, A

    1999-04-01

    Antineutrophil cytoplasmic antibody (ANCA) tests are used to diagnose and monitor inflammatory activity in the primary systemic small vessel vasculitides. ANCA is best demonstrated in these diseases by using a combination of indirect immunofluorescence (IIF) of normal peripheral blood neutrophils and enzyme-linked immunosorbent assays (ELISAs) that detect ANCA specific for proteinase 3 (PR3) or myeloperoxidase (MPO). For ANCA testing in "new" patients, IIF must be performed on all serum samples. Serum samples containing ANCA, any other cytoplasmic fluorescence, or an antinuclear antibody (ANA) that results in homogeneous or peripheral nuclear fluorescence then should be tested in ELISAs for PR3-ANCA and MPO-ANCA. Optimally, ELISAs for PR3-ANCA and MPO-ANCA should be performed on all serum samples. Inclusion of the most recent positive sample in the IIF or ELISA may help demonstrate a change in antibody level. Reports should use recommended terms. Any report of positive neutrophil fluorescence issued before the ELISA results are available should indicate that positive fluorescence alone is not specific for the diagnosis of Wegener granulomatosis or microscopic polyangiitis and that decisions about treatment should not be based solely on the ANCA results. PMID:10191771

  1. Combination of novel HER2-targeting antibody 1E11 with trastuzumab shows synergistic antitumor activity in HER2-positive gastric cancer.

    PubMed

    Ko, Bong-Kook; Lee, Sook-Yeon; Lee, Young-Ha; Hwang, In-Sik; Persson, Helena; Rockberg, Johan; Borrebaeck, Carl; Park, Dongeun; Kim, Kyu-Tae; Uhlen, Mathias; Lee, Jong-Seo

    2015-02-01

    The synergistic interaction of two antibodies targeting the same protein could be developed as an effective anti-cancer therapy. Human epidermal growth factor receptor 2 (HER2) is overexpressed in 20-25% of breast and gastric cancer patients, and HER2-targeted antibody therapy using trastuzumab is effective in many of these patients. Nonetheless, improving therapeutic efficacy and patient survival is important, particularly in patients with HER2-positive gastric cancer. Here, we describe the development of 1E11, a HER2-targeted humanized monoclonal antibody showing increased efficacy in a highly synergistic manner in combination with trastuzumab in the HER2-overexpressing gastric cancer cell lines NCI-N87 and OE-19. The two antibodies bind to sub-domain IV of the receptor, but have non-overlapping epitopes, allowing them to simultaneously bind HER2. Treatment with 1E11 alone induced apoptosis in HER2-positive cancer cells, and this effect was enhanced by combination treatment with trastuzumab. Combination treatment with 1E11 and trastuzumab reduced the levels of total HER2 protein and those of aberrant HER2 signaling molecules including phosphorylated HER3 and EGFR. The synergistic antitumor activity of 1E11 in combination with trastuzumab indicates that it could be a novel potent therapeutic antibody for the treatment of HER2-overexpressing gastric cancers. PMID:25306393

  2. The association of anti-annexin1 antibodies with the occurrence of skin lesions in systemic lupus erythematosus.

    PubMed

    Meng, Z; Shi, Z-R; Tan, G-Z; Yin, J; Wu, J; Mi, X-B; Wang, L

    2014-02-01

    Anti-annexin1 antibodies are associated with the subtypes of cutaneous lupus and are elevated in systemic lupus erythematosus (SLE) patients. In this study, we investigated the correlation of this antibody with the incidence of SLE skin lesions. The presence of anti-annexin1-IgG and-IgM determined by Western blot was no different among healthy controls and SLE patients with and without skin lesions. Serum levels of anti-annexin1-IgG and -IgM measured by enzyme-linked immunosorbent assay were comparable between patients with and without skin lesions, whereas anti-annexin1-IgM was lower in SLE patients than in healthy controls. Annexin1 was abundantly detected in each epidermal layer in lupus lesional skin. Additionally, anti-annexin1-IgG was higher in SLE patients with arthritis and negatively correlated with white blood cells (WBC). Anti-annexin1-IgM was higher in patients with antinuclear antibody (ANA)-positive sera, and was positively related to hemoglobin and total serum IgM. Collectively, anti-annexin1 antibodies are not related to the incidence of skin lesions in SLE, and annexin1 abundantly distributes in epidermis in lesional skin. PMID:24300781

  3. Prevalence and clinical significance of antikeratin antibodies and other serological markers in Lithuanian patients with rheumatoid arthritis

    PubMed Central

    Vasiliauskiene, L; Wiik, A; Hoier-Madsen, M

    2001-01-01

    OBJECTIVES—To assess the clinical value of several serological markers in Lithuanian patients with rheumatoid arthritis (RA) compared with control patients with rheumatic disease and age matched healthy controls.?METHODS—Serum samples from 96 patients with RA of approximately 8 years' duration, 90 rheumatic disease controls, and 37 healthy subjects were tested. Antikeratin antibody (AKA), antineutrophil cytoplasmic antibody (ANCA), and antinuclear antibody (ANA) titres were estimated by indirect immunofluorescence (IIF) and serum samples positive for ANA and ANCA were further studied by enzyme linked immunosorbent assay (ELISA). IgA and IgM rheumatoid factors (RF) were measured by ELISA.?RESULTS—A positive AKA test was highly specific for RA (diagnostic specificity 97%), being found in 44% of the patients. Although both RF tests had a higher sensitivity, they were less specific for RA. ANCA was detected in 33% of patients with RA but lacked diagnostic specificity. AKA and ANCA were associated with more erosive disease and the presence of extra-articular manifestations. Positivity for AKA, IgA RF, and ANCA was significantly associated with disease activity and worse functional capacity. However, in multiple regression analysis only positivity for AKA was significantly correlated with functional disability (p=0.0001), evaluated by the Steinbrocker functional classification, and no single marker had any relation with radiological damage.?CONCLUSION—Although AKA showed the highest disease specificity, all serological markers studied except ANA exhibited interesting associations with important clinical and paraclinical parameters of RA.?? PMID:11302867

  4. Amphibole, but not chrysotile, asbestos induces anti-nuclear autoantibodies and IL-17 in C57BL/6 mice.

    PubMed

    Ferro, Aaron; Zebedeo, Christian Nash; Davis, Chad; Ng, Kok Whei; Pfau, Jean C

    2014-01-01

    Abstract Exposure to amphibole asbestos has been associated with production of autoantibodies in mice and humans, and increases the risk of systemic autoimmune disease. However, epidemiological studies of chrysotile exposure have not indicated a similar induction of autoimmune responses. To demonstrate this difference in controlled exposures in mice, and to explore possible mechanistic explanations for the difference, C57BL/6 mice were exposed intratracheally to amphibole or chrysotile asbestos, or to saline only. Serum antinuclear antibodies (ANA), antibodies to extractable nuclear antigens (ENA), serum cytokines, and immunoglobulin isotypes were evaluated 8 months after the final treatment. The percentages of lymphocyte sub-sets were determined in the spleen and lungs. The results show that amphibole, but not chrysotile, asbestos increases the frequency of ANA/ENA in mice. Amphibole and chrysotile both increased multiple serum cytokines, but only amphibole increased IL-17. Both fibers decreased IgG1, without significant changes in other immunoglobulin isotypes. Although there were no gross changes in overall percentages of T- and B-cells in the spleen or lung, there was a significant increase in the normally rare populations of suppressor B-cells (CD19(+), CD5(+), CD1d(+)) in both the spleen and lungs of chrysotile-exposed mice. Overall, the results suggest that, while there may be an inflammatory response to both forms of asbestos, there is an autoimmune response in only the amphibole-exposed, but not the chrysotile-exposed mice. These data have critical implications in terms of screening and health outcomes of asbestos-exposed populations. PMID:24164284

  5. Delayed Development of Pulmonary Hemorrhage in a Patient with Positive Circulating Anti-Neutrophil Cytoplasmic Antibody: A Clinical Dilemma

    PubMed Central

    Imai, Toshimi; Takeda, Shin-ichi; Kawaguchi, Kazuo; Chaki, Yuko; Morishita, Yoshiyuki; Akimoto, Tetsu; Muto, Shigeaki; Kusano, Eiji

    2013-01-01

    Detection of circulating anti-neutrophil cytoplasmic antibody (ANCA) provides a powerful clue in the diagnosis of vasculitis, but the clinical interpretation of the results is difficult in some cases. Here, we describe the case of a 65-year-old man who underwent hemodialysis due to focal segmental glomerulosclerosis and abruptly developed hemoptysis 14 years after a renal biopsy. At the time of the biopsy, computed tomography (CT) showed interstitial shadows in the lungs and pleural thickening, indicating pneumoconiosis that was accompanied by tuberculosis. Circulating myeloperoxidase-ANCA (10.5–32.5 U/ml) was subsequently noted, but the significance of this observation was unclear due to the preexisting disorders in the lungs and kidneys. Potent immunosuppressive therapies were avoided because of the pulmonary lesions and decreased renal function. There were few changes noted on follow-up CT, but infiltrative shadows emerged in the bilateral lungs, consistent with hemoptysis. The hemorrhagic shadows completely disappeared shortly after initiation of steroid therapy, with normalization of the serum ANCA level. Herein, we report this case, with an emphasis on the clinical dilemma faced in deciding the appropriate treatment. The findings in the case provide deep insights into clinical management of ANCA-positive patients. PMID:24163688

  6. Lamivudine resistance leading to de novo hepatitis B infection in recipients of hepatitis B core antibody positive liver allografts.

    PubMed

    Leong, Jennifer; Coty, Patrick; Fiel, M Isabel; Chang, Charissa; Florman, Sander; Schiano, Thomas

    2014-11-01

    Most studies have shown that lamivudine (LAM) prophylaxis is sufficient to prevent hepatitis B virus (HBV) transmission in recipients of hepatitis B core antibody positive (HBcAb(+) ) allografts. However, de novo hepatitis B (DNHB) is known to occur in this patient population. Herein, we report a case series of four liver transplant recipients who developed DNHB after receiving HBcAb(+) allografts due to acquisition of LAM resistance mutations, suggesting that LAM prophylaxis may be suboptimal. A retrospective chart review was performed of all adult liver transplants performed at Mount Sinai from 2001 to 2010. A total of 79 patients received HBcAb(+) allografts for non-hepatitis B-related liver disease. Of these 79 recipients, four patients developed DNHB and were found to have documented LAM resistance. With the increasing use of HBcAb(+) donor livers, we suspect that there will also be a growing number of cases of DNHB due to acquisition of LAM resistance. We suggest that other agents, such as entecavir or tenofovir, be considered for use as prophylaxis in this patient population to decrease this risk. PMID:24107139

  7. Conjugates of folate and anti-T-cell-receptor antibodies specifically target folate-receptor-positive tumor cells for lysis.

    PubMed Central

    Kranz, D M; Patrick, T A; Brigle, K E; Spinella, M J; Roy, E J

    1995-01-01

    High-affinity folate receptors (FRs) are expressed at elevated levels on many human tumors. Bispecific antibodies that bind the FR and the T-cell receptor (TCR) mediate lysis of these tumor cells by cytotoxic T lymphocytes. In this report, conjugates that consist of folate covalently linked to anti-TCR antibodies are shown to be potent in mediating lysis of tumor cells that express either the alpha or beta isoform of the FR. Intact antibodies with an average of five folate per molecule exhibited high affinity for FR+ tumor cells but did not bind to FR- tumor cells. Lysis of FR+ cell lines could be detected at concentrations as low as 1 pM (approximately 0.1 ng/ml), which was 1/1000th the concentration required to detect binding to the FR+ cells. Various FR+ mouse tumor cell lines could be targeted with each of three different anti-TCR antibodies that were tested as conjugates. The antibodies included 1B2, a clonotypic antibody specific for the cytotoxic T cell clone 2C; KJ16, an anti-V beta 8 antibody; and 2C11, an anti-CD3 antibody. These antibodies differ in affinities by up to 100-fold, yet the cytolytic capabilities of the folate/antibody conjugates differed by no more than 10-fold. The reduced size (in comparison with bispecific antibodies) and high affinity of folate conjugates suggest that they may be useful as immunotherapeutic agents in targeting tumors that express folate receptors. PMID:7568072

  8. Clinical and genetic characteristics of GAD-antibody positive patients initially diagnosed as having type 2 diabetes

    Microsoft Academic Search

    Kazuyuki Hamaguchi; Akinori Kimura; Yoichiro Kusuda; Tsutomu Yamashita; Michio Yasunami; Megumi Takahasi; Nobuyuki Abe; Hironobu Yoshimatsu

    2004-01-01

    The present study was conducted to clarify the clinical and genetic characteristics of the diabetic patients who have antibodies to glutamic acid decarboxylase (GADab) but are diagnosed initially as type 2 diabetes because of the slow progression. Fifty-five GADab+ patients and 137 GADab? patients were recruited. The GADab+ patients were divided into two subgroups according to their antibody titers. The

  9. Detection of Toxoplasma gondii soluble antigen, SAG-1(p30), antibody and immune complex in the cerebrospinal fluid of HIV positive or negative individuals.

    PubMed

    Chaves-Borges, F A; Souza, M A; Silva, D A; Kasper, L H; Mineo, J R

    1999-01-01

    Active infection by T. gondii was evaluated by immunoassay for soluble SAG-1 (p30), the major surface antigen from T. gondii, specific antibodies and immune complexes in human cerebrospinal fluid (CSF) samples. A total of 263 samples of CSF were collected from hospitalized patients presenting neurological disorders and analyzed for antibodies to HIV. Patients were divided into two groups: HIV positive (n = 96) or HIV negative (n =167). The results of the assays showed that 45% of all samples were positive for soluble SAG-1. Toxoplasma Ag/Ab immune complexes were detected in 19% of the CSF samples and 62% were positive for T. gondii- specific IgG. A combination of these assays in the presence of clinical findings consistent with active Toxoplasma infection may predict the presence of toxoplasmic encephalitis. Moreover, detection of soluble SAG-1 in the CSF of these individuals appears consistent with active infection. PMID:10671285

  10. Skin autoreactivity in Hashimoto's thyroiditis patients without urticaria: autologous serum skin test positivity correlation with thyroid antibodies, sonographical volume and grading.

    PubMed

    Turkoglu, Zafer; Zindanci, Ilkin; Turkoglu, Ozlem; Can, Burce; Kavala, Mukaddes; Tamer, Gonca; Ulucay, Vasfiye; Akyer, Erdal

    2012-01-01

    Recent studies have shown an association between anti-thyroid antibodies and autologous serum skin test (ASST) positive urticaria patients. However, a connection between thyroid and this reliable skin test for mast cell autoreactivity, ASST, has not been reported yet. We investigated ASST in patients with Hashimoto's thyroiditis (HT) without urticaria and compared the results with laboratory and sonographical findings of HT. 154 HT patients, 100 healthy volunteers without HT as a first control group and 46 patients with multinodular goitre but without autoimmune thyroid disease as a second control group underwent testing with ASST. ASST was applied to these groups according to two criteria, first as ASST(new): autologous serum red wheal response 1.5 mm bigger than negative control; second as ASST(old): serum red wheal response 5 mm bigger than negative control accepted as positive. Free triiodothyronine (fT3), free thyroxine (fT4), thyroid-stimulating hormone (TSH), thyroid peroxidase antibody (anti-TPO) and thyroglobulin antibody (anti-Tg) levels were measured. ASST(old), ASST(new) scored positive in 51.3-60.4% of HT patients, with statistically significant differences. Thyroid volume grades were inversely proportional with ASST(old) and (new) positivity. Moderate (+) titers of anti-Tg in ASST(old) and (new) (+) cases were significantly higher than the same titers of anti-Tg in ASST(old) and (new) (-) cases. The prevalence of ASST positivity in HT patients was not affected by the following factors: gender, age at screening, laboratory measurements of thyroid function tests, anti-TPO antibodies and thyroid ultrasound (US) echogenicity. Positivity of ASST in HT has shown that there is a skin mast cell autoreactivity in HT patients independent of autoreactive chronic urticaria (ACU). PMID:22503840

  11. False-positive immunogenicity responses are caused by CD20+ B cell membrane fragments in an anti-ofatumumab antibody bridging assay.

    PubMed

    Chen, Keguan; Page, Jerry G; Schwartz, Ann M; Lee, Thomas N; DeWall, Stephen L; Sikkema, Daniel J; Wang, Catherine

    2013-08-30

    An electrochemiluminescent (ECL) bridging assay to detect anti-ofatumumab antibodies (ADA) in human serum samples was developed and validated. Using this assay format, clinical samples were first screened to identify potential ADA positive samples, which were then further tested by adding excess drug, confirming the positive signals as drug specific. However, when the method was implemented into clinical studies for ADA testing, a high positive rate was observed in the pre-dose samples collected from patients with chronic lymphocytic leukemia (CLL). Since the positive signals were not associated with ofatumumab (Ofa) treatment, and diminished after treatment, it was suspected that matrix interference might be responsible, resulting in false-positive responses. We performed a series of experimental investigations to identify, characterize, minimize or eliminate the possible false-positive responses. One possible source was identified to be CD20 (the target of Ofa) present on cell membrane fragments (CMFs). The false-positive responses caused by CD20(+) CMFs could be reduced by solid-phase immunodepletion, ultracentrifugation, or inhibited by adding another anti-CD20 antibody (rituximab). As a consequence, the ADA method was modified to minimize the matrix interference caused by CD20(+) CMFs and, then, validated for sample testing. PMID:23639298

  12. Monitoring of serum hepatitis C virus RNA level during steroid therapy for hepatitis C virus-positive antineutrophil cytoplasmic antibody-related glomerulonephritis: report of two cases

    Microsoft Academic Search

    Atsushi KURUSU; Satoshi HORIKOSHI; Satoko UEYAMA-SAKODA; Yukihiko TAKEDA; Kunimi MAEDA; Shigenobu SUZUKI; Kazuhiko FUNABIKI; Isao SHIRATO; Yasuhiko TOMINO

    2001-01-01

    We report on two adult patients with hepatitis C virus-positive antineutrophil cytoplasmic antibody-related glomerulonephritis. These patients had a renal-limited form of crescentic glomerulonephritis with high levels of serum hepatitis C virus RNA. We investigated the efficiency and transition of the serum levels of creatinine, hepatitis C virus RNA, and hepatic enzymes during corticosteroid therapy in the two patients. Corticosteroid therapy

  13. Thyroid Antibodies, Autoimmunity and Cognitive Decline: Is There a Population-Based Link?

    PubMed Central

    Napthali, Kate; Boyle, Michael; Tran, Huy; Schofield, Peter W.; Peel, Roseanne; McEvoy, Mark; Oldmeadow, Christopher; Attia, John

    2014-01-01

    Background Autoimmunity is considered an uncommon but under-recognised cause of cognitive decline. Methods Serum samples from 3,253 randomly selected subjects enrolled in the Hunter Community Study, aged 55-85 years, were assayed for thyrotropin stimulatory hormone, anti-thyroid peroxidase antibodies (TPO-Ab), anti-nuclear antibodies (ANA) and extractable nuclear antigens (ENA). Cognitive function was assessed using the Audio Recorded Cognitive Screen (ARCS) tool. Results TPO-Ab were found in 8.4% and ANA in 27.9% of the study population, of whom 3% had positive ENA findings. No relationship was found between the ARCS score and either TPO-Ab (coefficient = 0.133; 95% CI ?0.20, 0.82, p = 0.616), ANA at a low (coefficient = 1.01; 95% CI ?2.58, 0.55, p = 0.203) or a high titre (coefficient = ?0.65; 95% CI ?2.59, 1.28, p = 0.508), or ENA antibodies (coefficient = 5.12; 95% CI ?0.53, 10.77; p = 0.076). Conclusions Autoantibody findings are common in an aging population and are not associated with cognitive decline. PMID:24987403

  14. Antithyroid Antibodies Are Implicated in Epileptogenesis of Adult Patients With Epilepsy.

    PubMed

    Tsai, Meng-Han; Fu, Ting-Ying; Chen, Nai-Ching; Shih, Fu-Yuan; Lu, Yan-Ting; Cheng, Mei-Yun; Chuang, Hung-Yi; Chuang, Yao-Chung

    2015-07-01

    Antithyroid antibodies (Abs) are associated with epilepsy in steroid-responsive encephalopathy, but have been rarely studied in unselected epilepsy patients. This study aimed to characterize the prevalence and associated factors of antithyroid Abs and other auto-Abs in adult patients with epilepsy.Epilepsy patients without autoimmune disorders were surveyed for antinuclear antibody (ANA), anti-?2 glycoprotein 1 antibody (a?2GP1), anticardiolipin IgG Ab, antimicrosomal antibody (AMA), antithyroglobulin antibody (ATA), and thyroid function test.Of 319 patients, 75 (23.5%) were positive for at least 1 Ab. The most common Ab was anticardiolipin antibody (aCL) (30/319, 9.4%), followed by AMA (24/319, 7.5%), ANA (18/319, 5.6%), a?2GP1 (18/319, 6.5%), and ATA (6/319, 3.25%). Antimicrosomal Abs were significantly more frequent in patients who were female, older at disease onset, older at the time of study, and had unknown seizure etiology. The presence of aCL was significantly associated with more frequent seizures. Most patients with antithyroid Ab were female and had focal seizures with unknown etiology.The association of different auto-Abs with different factors suggests that they may have different roles in adult patients with epilepsy. Recurrent seizures and certain antiepileptic medications may cause the production of aCL. The role of antithyroid Abs in adult focal epilepsy with unknown cause, especially in females, warrants further evaluation because of the potential implications on treatment. PMID:26131823

  15. Increased detection of Sin Nombre hantavirus RNA in antibody-positive deer mice from Montana, USA: evidence of male bias in RNA viremia.

    PubMed

    Bagamian, Karoun H; Towner, Jonathan S; Mills, James N; Kuenzi, Amy J

    2013-09-01

    Hantaviruses are widespread emergent zoonotic agents that cause unapparent or limited disease in their rodent hosts, yet cause acute, often fatal pulmonary or renal infections in humans. Previous laboratory experiments with rodent reservoir hosts indicate that hantaviruses can be cleared from host blood early in the infection cycle, while sequestered long term in various host organs. Field studies of North American deer mice (Peromyscus maniculatus), the natural reservoir of Sin Nombre hantavirus, have shown that viral RNA can be transiently detected well past the early acute infection stage, but only in the minority of infected mice. Here, using a non-degenerate RT-PCR assay optimized for SNV strains known to circulate in Montana, USA, we show that viral RNA can be repeatedly detected on a monthly basis in up to 75% of antibody positive deer mice for periods up to 3-6 months. More importantly, our data show that antibody positive male deer mice are more than twice as likely to have detectable SNV RNA in their blood as antibody positive females, suggesting that SNV-infected male deer mice are more likely to shed virus and for longer periods of time. PMID:24064796

  16. Antibodies to soluble liver antigen: an additional marker in type 1 auto-immune hepatitis

    Microsoft Academic Search

    Eric Ballot; Jean Claude Homberg; Catherine Johanet

    2000-01-01

    Background\\/Aims: Auto-immune hepatitis patients are divided into two well-defined subgroups on the basis of immunoserological markers, i.e. anti-actin cable and\\/or anti-nuclear antibodies for the auto-immune hepatitis type 1, anti-liver\\/kidney microsome type 1 and\\/or anti-liver cytosol type 1 for the autoimmune hepatitis type 2. Controversial antibodies to a soluble liver antigen have been proposed as a diagnostic marker for the putative

  17. Schistosome Infection Intensity Is Inversely Related to Auto-Reactive Antibody Levels

    Microsoft Academic Search

    Francisca Mutapi; Natsuko Imai; Norman Nausch; Claire D. Bourke; Nadine Rujeni; Kate M. Mitchell; Nicholas Midzi; Mark E. J. Woolhouse; Rick M. Maizels; Takafira Mduluza

    2011-01-01

    In animal experimental models, parasitic helminth infections can protect the host from auto-immune diseases. We conducted a population-scale human study investigating the relationship between helminth parasitism and auto-reactive antibodies and the subsequent effect of anti-helminthic treatment on this relationship. Levels of antinuclear antibodies (ANA) and plasma IL-10 were measured by enzyme linked immunosorbent assay in 613 Zimbabweans (aged 2–86 years)

  18. Differences in synovial fluid cytokine levels but not in synovial tissue cell infiltrate between anti-citrullinated peptide/protein antibody-positive and –negative rheumatoid arthritis patients

    PubMed Central

    2013-01-01

    Introduction Comparative data on synovial cell infiltrate and cytokine levels in anti citrullinated peptide/protein antibody (ACPA)-positive and ACPA negative rheumatoid arthritis (RA) patients are scarce. Our aim was to analyze synovial cell infiltrate and synovial fluid (SF) levels of cytokines in patients with RA according to the presence or absence of ACPA in serum. Methods A cross-sectional study in a single center including consecutive RA patients was performed. Patients were defined as 'ACPA negative' if serum was negative to two different ACPAs [second generation commercial anti-cyclic citrullinated peptide antibodies (CCP2) and chimeric fibrin/filaggrin citrullinated antibodies]. Parallel synovial tissue (ST) biopsies and SF were obtained by knee arthroscopy. Synovial cell infiltrate and endothelial cells were analyzed by immunohistochemistry and SF levels of Th1, Th2, Th17 and pro-inflammatory cytokines by Quantibody(R) Human Array. Results A total of 83 patients underwent arthroscopy, with a mean age of 55.9?±?12 years, and mean disease duration of 45 months (interquartile range, IQR 10.8 to 122). 62% were female and 77% were ACPA positive. No significant differences were found in clinical variables, acute phase reactants, synovial cell infiltrate or lymphoid neogenesis (LN) between ACPA positive and negative patients. However ACPA positive patients had significantly higher levels of IL-1?, IL-10, IL-17 F and CC chemokine ligand 20 (CCL-20) than ACPA negative patients. Conclusions In our cohort of patients with RA no significant differences were found in synovial cell infiltrate or synovial LN according to ACPA status. However, ACPA positive patients had higher levels of T-cell derived and pro-inflammatory cytokines than ACPA negative patients. As systemic and local inflammation was similar in the two groups, these findings support a distinct synovial physiopathology. PMID:24485167

  19. A case of bilateral auricular chondritis with anti-glutamate receptor (GluR?2) antibody-positive non-herpetic acute limbic encephalitis.

    PubMed

    Nishiguchi, Ryo; Fujimoto, Takeshi; Eguchi, Katsumi; Fukuda, Yasuo; Takahashi, Yukitoshi

    2015-01-01

    A 62-year-old man experienced pain and swelling in both of his auricles. One and a half months later, he was referred to us because of a memory disturbance. A neurological examination revealed disorientation and recent memory impairment. Diffusion-weighted and fluid-attenuated inversion recovery magnetic resonance images showed high intensity and swelling lesions in the bilateral medial temporal regions. In cerebrospinal fluid, mononuclear cell counts and total protein concentration were increased, but a herpes polymerase chain reaction was negative. Thus, he was suspected to have non-herpetic acute limbic encephalitis (NHALE). In addition, relapsing polychondritis (RP) was suspected because of the bilateral auricular chondritis and type-II collagen antibody positivity in the serum. He was treated with high-dose steroid therapy (two cycles of intravenous methylprednisolone, 500?mg per day for 3 days), which was followed by oral steroid therapy. With these treatments, his symptoms, including the painful auricular swelling and memory disturbance, gradually improved. This case exhibited anti-glutamate receptor (GluR?2) antibody positivity in both serum and cerebrospinal fluid, but anti-N-methyl-D-aspartate glutamate receptor antibody (NR1 + NR2) by cell-based assay negative in cerebrospinal fluid. Although a vascular mechanism of NHALE that is associated with RP has been suggested in the literature, this autoantibody might have induced NHALE as the mechanism of neuronal damage to target neuron in our case. More studies on the pathogenesis of NHALE that is associated with RP are needed. PMID:26103811

  20. Protection from clinical disease against three highly virulent strains of Newcastle disease virus after in ovo application of an antibody-antigen complex vaccine in maternal antibody-positive chickens.

    PubMed

    Kapczynski, Darrell R; Martin, Alison; Haddad, Eid E; King, Daniel J

    2012-09-01

    Worldwide, Newcastle disease (ND) remains one of the most economically important diseases of poultry. Current vaccination strategies for commercial poultry include the use of inactivated and live ND vaccines that typically induce protection against virulent field viruses. Here, we tested the efficacy of an antigen-antibody complex (AAC) ND vaccine delivered in ovo. Commercial maternal antibody-positive broiler chickens (Gallus domesticus) were vaccinated in ovo with an AAC vaccine composed of live B1-LaSota Newcastle disease virus (NDV) complexed with NDV-specific antiserum, and then they were challenged at weekly intervals after hatch. Challenge viruses included three exotic ND disease (END) viruses: the neurotropic strain Texas GB NDV-92-01 (TxGB) and two viscerotropic isolates, one isolate from the 2002-2003 outbreak in California (California 2002 isolate S212676 [CA]) and the other isolate from a 1997 END outbreak in South Korea (South Korea 94-147 [SK]). Results demonstrate that maternal antibody was able to provide approximately 50% protection in either vaccinated or control chickens at 7 days of age after TxGB challenge. However, with challenge at > or = 14 days, most control birds died, whereas all AAC-vaccinated birds were protected. Challenge with the CA or SK viruses in chickens at 28 days of age resulted in 100% protection of vaccinated birds, whereas all control birds died. In addition, AAC-vaccinated birds displayed decreased incidence of viral shedding in oral and cloacal swabs than control birds. Antibody titers were significantly (P < 0.05) higher in vaccinated chickens, as determined by enzyme-linked immunosorbent assay and hemagglutinin-inhibition tests, than in nonvaccinated controls. Together, these results demonstrate the efficacy of AAC vaccines delivered in ovo to protect commercial poultry. PMID:23050473

  1. Humanization of high affinity anti-HBs antibody by using human consensus sequence and modification of selected minimal positional template and packing residues.

    PubMed

    Tiwari, Ashutosh; Khanna, Navin; Acharya, Subrat K; Sinha, Subrata

    2009-04-14

    We had earlier reported the construction and characterization of a high affinity recombinant scFv generated from a potential neutralizing mouse monoclonal antibody against the Hepatitis B surface antigen. In this report we describe the humanization of this scFv by grafting its antigen binding site onto framework of the human consensus sequence of highest similarity. We have used molecular modeling to alter not only the clearly permissible residues but also several minimal positional template and V(H)/V(L) interface residues. The humanized scFv retains the binding characteristic of the mouse monoclonal even under conditions that usually destabilize antigen antibody interactions. This high affinity humanized scFv provides a basis for the development of prophylactic/therapeutic molecules. PMID:19428851

  2. High levels of soluble CTLA-4 are present in anti-mitochondrial antibody positive, but not in antibody negative patients with primary biliary cirrhosis.

    PubMed

    Saverino, Daniele; Pesce, Giampaola; Antola, Princey; Porcelli, Brunetta; Brusca, Ignazio; Villalta, Danilo; Tampoia, Marilina; Tozzoli, Renato; Tonutti, Elio; Alessio, Maria Grazia; Bagnasco, Marcello; Bizzaro, Nicola

    2014-01-01

    Primary biliary cirrhosis (PBC) is a chronic autoimmune cholestatic liver disease frequently characterized by anti-mitochondrial autoantibodies (AMA). A minority of patients are AMA-negative. Cytotoxic-T-Lymphocyte-Antigen-4 (CTLA-4) is a surface molecule expressed on activated T-cells delivering a critical negative immunoregulatory signal. A soluble form of CTLA-4 (sCTLA-4) has been detected at high concentrations in several autoimmune diseases, and its possible functional meaning has been suggested. We aimed to evaluate sCTLA-4 concentration in sera of patients with PBC and to correlate it to immunological abnormalities associated with the disease. Blood samples were collected from 82 PBC-patients diagnosed according to international criteria (44 AMA-positive/MIT3-positive and 38 AMA-negative-MIT3-negative), and 65 controls. sCTLA-4 levels were evaluated by ELISA and Western blot. Increased sCTLA-4 concentrations were found in all AMA-positive PBC-patients, but in none of the AMA-negative ones, nor in normal controls or in controls with unrelated liver diseases. sCTLA-4 presence was associated with autoantibodies against MIT3, but not with nuclear autoantibodies (sp100, gp210). This is the first study to demonstrate that levels of sCTLA-4 are elevated in sera of PBC patients. However, they are clearly restricted to patients with AMA positivity, suggesting an immunological difference with respect to AMA-negative ones. PMID:25383768

  3. The Leukocyte Antibody Prevalence Study-II (LAPS-II): a retrospective cohort study of transfusion-related acute lung injury in recipients of high-plasma-volume human leukocyte antigen antibody–positive or –negative components

    PubMed Central

    Kleinman, Steven H.; Triulzi, Darrell J.; Murphy, Edward L.; Carey, Patricia M.; Gottschall, Jerome L.; Roback, John D.; Carrick, Danielle; Mathew, Sunitha; Wright, David J.; Cable, Ritchard; Ness, Paul; Gajic, Ognjen; Hubmayr, Rolf D.; Looney, Mark R.; Kakaiya, Ram M.

    2013-01-01

    BACKGROUND We used a multicenter retrospective cohort study design to evaluate whether human leukocyte antigen (HLA) antibody donor screening would reduce the risk of transfusion-related acute lung injury (TRALI) or possible TRALI. STUDY DESIGN AND METHODS In the Leukocyte Antibody Prevalence Study-II (LAPS-II), we evaluated pulmonary outcomes in recipients of 2596 plasma-rich blood components (transfusable plasma and plateletpheresis) sent to participating hospitals; half of the components were collected from anti-HLA–positive donors (study arm) and half from anti-HLA–negative donors (control arm) matched by sex, parity, and blood center. A staged medical record review process was used. Final recipient diagnosis was based on case review by a blinded expert panel of pulmonary or critical care physicians. RESULTS TRALI incidence was 0.59% (seven cases) in study arm recipients versus 0.16% (two cases) in control arm recipients for an odds ratio (OR) of 3.6 (95% confidence interval [CI], 0.7–17.4; p = 0.10). For possible TRALI cases (nine study arm, eight control arm), the OR was 1.2 (95% CI, 0.4–3.0; p = 0.81), and for TRALI and possible TRALI aggregated together, it was 1.7 (95% CI, 0.7–3.7; p = 0.24). Transfusion-associated circulatory overload incidence was identical in the two arms (1.17 and 1.22%, respectively; OR, 1.0; p = 1.0). CONCLUSIONS TRALI incidence in recipients of anti-HLA–positive components was relatively low for a look-back study (1 in 170) and was higher than in the control arm, but did not reach significance. Based on this trend, the data are consistent with the likelihood that TRALI risk is decreased by selecting high-volume plasma components for transfusion from donors at low risk of having HLA antibodies. PMID:21446938

  4. Subclinical Sjögren's syndrome and anti-Ro/SSA-positive autoimmune fatigue syndrome in children.

    PubMed

    Itoh, Y; Imai, T; Fujino, O; Igarashi, T; Fukunaga, Y

    2002-09-01

    Abstract Although Sjögren's syndrome (SS) is quite rare among children, subclinical conditions without any sicca symptoms have been reported. This condition is characterized by nonspecific rheumatic symptoms and histopathological findings in salivary glands which are equivalent to SS. Many children with subclinical SS are positive for anti-Ro/SSA. On the other hand, autoimmune fatigue syndrome (AIFS) is characterized by chronic nonspecific complaints and positive antinuclear antibodies, with or without fulfilling the criteria for chronic fatigue syndrome. Although a novel autoantibody against a 62?kD nuclear protein (anti-Sa) is detected in about 40% of AIFS patients, few marker antibodies for autoimmune diseases, such as anti-DNA, anti-Sm, anti-U1-ribonucleoprotein (RNP), or anticardiolipin, are found in AIFS patients. In this study, however, anti-Ro/SSA was detected in sera from 8 out of 122 AIFS patients. Seven of the 8 anti-Ro/SSA-positive patients were female. All 8 patients had fatigue and low-grade fever, but none complained of xerosis. Western immunoblot analysis revealed that 7 sera reacted with Ro52, and that none was positive for anti-La/SSB or anti-Sa. Two of the 8 patients had histories of recurrent parotitis. Lip biopsies showed mild chronic inflammation compatible with subclinical SS in these 2 patients, although the other 6 patients had no abnormal histopathology. Thus, at least some anti-Ro/SSA-positive patients could be diagnosed as having SS. PMID:24387058

  5. Different amounts of protein-bound citrulline and homocitrulline in foot joint tissues of a patient with anti-citrullinated protein antibody positive erosive rheumatoid arthritis

    PubMed Central

    2013-01-01

    Background Antibodies binding to citrullinated proteins are a frequent finding in rheumatoid arthritis patients and may precede the onset of clinical symptoms several years. The antibodies are a predisposing factor for bone erosions but their origin is unknown. In this study we analyze in detail the levels of protein bound citrulline and homocitrulline in several tissue samples of a single erosive arthritic surgery patient. Methods Serum antibodies binding to CCP, MCV and citrulline- or homocitrulline-containing type I and II collagen carboxytelopeptides were measured. Tissue samples of a single RA patient, taken in two separate operations performed with two-year time span were hydrolyzed and analyzed for citrulline and homocitrulline content by HPLC. Results Protein-bound citrulline and homocitrulline were found in several joint tissues of a RA patient with ACPA-positive erosive disease. The amount of homocitrulline stayed relatively constant between the different tissues. The amount of citrulline in erosive tissue was 3-times higher than in non-erosive tissue in the first operation. In the samples of the second operation 3-4-times higher mean amounts of citrulline were found in two out of the six tissues investigated. Conclusions Homocitrulline is present in rheumatoid nodule together with citrulline. There is more variation in the amount of citrulline than in the amount of homocitrulline between the tissues. The tissue sample containing the most citrulline was the most erosive. PMID:24060405

  6. Accumulation of mature B cells in the inflamed muscle tissue of a patient with anti-155/140 antibody-positive juvenile dermatomyositis.

    PubMed

    Yokoyama, Tadafumi; Shimizu, Masaki; Ishikawa, Sayaka; Ueno, Kazuyuki; Hamaguchi, Yasuhito; Takehara, Kazuhiko; Yachie, Akihiro

    2013-01-01

    We report the first case of a Japanese patient with anti-155/140 antibody-positive juvenile dermatomyositis (JDM). Her clinical features included severe cutaneous involvement. Serum B cell-activating factor levels were significantly increased. Mature class-switched memory B cells accumulated in inflamed muscle tissue but decreased in peripheral blood. These findings indicate that loss of B cell tolerance and accumulation of mature B cells in inflamed muscle tissue play an important role in the pathogenesis of JDM. PMID:22454192

  7. Levetiracetam effect on adult-onset temporal lobe epilepsy with positive voltage-gated potassium channel antibody.

    PubMed

    Wang, Wei; Zheng, Hong-Bo; Xiao, Jing; Chen, Lei; Zhou, Dong

    2015-01-01

    Temporal lobe epilepsy is considered to be the most frequent of all epileptic syndromes. Recently, several retrospective studies suggest that limbic encephalitis (LE) may be a cause for adult onset unexplained seizure disorders in patients. This report describes two cases of adult onset epilepsy with voltage-gated potassium channel antibodies (VGKC-abs)-associated LE that responded well to levetiracetam (LEV). As demonstrated by these two cases and reviewing previous reports, we propose that the therapeutic regimen for VGKC-abs associated seizures still needs to be determined and LEV may be effective in treating this kind of disorders. PMID:25321075

  8. Anti-Malondialdehyde Antibodies in MRL+\\/+ Mice Treated with Trichloroethene and Dichloroacetyl Chloride: Possible Role of Lipid Peroxidation in Autoimmunity

    Microsoft Academic Search

    M. Firoze Khan; Xiaohong Wu; G. A. S. Ansari

    2001-01-01

    Trichloroethene (TCE) and one of its metabolites dichloroacetyl chloride (DCAC) are known to induce\\/accelerate autoimmune (AI) response in MRL+\\/+ mice as evident from anti-nuclear, anti-ssDNA, anti-cardiolipin, and DCAC-specific antibodies in the serum (Khan et al., Toxicol. Appl. Pharmacol. 134, 155–160, 1995). In the present study, we measured anti-malondialdehyde antibodies (AMDA) in the serum of TCE- or DCAC-treated mice in order

  9. Anti-neutrophil cytoplasmic antibody positivity in five children with systemic lupus erythematosus--what is the importance of this finding?

    PubMed

    Bobek, Dubravka; Vukovi?, Jurica; Malenica, Branko; Bojani?, Katarina; Rukavina, Iva; Jeluši?, Marija

    2014-12-01

    Juvenile systemic lupus erythematosus (JSLE) is a systemic autoimmune chronic disease that can affect any part of the body. It is characterized by the formation of antibodies against nuclear antigens. Vasculitis may be found in SLE, but it scarcely complies with anti-neutrophil cytoplasmic antibodies (ANCA)-associated vasculitis (AAV) criteria. We report five cases of severe JSLE associated with AAV diagnosed between 1991 and 2013 in three university-based tertiary care centers. The patients (3 girls and 2 boys, aged 12 to 17) presented with a severe clinical picture and the following features: cytopenia (n=5), autoimmune hepatitis (n=3), lupus nephritis (n=1), pancreatitis (n=1), secondary antiphospholipid syndrome (n=2), impending respiratory failure (n=2), and gastrointestinal bleeding (n=1). All patients were proteinase 3 (PR3) ANCA positive, while two of them were myeloperoxidase (MPO) and PR3 ANCAs positive at the same time. They were treated with corticosteroids and immunosuppressive drugs. Remission of the disease was achieved in three patients. The course of the disease was worsening in two patients and we included rituximab (anti-CD20) in therapy. All of our patients presented as the most severe SLE patients, who must be diagnosed as soon as possible and treated very intensively. Since the comorbidity of JSLE and AAV occurs very rarely in children, presentation of such patients, their clinical pictures, treatment, and the course of the diseases are experiences that can be of great help. PMID:25580781

  10. Clinical and genetic characteristics of GAD-antibody positive patients initially diagnosed as having type 2 diabetes.

    PubMed

    Hamaguchi, Kazuyuki; Kimura, Akinori; Kusuda, Yoichiro; Yamashita, Tsutomu; Yasunami, Michio; Takahasi, Megumi; Abe, Nobuyuki; Yoshimatsu, Hironobu

    2004-11-01

    The present study was conducted to clarify the clinical and genetic characteristics of the diabetic patients who have antibodies to glutamic acid decarboxylase (GADab) but are diagnosed initially as type 2 diabetes because of the slow progression. Fifty-five GADab+ patients and 137 GADab- patients were recruited. The GADab+ patients were divided into two subgroups according to their antibody titers. The high-titer subgroup (Ab > or = 20 U/ml) had lower urinary C-peptide concentrations, and was assigned insulin therapy more often than the GADab- patients. In contrast to the high-titer subgroup, clinical parameters in the low-titer subgroup were similar to the GADab- diabetic patients. The urinary C-peptide levels correlated negatively with the GADab titer in the GADab+ patients. Analysis of type 1 diabetes-susceptible HLA alleles revealed high frequencies of the B54 and DRB1*0405 allele, but not the B61 and DRB1*0901 alleles, in the high-titer subgroup, whereas the frequency of the protective DRB1*1502 allele was decreased. The GADab+ patients with the B54 allele had higher GADab titers and lower urinary C-peptide excretion than patients without this allele. These data indicated that patients with a high-GADab titer share the autoimmune background characteristic of type 1 diabetes. PMID:15533584

  11. Impact of a single nucleotide polymorphism upstream of the IL28B gene in patients positive for anti-HCV antibody in an HCV hyperendemic area in Japan.

    PubMed

    Oda, Kohei; Uto, Hirofumi; Kumagai, Kotaro; Ido, Akio; Kusumoto, Kazunori; Shimoda, Kazuya; Hayashi, Katsuhiro; Stuver, Sherri O; Tanaka, Yasuhito; Nishida, Nao; Tokunaga, Katsushi; Tsubouchi, Hirohito

    2014-11-01

    The influence of genetic variation at the interleukin-28B (IL28B) locus on the natural course of hepatitis C virus (HCV) infection has not been fully investigated. The goal of this study was to examine whether an IL28B polymorphism (rs8099917) is associated with natural clearance of HCV and with disease parameters of HCV infection in an HCV hyperendemic area of Japan. The patients were 502 anti-HCV antibody-positive residents who participated in liver disease screening program from 2002 to 2004. Patients who underwent interferon-based therapy or had hepatocellular carcinoma were excluded. Of these patients, 149 were negative for HCV RNA (prior infection) and 353 were positive for HCV RNA or HCV core antigen (HCV carriers). In multivariate analysis, the IL28B TT genotype was a predictor for prior HCV infection. In addition, nine of the patients with prior HCV infection were positive for anti-HCV antibody with positive for HCV core antigen or HCV RNA before 2001, and these nine patients all had the IL28B TT genotype. Furthermore, the IL28B TT genotype was associated independently with higher HCV core antigen levels in HCV carriers. In contrast, the IL28B genotype did not affect the biochemical markers, such as alanine aminotransferase, hepatic fibrosis markers, and ?-fetoprotein, and the degree of hepatic fibrosis assessed by transient elastography in HCV carriers. We concluded that IL28B polymorphism (TT genotype) is associated with spontaneous clearance of HCV and conversely with high viral loads in HCV carriers. In contrast, the IL28B genotype does not affect disease progression such as hepatic fibrosis. PMID:25100136

  12. Multiple cardiac thrombi and thromboembolism in a heparin-induced thrombocytopenia antibody-positive patient with heart failure.

    PubMed

    Wake, Ryotaro; Muro, Takashi; Hozumi, Takeshi; Matsumoto, Ryo; Kataoka, Toru; Nakamura, Yasuhiro; Takemoto, Yasuhiko; Takagi, Masahiko; Suehiro, Shigefumi; Yoshiyama, Minoru

    2008-11-01

    A 49-year-old Japanese man presented with orthopnea and edema in both legs. He was diagnosed with congestive heart failure with triple-vessel coronary artery disease. Low antithrombin and left ventricular systolic dysfunction were possible causes of his hypercoagulable state. Echocardiography revealed thrombi in the left ventricle and left trium, poor left ventricular contractility, and a normal mitral valve. Electrocardiogram revealed normal sinus rhythm. We found small infarctions of the brain and spleen in the computed tomography. The heparin treatment of cardiac thrombi is useless because the patient had heparin-induced thrombocytopenia antibody. We removed thrombi in the left ventricle and left atrium by thrombectomy and performed coronary artery bypass graft. Warfarin was administered for anticoagulation. He recovered completely and is now doing well. Our experience indicates that poor cardiac function can together cause multiple cardiac thrombi and subsequent thromboembolism without mitral stenosis or atrial fibrillation. PMID:19091284

  13. Nuclear Energy. It is not a solution, it is a problem The Mediterranean Antinuclear Watch (MANW) is a non -

    E-print Network

    Nuclear Energy. It is not a solution, it is a problem #12;The Mediterranean Antinuclear Watch (MANW - called "peaceful use" of nuclear energy as well as the production and proliferation of nuclear weapons pose. #12;Nuclear energy renaissance Twenty two years after the accident in Chernobyl NPP. Energy

  14. Nanoparticle mediated drug delivery of rolipram to tyrosine kinase B positive cells in the inner ear with targeting peptides and agonistic antibodies

    PubMed Central

    Glueckert, Rudolf; Pritz, Christian O.; Roy, Soumen; Dudas, Jozsef; Schrott-Fischer, Anneliese

    2015-01-01

    Aim: Systemic pharmacotherapies have limitation due to blood-labyrinth barrier, so local delivery via the round window membrane opens a path for effective treatment. Multifunctional nanoparticle (NP)-mediated cell specific drug delivery may enhance efficacy and reduce side effects. Different NPs with ligands to target TrkB receptor were tested. Distribution, uptake mechanisms, trafficking, and bioefficacy of drug release of rolipram loaded NPs were evaluated. Methods: We tested lipid based nanocapsules (LNCs), Quantum Dot, silica NPs with surface modification by peptides mimicking TrkB or TrkB activating antibodies. Bioefficacy of drug release was tested with rolipram loaded LNCs to prevent cisplatin-induced apoptosis. We established different cell culture models with SH-SY-5Y and inner ear derived cell lines and used neonatal and adult mouse explants. Uptake and trafficking was evaluated with FACS and confocal as well as transmission electron microscopy. Results: Plain NPs show some selectivity in uptake related to the in vitro system properties, carrier material, and NP size. Some peptide ligands provide enhanced targeted uptake to neuronal cells but failed to show this in cell cultures. Agonistic antibodies linked to silica NPs showed TrkB activation and enhanced binding to inner ear derived cells. Rolipram loaded LNCs proved as effective carriers to prevent cisplatin-induced apoptosis. Discussion: Most NPs with targeting ligands showed limited effects to enhance uptake. NP aggregation and unspecific binding may change uptake mechanisms and impair endocytosis by an overload of NPs. This may affect survival signaling. NPs with antibodies activate survival signaling and show effective binding to TrkB positive cells but needs further optimization for specific internalization. Bioefficiacy of rolipram release confirms LNCs as encouraging vectors for drug delivery of lipophilic agents to the inner ear with ideal release characteristics independent of endocytosis. PMID:26042029

  15. Anti-CD20 single chain variable antibody fragment-apolipoprotein A-I chimera containing nanodisks promote targeted bioactive agent delivery to CD20-positive lymphomas.

    PubMed

    Crosby, Natasha M; Ghosh, Mistuni; Su, Betty; Beckstead, Jennifer A; Kamei, Ayako; Simonsen, Jens B; Luo, Bing; Gordon, Leo I; Forte, Trudy M; Ryan, Robert O

    2015-08-01

    A fusion protein comprising an ?-CD20 single chain variable fragment (scFv) antibody, a spacer peptide, and human apolipoprotein (apo) A-I was constructed and expressed in Escherichia coli. The lipid interaction properties intrinsic to apoA-I as well as the antigen recognition properties of the scFv were retained by the chimera. scFv•apoA-I was formulated into nanoscale reconstituted high-density lipoprotein particles (termed nanodisks; ND) and incubated with cultured cells. ?-CD20 scFv•apoA-I ND bound to CD20-positive non-Hodgkins lymphoma (NHL) cells (Ramos and Granta) but not to CD20-negative T lymphocytes (i.e., Jurkat). Binding to NHL cells was partially inhibited by pre-incubation with rituximab, a monoclonal antibody directed against CD20. Confocal fluorescence microscopy analysis of Granta cells following incubation with ?-CD20 scFv•apoA-I ND formulated with the intrinsically fluorescent hydrophobic polyphenol, curcumin, revealed ?-CD20 scFv•apoA-I localizes to the cell surface, while curcumin off-loads and gains entry to the cell. Compared to control incubations, viability of cultured NHL cells was decreased upon incubation with ?-CD20 scFv•apoA-I ND harboring curcumin. Thus, formulation of curcumin ND with ?-CD20 scFv•apoA-I as the scaffold component confers cell targeting and enhanced bioactive agent delivery, providing a strategy to minimize toxicity associated with chemotherapeutic agents. PMID:25994015

  16. Staining Pattern Classification of Antinuclear Autoantibodies Based on Block Segmentation in Indirect Immunofluorescence Images

    PubMed Central

    Li, Jiaqian; Tseng, Kuo-Kun; Hsieh, Zu Yi; Yang, Ching Wen; Huang, Huang-Nan

    2014-01-01

    Indirect immunofluorescence based on HEp-2 cell substrate is the most commonly used staining method for antinuclear autoantibodies associated with different types of autoimmune pathologies. The aim of this paper is to design an automatic system to identify the staining patterns based on block segmentation compared to the cell segmentation most used in previous research. Various feature descriptors and classifiers are tested and compared in the classification of the staining pattern of blocks and it is found that the technique of the combination of the local binary pattern and the k-nearest neighbor algorithm achieve the best performance. Relying on the results of block pattern classification, experiments on the whole images show that classifier fusion rules are able to identify the staining patterns of the whole well (specimen image) with a total accuracy of about 94.62%. PMID:25474260

  17. Limitations of a single-point evaluation of anti-MDA5 antibody, ferritin, and IL-18 in predicting the prognosis of interstitial lung disease with anti-MDA5 antibody-positive dermatomyositis.

    PubMed

    Muro, Yoshinao; Sugiura, Kazumitsu; Akiyama, Masashi

    2013-03-01

    Autoantibodies against melanoma differentiation-associated gene 5 (MDA5) are important serological markers in dermatomyositis (DM) with rapidly progressive interstitial lung disease (ILD). Recent studies noted that anti-MDA5 antibody (anti-MDA5ab), ferritin, and IL-18 are useful biomarkers for evaluating the responses to treatment and the status of ILD in anti-MDA5ab-positive DM. In this study, we further studied the importance of anti-MDA5ab levels and of ferritin and IL-18 concentrations in our patients. These biomarkers could be sometimes useful for evaluating ILD status and/or predicting the prognosis in patients with anti-MDA5ab-positive DM with several exceptional cases. A single-point evaluation of anti-MDA5ab levels and of ferritin and IL-18 concentrations has limitations in predicting the prognosis of ILD with DM. We consider that the timing of initial therapy and the anti-MDA5ab isotype, in addition to the patient's age, are also crucial factors for predicting the prognosis. PMID:23250474

  18. A Prospective Open-label Pilot Study of Fluvastatin on Pro-inflammatory and Pro-thrombotic Biomarkers in Antiphospholipid Antibody Positive Patients

    PubMed Central

    Erkan, Doruk; Willis, Rohan; Murthy, Vijaya L.; Basra, Gurjot; Vega, JoAnn; Ruiz Limón, Patricia; Carrera, Ana Laura; Papalardo, Elizabeth; Martínez-Martínez, Laura Aline; González, Emilio B.; Pierangeli, Silvia S.

    2014-01-01

    Objective: To determine if pro-inflammatory and pro-thrombotic biomarkers are differentially upregulated in persistently antiphospholipid antibody (aPL)-positive patients, and to examine the effects of fluvastatin on these biomarkers. Methods: Four groups of patients (age 18-65) were recruited: a) Primary Antiphospholipid Syndrome (PAPS); b) Systemic Lupus Erythematosus (SLE) with APS (SLE/APS); c) Persistent aPL positivity without SLE or APS (Primary aPL); and d) Persistent aPL positivity with SLE but no APS (SLE/aPL). The frequency-matched control group, used for baseline data comparison, was identified from a databank of healthy persons. Patients received fluvastatin 40 mg daily for three months. At three months, patients stopped the study medication and they were followed for another three months. Blood samples for 12 pro-inflammatory and pro-thrombotic biomarkers were collected monthly for six months. Results: Based on the comparison of the baseline samples of 41 aPL-positive patients with 30 healthy controls, 9/12 (75%) biomarkers (interleukin [IL]-6, IL1?, vascular endothelial growth factor [VEGF], tumor necrosis factor [TNF]-??, interferon [IFN]-?, inducible protein-10 [IP10], soluble CD40 ligand [sCD40L], soluble tissue factor [sTF], and intracellular cellular adhesion molecule [ICAM]-1) were significantly elevated. Twenty-four patients completed the study; fluvastatin significantly and reversibly reduced the levels of 6/12 (50%) biomarkers (IL1?, VEGF, TNF?, IP10, sCD40L, and sTF). Conclusion: Our prospective mechanistic study demonstrates that pro-inflammatory and pro-thrombotic biomarkers, which are differentially upregulated in persistently aPL-positive patients, can be reversibly reduced by fluvastatin. Thus, statin-induced modulation of the aPL effects on target cells can be a valuable future approach in the management of aPL-positive patients. PMID:23933625

  19. Positive correlation between replication rate and pathotype of Marek’s disease virus strains in maternal antibody negative chickens

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Pathotyping of new field strains of MDV requires both a long period of time and a large number of birds. Confirming a positive correlation of virus replication and pathotype may lead to faster and cheaper alternative pathotyping methods or as a screening assay for choosing isolates to be pathotyped....

  20. Trastuzumab-based treatment of HER2-positive breast cancer: an antibody-dependent cellular cytotoxicity mechanism?

    Microsoft Academic Search

    L Arnould; M Gelly; F Penault-Llorca; L Benoit; F Bonnetain; C Migeon; V Cabaret; V Fermeaux; P Bertheau; J Garnier; J-F Jeannin; B Coudert

    2006-01-01

    This study evaluated by immunohistochemistry (IHC) immune cell response during neoadjuvant primary systemic therapy (PST) with trastuzumab in patients with HER2-positive primary breast cancer. In all, 23 patients with IHC 3+ primary breast cancer were treated with trastuzumab plus docetaxel. Pathological complete and partial responses were documented for nine (39%) and 14 (61%) patients, respectively. Case-matched controls comprised patients treated

  1. Profiling anti-cyclic citrullinated peptide antibodies in patients with juvenile idiopathic arthritis

    PubMed Central

    2012-01-01

    Background Anti-citrullinated protein/peptide antibodies (ACPA), have high specificity for rheumatoid arthritis (RA). Some children with juvenile idiopathic arthritis (JIA), phenotypically resemble RA and test positive for rheumatoid factor (RF) a characteristic biomarker of RA. We investigated the prevalence of ACPA and its relationship to other serologic markers associated with RA in a well-characterized JIA cohort. Methods Cases were 334 children with JIA, 30 of whom had RF + polyarticular JIA. Sera from all cases and 50 healthy pediatric controls were investigated by ELISA at a single time point for anti-cyclic citrullinated peptide (anti-CCP) IgG, RF IgM, IgA and IgG, anti-RA33 IgG, and antinuclear antibodies (ANA). Comparisons between cases and controls were made using Chi-square or Fisher exact tests and T-tests. Results The prevalence of RF was 8% among controls, and 12% among cases (ns). The prevalence of ACPA was 2% in controls and 14.3% in cases (OR 8.2, p <0.01). Children who were ACPA-positive and RF-negative (n = 23) had a significantly earlier onset-age (4.6 years vs. 12.1 years, p <0.00001) and had fewer HLA-DRB1 shared epitope alleles than those positive for both RF and ACPA (n = 25). Prevalence of anti-RA33 was not different between cases and controls. Conclusions ACPAs are detectable in 14% of children with JIA. Children with positive ACPA but negative RF are frequent, and may define a distinct subset of children with JIA. ACPA testing should be included in the classification of JIA. PMID:22931121

  2. Antibodies to human gastric epithelial cells and heat shock protein 60 in Helicobacter pylori positive mucosa associated lymphoid tissue lymphoma

    Microsoft Academic Search

    Y Kawahara; K Yokota; M Mizuno; N Yunoki; T Uesu; H Okada; K Kobayashi; Y Hirai; K Oguma; T Tsuji

    1999-01-01

    BACKGROUNDDevelopment of gastric mucosa associated lymphoid tissue (MALT) lymphoma is thought to be closely associated with host immune reactions toHelicobacter pylori.AIMTo investigate humoral immune responses in patients with MALT lymphoma to antigens shared by H pylori and human gastric epithelial cells.METHODSSera were obtained from H pylori positive patients with MALT lymphoma (n = 11) or other gastroduodenal diseases (peptic ulcer,

  3. Anti-Peptidoglycan Antibodies and Fc? Receptors Are the Key Mediators of Inflammation in Gram-Positive Sepsis

    PubMed Central

    Sun, Dawei; Raisley, Brent; Langer, Marybeth; Iyer, Janaki K.; Vedham, Vidya; Ballard, Jimmy L.; James, Judith A.; Metcalf, Jordan

    2012-01-01

    Gram-positive bacteria are an important public health problem, but it is unclear how they cause systemic inflammation in sepsis. Our previous work showed that peptidoglycan (PGN) induced proinflammatory cytokines in human cells by binding to an unknown extracellular receptor, followed by phagocytosis leading to the generation of NOD ligands. In this study, we used flow cytometry to identify host factors that supported PGN binding to immune cells. PGN binding required plasma, and plasma from all tested healthy donors contained IgG recognizing PGN. Plasma depleted of IgG or of anti-PGN Abs did not support PGN binding or PGN-triggered cytokine production. Adding back intact but not F(ab?)2 IgG restored binding and cytokine production. Transfection of HEK293 cells with Fc?RIIA enabled PGN binding and phagocytosis. These data establish a key role for anti-PGN IgG and Fc?Rs in supporting inflammation to a major structural element of Gram-positive bacteria and suggest that anti-PGN IgG contributes to human pathology in Gram-positive sepsis. PMID:22815288

  4. Lack of Chromatin and Nuclear Fragmentation In Vivo Impairs the Production of Lupus Anti-Nuclear Antibodies1

    Microsoft Academic Search

    Lorenza Frisoni; Lenese McPhie; Sun-Ah Kang; Marc Monestier; Michael Madaio; Minoru Satoh; Roberto Caricchio

    2007-01-01

    Nuclear autoantigens in systemic lupus erythematosus are thought to derive primarily from apoptotic cells, yet there is no direct evidence that interfering with apoptosis impairs the generation of lupus autoantibodies. Here we use a mouse model that lacks the endonuclease caspase-activated DNase (CAD), resulting in an absence of chromatin and nuclear fragmentation during apoptotic cell death. We show that in

  5. A strong association between thyrotropin receptor-blocking antibody-positive atrophic autoimmune thyroiditis and HLA-DR8 and HLA-DQB1 0302 in Koreans

    SciTech Connect

    Cho, Bo Youn; Chung, Jae Hoon; Lee, Hong Kyu; Koh, Chang-Soon; Lee, Jung-Bin (Seoul National Univ. College of Medicine, Seoul (Korea, Republic of)); Shong, Young Kee (Univ. of Ulsan College of Medicine Ulsan (Korea, Republic of)); Han, Hoon (Catholic Univ. Medical College, Seoul (Korea, Republic of)); Chang, Youn Bok (Hallym Univ. College of Medicine, Seoul (Korea, Republic of))

    1993-09-01

    The authors investigated whether the associations between HLA alleles of patients with autoimmune hypothyroidism varied according to the presence or absence of TSH receptor-blocking antibody (TRBab). They analyzed the HLA-A, -B, -C, and -DR antigens by serotyping and the DQA1 and DQB1 genes using both enzymatic DNA amplification and sequence-specific oligonucleotide hybridizations. The patient population consisted of 47 Korean patients with atrophic autoimmune thyroiditis and 62 patients with goitrous autoimmune thyroiditis. The antigen frequency of HLA-DR8 was significantly increased in 23 atrophic autoimmune thyroiditis patients that were positive for TSH binding inhibitor immunoglobulin (TBII) compared to 136 controls [52% vs. 16%; x[sup 2] = 13.1; Pc (corrected P value) = 0.003]. This relative risk was 5.7; the etiological fraction was 0.43. HLA-DQB1*0302 was also increased in patients with TBII-positive atrophic autoimmune thyroiditis (24% vs. 7%; x[sup 2] = 11.2; Pc = 0.012; relative risk = 4.4; etiological fraction = 0.19). No specific DR antigens or DQB1 alleles were increased in either TBII-negative atrophic autoimmune thyroidities or goitrous autoimmune thyroiditis. A significant decrease in the frequency of HLA-DR6 antigen was observed in both TBII-positive atrophic antoimmune thyroiditis (0% vs. 32%; x[sup 2] = 8.4; Pc = 0.03) and goitrous autoimmune thyroiditis (0% vs. 32%; x[sup 2] = 23.2; Pc < 0.001) patients. The frequency of the HLC-Cwl antigen was significantly increased in all patient groups. The authors conclude that TRBab-positive atrophic autoimmune thyroiditis is immunogenetically different from both goitrous autoimmune thyroiditis and TRBab-negative atrophic autoimmune thyroiditis. It is possible that HLA-DR8 and/or DQB1*0302 may be related to the susceptibility genes involved in the production of TRBab in Koreans. 32 refs., 5 tabs.

  6. Role of salivary anti-SSA/B antibodies for diagnosing primary Sjögren’s syndrome

    PubMed Central

    Wei, Pan; Li, Chunlei; Qiang, Lu; He, Jing; Li, Zhanguo

    2015-01-01

    The diagnosis of primary Sjögren’s syndrome (pSS) is complex, and the saliva test is a potential method to improve the existing diagnostic criteria. Objective: To estimate the diagnostic accuracy of salivary anti-SSA/B antibodies in primary Sjögren’s syndrome (pSS), and to analyze their correlations with clinical and laboratory profiles. Study Design: This study enrolled 100 pSS patients and 140 non-pSS controls, including 40 rheumatoid arthritis (RA) patients, 40 systemic lupus erythematosus (SLE) patients, and 60 healthy controls. Unstimulated whole saliva and stimulated parotid saliva samples were collected from the subjects. Salivary anti-SSA/B antibodies were measured using an enzyme-linked immunosorbent assay (ELISA). Clinical and laboratory data were retrieved from the medical records. Results: In the pSS group, the sensitivity of anti-SSA and anti-SSB antibodies in whole saliva was 49% and 29%, respectively, and the specificity was 87.5% and 95%. The sensitivity of anti-SSA and anti-SSB antibodies in parotid saliva was 32% and 8%, respectively, and the specificity was 95.52% and 97.86%, respectively. In the pSS group, the diagnostic accuracy of anti-SSA/B antibodies in whole saliva was significantly higher than in parotid saliva (p<0.05), but was significantly lower than in serum (p<0.05). The salivary flow rate in the pSS group positive for whole salivary anti-SSA was significantly lower than in the negative group (p<0.05). The prevalence of rheumatoid factor and antinuclear factor were significantly higher in salivary SSB-positive pSS patients than in SSB-negative patients (p<0.05). Conclusions: Compared to parotid saliva, whole saliva is a more suitable diagnostic fluid. Using salivary anti-SSA/B antibodies as a single test item is insufficient given the relatively low sensitivity. Further studies should investigate the possibility of combining tests for different salivary autoantibodies as a method for diagnosing pSS. Key words:Primary Sjögren’s syndrome, salivary diagnostics, anti-SSA autoantibodies, anti-SSB autoantibodies. PMID:25475778

  7. ABO-Incompatible Living Donor Liver Transplantation from Hepatitis B Core Antibody Positive Donor to Hepatitis C Liver Cirrhosis Recipient: A Case Report.

    PubMed

    Umemura, Akira; Nitta, Hiroyuki; Sasaki, Akira; Takahara, Takeshi; Hasegawa, Yasushi; Wakabayashi, Go

    2014-01-01

    Herein, we describe an extremely rare experience of a patient with liver cirrhosis from hepatitis C virus (LC-HCV) who underwent an ABO-incompatible living donor liver transplantation (ABO-I-LDLT) using a hepatitis B core antibody (HBc-Ab) positive donor's liver graft. A 47-year-old Japanese woman with end stage LC-HCV, as a recipient, was preoperatively administered rituximab, mycophenolate mofetil, and steroids without plasma exchange. A routine ABO-I-LDLT procedure was applied using her daughter's HBc-Ab positive liver graft. Prophylaxis of the hepatitis B virus (HBV) infection using hepatitis B immunoglobulin (HBIG) and entecavir had been properly administered. Three months after the ABO-I-LDLT, HCV hepatitis relapsed. To date, this patient has been under antiviral therapy and prophylaxis of HBV infection using HBIG, while entecavir has been continued. The cognitions and techniques with regard to ABO-I-LDLT, prophylaxis of HBV cross infection, various patterns of immunosuppression, and antiviral therapy for HCV relapse are indispensable in managing a transplant recipient. According to the prophylaxis of HBV cross infection under ABO-I-LDLT, it may be very important to keep the HBs-Ab titer higher than usual for HBV naïve recipients, because severe systemic immunosuppression can cause de novo hepatitis. PMID:25045572

  8. Endogenous interleukin 6 production in multiple myeloma patients treated with chimeric monoclonal anti-IL6 antibodies indicates the existence of a positive feed-back loop.

    PubMed Central

    van Zaanen, H C; Koopmans, R P; Aarden, L A; Rensink, H J; Stouthard, J M; Warnaar, S O; Lokhorst, H M; van Oers, M H

    1996-01-01

    In vitro as well as in vivo observations have shown that IL6 plays a key role in the pathogenesis of multiple myeloma. Therefore we started a phase I/II dose escalating study with chimeric monoclonal anti-IL6 antibodies (cMab) in multiple myeloma (MM) patients resistant to second-line chemotherapy. Here we describe the pharmacological data as well as a new method for calculating the endogenous IL6 production. The cMab (CLB IL6/8; Kd: 6.25 x 10(-12) M) was given in two cycles of 14 daily infusions, starting on day 1 and day 28. Daily dose: 5 mg in patients 1-3, 10 mg in patients 4-6, and 20 mg in patients 7-9 (total dose 140, 280, and 560 mg of anti-IL6, respectively). Using the pharmacokinetic data of free IL6 and the binding characteristics of the cMab, the endogenous IL6 production could be calculated from day to day using a one-compartment open model. The median half-life time of this antibody was 17.6 d. No human antichimeric antibodies were induced. Pre-treatment median endogenous IL6 production in the MM patients was 60 micrograms/d (range 13.8-230; normal controls < 7 micrograms/d). During treatment with anti-IL6 cMabs, the endogenous IL6 production immediately decreased in all patients to below 3 micrograms/d and never reached the pre-treatment value during the treatment period, except in two patients who developed an active infection, resulting in an IL6 production of 128 and 1,208 micrograms/d, respectively. We concluded that in MM patients endogenous IL6 production is 2-30 times higher than in healthy individuals. The anti-IL6 cMab strongly suppress this endogenous IL6 production, probably by blocking a positive feed-back loop, but this cMab does not prevent infection-induced IL6 production. The chimeric anti-IL6 Mabs have a long half-life time, a low immunogenicity, and are able to block IL6-dependent processes in vivo. PMID:8823310

  9. Antithyroid microsomal antibody

    MedlinePLUS

    ... to confirm the cause of thyroid problems, including Hashimoto thyroiditis . The test is also used to find ... positive test may be due to: Granulomatous thyroiditis Hashimoto thyroiditis High levels of these antibodies have also ...

  10. Trastuzumab-based treatment of HER2-positive breast cancer: an antibody-dependent cellular cytotoxicity mechanism?

    PubMed Central

    Arnould, L; Gelly, M; Penault-Llorca, F; Benoit, L; Bonnetain, F; Migeon, C; Cabaret, V; Fermeaux, V; Bertheau, P; Garnier, J; Jeannin, J-F; Coudert, B

    2006-01-01

    This study evaluated by immunohistochemistry (IHC) immune cell response during neoadjuvant primary systemic therapy (PST) with trastuzumab in patients with HER2-positive primary breast cancer. In all, 23 patients with IHC 3+ primary breast cancer were treated with trastuzumab plus docetaxel. Pathological complete and partial responses were documented for nine (39%) and 14 (61%) patients, respectively. Case-matched controls comprised patients treated with docetaxel-based PST without trastuzumab (D; n=23) or PST without docetaxel or trastuzumab (non-taxane, non-trastuzumab, NT–NT; n=23). All surgical specimens were blind-analysed by two independent pathologists, with immunohistochemical evaluation of B and T lymphocytes, macrophages, dendritic cells and natural killer (NK) cells. Potential cytolytic cells were stained for Granzyme B and TiA1. HER2 expression was also evaluated in residual tumour cells. Trastuzumab treatment was associated with significantly increased numbers of tumour-associated NK cells and increased lymphocyte expression of Granzyme B and TiA1 compared with controls. This study supports an in vivo role for immune (particularly NK cell) responses in the mechanism of trastuzumab action in breast cancer. These results suggest that trastuzumab plus taxanes lead to enhanced NK cell activity, which may partially account for the synergistic activity of trastuzumab and docetaxel in breast cancer. PMID:16404427

  11. Significant Association between Serum Interleukin-6 and Helicobacter pylori Antibody Levels among H. pylori-Positive Japanese Adults

    PubMed Central

    Nakagawa, Hiroko; Tamura, Takashi; Mitsuda, Yoko; Goto, Yasuyuki; Kamiya, Yoshikazu; Kondo, Takaaki; Wakai, Kenji; Hamajima, Nobuyuki

    2013-01-01

    Background. Interleukin-6 (IL-6) is a multifunctional cytokine produced by many types of cells. Inflammation plays a key role in the pathogenesis of atherosclerosis that is an underlying cause of coronary heart disease (CHD). Since the 1990s, some studies have shown an association between H. pylori infection and CHD, which may be mediated by inflammation. Therefore, this study aimed to evaluate the association between serum anti-H. pylori IgG levels and serum IL-6 levels in H. pylori-infected adults. Methods. We enrolled 158 subjects who visited a clinic located in an urban area to be tested for H. pylori infection, using the 13C-urea breath test, and who were found to be infected and subsequently received eradication. Results. The geometric mean serum IL-6 level was 1.78?pg/mL for men, 1.57?pg/mL for women, and 1.64?pg/mL overall. Logarithms of serum IL-6 levels were positively correlated with logarithms of serum H. pylori IgG levels (r = 0.24, P = 0.002). In multiple linear regression analysis adjusting for sex and age, the serum IL-6 level was still significantly associated with the IgG level in all subjects (? = 0.18, P = 0.012). Conclusion. Higher H. pylori IgG levels were significantly associated with higher serum IL-6 levels among H. pylori-infected individuals. PMID:24453409

  12. Quantitative hepatitis B core antibody level is a new predictor for treatment response in HBeAg-positive chronic hepatitis B patients receiving peginterferon.

    PubMed

    Hou, Feng-Qin; Song, Liu-Wei; Yuan, Quan; Fang, Lin-Lin; Ge, Sheng-Xiang; Zhang, Jun; Sheng, Ji-Fang; Xie, Dong-Ying; Shang, Jia; Wu, Shu-Huan; Sun, Yong-Tao; Wei, Shao-Feng; Wang, Mao-Rong; Wan, Mo-Bin; Jia, Ji-Dong; Luo, Guang-Han; Tang, Hong; Li, Shu-Chen; Niu, Jun-Qi; Zhou, Wei-Dong; Sun, Li; Xia, Ning-Shao; Wang, Gui-Qiang

    2015-01-01

    A recent study revealed that quantitative hepatitis B core antibody (qAnti-HBc) level could serve as a novel marker for predicting treatment response. In the present study, we further investigated the predictive value of qAnti-HBc level in HBeAg-positive patients undergoing PEG-IFN therapy. A total of 140 HBeAg-positive patients who underwent PEG-IFN therapy for 48 weeks and follow-up for 24 weeks were enrolled in this study. Serum samples were taken every 12 weeks post-treatment. The predictive value of the baseline qAnti-HBc level for treatment response was evaluated. Patients were further divided into 2 groups according to the baseline qAnti-HBc level, and the response rate was compared. Additionally, the kinetics of the virological and biochemical parameters were analyzed. Patients who achieved response had a significantly higher baseline qAnti-HBc level (serological response [SR], 4.52±0.36 vs. 4.19±0.58, p=0.001; virological response [VR], 4.53±0.35 vs. 4.22±0.57, p=0.005; combined response [CR], 4.50±0.36 vs. 4.22±0.58, p=0.009)). Baseline qAnti-HBc was the only parameter that was independently correlated with SR (p=0.008), VR (p=0.010) and CR(p=0.019). Patients with baseline qAnti-HBc levels ?30,000 IU/mL had significantly higher response rates, more HBV DNA suppression, and better hepatitis control in PEG-IFN treatment. In conclusion, qAnti-HBc level may be a novel biomarker for predicting treatment response in HBeAg-positive patients receiving PEG-IFN therapy. PMID:25553110

  13. Prevention of de novo hepatitis B with adefovir dipivoxil in recipients of liver grafts from hepatitis B core antibody-positive donors.

    PubMed

    Chang, Matthew S; Olsen, Sonja K; Pichardo, Elsa M; Heese, Scott; Stiles, Jessica B; Abdelmessih, Rita; Verna, Elizabeth C; Guarrera, James V; Emond, Jean C; Brown, Robert S

    2012-07-01

    Lamivudine has been shown to prevent de novo hepatitis B virus (HBV) infections in liver transplantation (LT) patients receiving hepatitis B core antibody-positive (HBcAb(+)) grafts, but it may produce long-term resistance. Adefovir dipivoxil (ADV) might be effective in preventing de novo hepatitis and resistance. A single-center, prospective trial was conducted with 16 adults (10 men and 6 women, mean age = 54 ± 11 years) who underwent LT with HBcAb(+) grafts between September 2007 and October 2009. After LT, patients were given ADV [10 mg daily (adjusted for renal function)]. No hepatitis B immune globulin was administered. At LT, all graft recipients were hepatitis B surface antigen-negative (HBsAg(-)), 38% were surface antibody-positive (HBsAb(+)), and 50% were HBcAb(+). The median follow-up after LT was 1.8 years (range = 1.0-2.6 years). All recipients had undetectable HBV DNA (<40 IU/mL) after LT until the end of follow-up. One recipient (6%) who was HBsAb(-) and HBcAb(-) before LT became HBsAg(+) after 52 weeks. One recipient was switched from ADV to entecavir for chronic renal insufficiency, and 19% of the patients had renal dose adjustments. There was a nonsignificant trend of increasing creatinine levels over time (1.2 mg/dL at LT, 1.3 mg/dL 1 year after LT, and 2.0 mg/dL 2 years after LT, P = 0.27). A comparison with a control cohort of LT recipients with hepatitis C virus who did not receive ADV showed no difference in the creatinine levels at LT or 1 year after LT. In conclusion, ADV prophylaxis prevents HBV replication in recipients of HBcAb(+) livers but does not fully protect recipients from de novo HBV. Long-term follow-up is needed to better determine the risk of de novo infection. PMID:22422699

  14. Anti-nuclear weapons activism in the United States and Great Britain: a comparative analysis

    SciTech Connect

    Sussman, G.

    1987-01-01

    This study is a response to the lacuna in empirical research into political activism and the nuclear issue and seeks to ascertain the social and value characteristics, political attitudes, and political behavior of activists in the United States and Great Britain. Consideration is also given to gender differences in light of evidence of an emerging gender gap in these two countries. The study investigates the common forces cited in two sets of literature - post-industrialism and anti-nuclear weapons movements - which provide a framework for analysis. Survey research data is employed to assess cross-national similarities and differences. The findings obtained indicate that while American and British activists exhibit common social and value characteristics, British activists appear more integrated in their political opposition to nuclear weapons compared with their American counterparts. Survey results indicate that the political-action repertoire of these activists is quite diverse, suggesting a new style of politics in advanced industrial democracies. Gender-based analysis reveals two important findings. First, activist American men differ significantly from the other three social groups in their attitudes towards nuclear weapons. Second, activist women in both national settings participate at a level equal to or exceeding that of activist men.

  15. Antibodies to nucleoprotein and to hydrazide-altered soluble nucleoprotein in tuberculous patients receiving isoniazid

    PubMed Central

    Alarcón-Segovia, D.; Fishbein, Eugenia; Betancourt, V. M.

    1969-01-01

    Antibodies to calf thymus nuclei, nucleoprotein, DNA, soluble nucleoprotein and hydrazide (hydrallazine and isoniazid)-altered nucleoprotein were investigated by a standard complement-fixation method in 214 tuberculous patients receiving isoniazid. Findings were compared to those on thirty-seven sera from lupus patients receiving neither steroids nor immunosuppressants and on sixty-six sera from normal controls. The incidence of antibodies to all antigens studied except DNA was significantly higher in isoniazid-treated tuberculous patients than in the normal controls, but lower than in the lupus patients. Unlike lupus there were no detectable DNA antibodies in the tuberculous or in the control sera. Antibodies to nucleoprotein (soluble and insoluble) and particularly to hydrazide-altered nucleoprotein were the most frequently found in the isoniazid-treated tuberculous patients. In general, antinuclear antibodies were more frequent in the isoniazid-treated tuberculous female than in the male; in the adult than in the child. It is suggested that hydrazides may cause in vivo similar alteration of nucleoprotein to that which they cause in vitro. Hydrazide-altered nucleoprotein probably elicits the production of antinuclear antibodies which in turn may activate systemic lupus erythematosus in otherwise predisposed individuals. PMID:5359961

  16. Amino Acid Changes at Positions 173 and 187 in the Human T-Cell Leukemia Virus Type 1 Surface Glycoprotein Induce Specific Neutralizing Antibodies

    PubMed Central

    Blanchard, Sophie; Astier-Gin, Thérèse; Tallet, Béatrice; Moynet, Daniel; Londos-Gagliardi, Danielle; Guillemain, Bernard

    1999-01-01

    The nucleotide sequence of human T-cell leukemia virus type 1 (HTLV-1) is highly conserved, most strains sharing at least 95% sequence identity. This sequence conservation is also found in the viral env gene, which codes for the two envelope glycoproteins that play a major role in the induction of a protective immune response against the virus. However, recent reports have indicated that some variations in env sequences may induce incomplete cross-reactivity between HTLV-1 strains. To identify the amino acid changes that might be involved in the antigenicity of neutralizable epitopes, we constructed expression vectors coding for the envelope glycoproteins of two HTLV-1 isolates (2060 and 2072) which induced human antibodies with different neutralization patterns. The amino acid sequences of the envelope glycoproteins differed at four positions. Vectors coding for chimeric or point-mutated envelope proteins were derived from 2060 and 2072 HTLV-1 env genes. Syncytium formation induced by the wild-type or mutated envelope proteins was inhibited by human sera with different neutralizing specificities. We thus identified two amino acid changes, I173?V and A187?T, that play an important role in the antigenicity of neutralizable epitopes located in this region of the surface envelope glycoprotein. PMID:10516045

  17. A case of Goodpasture syndrome positive for anti-GBM antibody and MPO-ANCA complicated by a variety of serious infections.

    PubMed

    Sakoda, C; Kusaba, T; Adachi, T; Sonomura, K; Kimura, T; Nakayama, M; Kishimoto, N; Nakagawa, H; Okigaki, M; Hatta, T; Matsubara, H; Mori, Y

    2011-04-01

    A 62-year-old female was admitted to our hospital for investigation of acute progressive renal insufficiency and a systemic inflammatory reaction, despite treatment with several antibiotics. Laboratory data revealed severe renal insufficiency and positive titers for the myeloperoxidase anti-neutrophil cytoplasmic and anti-glomerular basement membrane antibodies. The deterioration of her general status did not allow us to perform the renal biopsy. Although corticosteroid therapy, hemodialysis, and plasma exchange were concomitantly initiated, pulmonary hemorrhage occurred several days after admission. Mechanical ventilation support was provided and continuous hemodiafiltration was carried out, following which the respiratory failure improved immediately. However, she developed clinical depression and suicidal behavior under the intensive therapy. Therefore, plasma exchange was discontinued and corticosteroid was tapered as quickly as possible. Four months after admission, platelet transfusion and short-term mechanical ventilation support improved the pulmonary hemorrhage; however, her mental status deteriorated despite psychiatric consultation and treatment with a tranquilizer. Thereafter, severe and serious systemic infection due to various pathogens including Staphylococcus aureus, Cytomegalovirus, Pneumocystis jiroveci, Pseudomonas aeruginosa, and Bacteroides recurred, and she died from systemic invasive aspergillosis (IA). We suspected severe immunosuppression caused by various factors, such as predonisolone administration, chronic renal failure on maintenance hemodialysis, depression, and malnutrition due to chronic inflammation and granulocytopenia as a side effect of ganciclovir. When treating rapidly progressive glomerulonephritis, immunosuppressive status should be carefully monitored regarding not only the dosage of therapeutic regimen but also the mental health status and nutrition of the patient. PMID:21426895

  18. Detection rate and antigenic specificities of antineutrophil cytoplasmic antibodies in chinese patients with clinically suspected vasculitis.

    PubMed

    Xin, Gang; Zhao, Ming-Hui; Wang, Hai-Yan

    2004-05-01

    The detection rate of antineutrophil cytoplasmic antibodies (ANCA) in Chinese patients with clinically suspected small vessel vasculitis was investigated, and their antigen specificity and demographic features were analyzed. A number of sera (n = 5,604) sent to our referral laboratory for ANCA screening were tested by indirect immunofluorescence (IIF), enzyme-linked immunosorbent assays (ELISAs) for myeloperoxidase (MPO)- and proteinase 3 (PR3)-ANCA. Then the IIF-ANCA-positive sera that were negative for MPO- and PR3-ANCA were further tested by antigen-specific ELISA by using other five highly purified known ANCA antigens as solid-phase ligands. The known antigens included bactericidal/permeability-increasing protein (BPI), human leukocyte elastase (HLE), lactoferrin, cathepsin G, and azurocidins. Of the 5,604 sera, 267 (4.76%) sera were IIF-ANCA positive and 390 (7%) were antinuclear antibody (ANA) positive in the IIF assay. Of the IIF-positive samples, 213 were anti-MPO positive, 32 were anti-PR3 positive, and five cases were positive for both. Of the 48 sera positive for IIF-ANCA but negative for MPO- and PR3-ANCA, 13 sera (27%) recognized other target antigens, 7 sera recognized BPI, 5 recognized HLE, 1 recognize cathepsin G, and 1 recognized azurocidin. None of the sera recognized lactoferrin, and one serum sample recognized both BPI and HLE. The majority of ANCA-positive patients presented in summer or winter. There was no difference in gender (male/female ratio, 1:1.12) in ANCA-positive patients with a mean age of 53.1 years. The male/female ratio was 1.17:1 for patients over 60 years of age; however, it was 1:4 for patients under 20 years of age. We conclude that ANCA-related diseases are not rare in China, and the major antigens are MPO and PR3. When the IIF technique is used to detect ANCA, ANA should be carefully distinguished. PMID:15138182

  19. Chronic Malaria Revealed by a New Fluorescence Pattern on the Antinuclear Autoantibodies Test

    PubMed Central

    Hommel, Benjamin; Charuel, Jean-Luc; Jaureguiberry, Stéphane; Arnaud, Laurent; Courtin, Regis; Kassab, Petra; Prendki, Virginie; Paris, Luc; Ghillani-Dalbin, Pascale; Thellier, Marc; Caumes, Eric; Amoura, Zahir; Mazier, Dominique; Musset, Lucile; Buffet, Pierre; Miyara, Makoto

    2014-01-01

    Background Several clinical forms of malaria such as chronic carriage, gestational malaria or hyper-reactive malarial splenomegaly may follow a cryptic evolution with afebrile chronic fatigue sometimes accompanied by anemia and/or splenomegaly. Conventional parasitological tests are often negative or not performed, and severe complications may occur. Extensive explorations of these conditions often include the search for antinuclear autoantibodies (ANA). Methods We analysed fluorescence patterns in the ANA test in patients with either chronic cryptic or acute symptomatic malaria, then conducted a one-year prospective study at a single hospital on all available sera drawn for ANA detections. We then identified autoantibodies differentially expressed in malaria patients and in controls using human protein microarray. Results We uncovered and defined a new, malaria-related, nucleo-cytoplasmic ANA pattern displaying the specific association of a nuclear speckled pattern with diffuse cytoplasmic perinuclearly-enhanced fluorescence. In the one-year prospective analysis, 79% of sera displaying this new nucleo-cytoplasmic fluorescence were from patients with malaria. This specific pattern, not seen in other parasitic diseases, allowed a timely reorientation of the diagnosis toward malaria. To assess if the autoantibody immune response was due to autoreactivity or molecular mimicry we isolated 42 autoantigens, targets of malarial autoantibodies. BLAST analysis indicated that 23 of recognized autoantigens were homologous to plasmodial proteins suggesting autoimmune responses directly driven by the plasmodial infection. Conclusion In patients with malaria in whom parasitological tests have not been performed recognition of this new, malaria-related fluorescence pattern on the ANA test is highly suggestive of the diagnosis and triggers immediate, easy confirmation and adapted therapy. PMID:24551116

  20. Degradation of a monoclonal anti-mu chain antibody in a human surface IgM-positive B cell line starts in prelysosomal vesicle.

    PubMed

    Ruud, E; Kindberg, G M; Blomhoff, H K; Godal, T; Berg, T

    1988-11-01

    The surface IgM-mediated endocytosis and intracellular transport of an anti-F(c mu)5 mAb was studied by using subcellular fractionation in sucrose gradients. The results of such experiments showed that antibody was initially endocytosed in vesicles of low density, and later transferred to a presumably lysosomal compartment of higher density. SDS-PAGE analysis of gradient fractions showed that high Mr degradation fragments of the endocytosed antibody were formed in the low density vesicles before terminal degradation could be recorded. The partial degradation of the antibody was not blocked by low temperature or enzyme inhibitors, such as leupeptin and benzyloxycarbonyl-phenylalanylalanine-diazomyethyl-ketone, all of which severely retarded terminal degradation. The data also suggested that the recycling of partially degraded antibody to the cell surface employed a pool of such low density prelysosomal vesicles. PMID:3139770

  1. Chicken egg yolk antibodies against F18ab fimbriae of Escherichia coli inhibit shedding of F18 positive E. coli by experimentally infected pigs

    Microsoft Academic Search

    H. Imberechts; P. Deprez; E. Van Driessche; P. Pohl

    1997-01-01

    F18ab and F18ac are antigenic variants of a colonizing fimbria commonly found on E. coli associated with postweaning diarrhea and edema disease in pigs. Chicken F18ab antibodies were obtained by immunising hens with purified F18ab fimbriae. For their in vitro characterisation antibodies were isolated from diluted egg yolks by ammonium sulfate precipitation. In vitro adhesion tests demonstrated that the chicken

  2. Anti-oxidized low-density lipoprotein antibodies in myeloperoxidase–positive vasculitis patients preferentially recognize hypochlorite-modified low density lipoproteins

    PubMed Central

    Slot, M C; Theunissen, R; van Paassen, P; Damoiseaux, J G M C; Cohen Tervaert, J W

    2007-01-01

    Many patients surviving vasculitis are prone to accelerated atherosclerosis and often have enhanced levels of antibodies to oxidized low-density lipoprotein (oxLDL). To measure anti-oxLDL antibodies, oxidation of LDL is achieved with copper (Cu) or malondialdehyde (MDA). Because, in vivo, LDL may be oxidized with myeloperoxidase (MPO) or its product hypochlorite, we measured anti-hypochlorite LDL antibodies in patients with vasculitis, haemodialysis patients and healthy controls. A newly developed enzyme-linked immunosorbent assay (ELISA) was used to detect antibodies to oxLDL as modified by hypochlorite. Results are compared with data obtained by standard LDL oxidation using MDA–LDL or Cu–LDL as substrate. Results were compared between anti-neutrophil cytoplasmic antibodies (ANCA)-associated vasculitis (AAV) patients (n = 93), haemodialysis (HD) patients (n = 59) and healthy controls (HC; n = 43). Furthermore, patients with MPO–ANCA-associated vasculitis (n = 47) were compared with patients with proteinase 3 (PR3)–ANCA associated vasculitis (n = 46). Optimal cut-off points were determined by receiver operator characteristic (ROC) curve analysis. Anti-oxLDL antibodies are enhanced in AAV patients (MDA–LDL and hypochlorite–LDL) and in HD patients (hypochlorite–LDL), when compared to HC. Furthermore, patients with MPO–ANCA-associated vasculitis had higher levels of antibodies to hypochlorite–LDL than patients with PR3–ANCA-associated vasculitis. Our newly developed assay, in which hypochlorite–LDL is used as substrate, seems a more sensitive assay than traditional assays to measure oxLDL antibodies. Furthermore, our results suggest that enhanced MPO-mediated LDL oxidation occurs in patients with MPO–ANCA. PMID:17521320

  3. Antiphospholipid antibodies during 6-month treatment with infliximab: A preliminary report

    PubMed Central

    Kolarz, Bogdan; Majdan, Maria; Darmochwa?-Kolarz, Dorota A.; Dryglewska, Magdalena

    2014-01-01

    Background The introduction of tumor necrosis factor (TNF) antagonists (adalimumab, infliximab, and etanercept) was a major advance and was highly important and beneficial in most rheumatoid arthritis (RA) patients. The adverse effects of this treatment are infrequent, but include opportunistic intracellular infection (especially the reactivation of latent Mycobacterium tuberculosis); exacerbation of demyelinating disorders; and the production of various types of antibodies such as antinuclear antibodies (ANA) or double-stranded DNA autoantibodies (dsDNA) and antiphospholipid antibodies (aPL) such as anti-cardiolipin antibodies (aCL) and anti-B2GP-I antibodies (B2GP-I). The aim of the study was to determine the prevalence of aCL and B2GP-I in IgM and IgG classes, using ELISA tests, during 6 months of follow-up in patients with refractory RA successfully treated with infliximab. Material/Methods We determined the prevalence of aCL and B2GP-I in IgM and IgG classes, using ELISA tests, during 6 months of follow-up in patients with refractory RA successfully treated with infliximab. Results We observed a statistically important increase only in the group of B2GP-I IgM (p<0.05). There are contradictory results concerning the ability of infliximab to induce aPL, but most authors confirm this phenomenon. Conclusions Further investigations are needed to determine if the new aPL appears in patients with ?2-GPI gene polymorphisms such as leucine-to-valine substitution at position 247, which can lead to a conformational changes in ?2-GPI protein, leading to aPL synthesis. The role of aPL in pathogenesis of APS is still unclear, but we should remember the immunogenic aspect of TNF antagonist treatment. Therefore, we recommend early detection of aPL and observation of the patient, paying special attention to signs and symptoms of thromboembolism. PMID:25027437

  4. Human Germline Antibody Gene Segments Encode Polyspecific Antibodies

    PubMed Central

    Willis, Jordan R.; Briney, Bryan S.; DeLuca, Samuel L.; Crowe, James E.; Meiler, Jens

    2013-01-01

    Structural flexibility in germline gene-encoded antibodies allows promiscuous binding to diverse antigens. The binding affinity and specificity for a particular epitope typically increase as antibody genes acquire somatic mutations in antigen-stimulated B cells. In this work, we investigated whether germline gene-encoded antibodies are optimal for polyspecificity by determining the basis for recognition of diverse antigens by antibodies encoded by three VH gene segments. Panels of somatically mutated antibodies encoded by a common VH gene, but each binding to a different antigen, were computationally redesigned to predict antibodies that could engage multiple antigens at once. The Rosetta multi-state design process predicted antibody sequences for the entire heavy chain variable region, including framework, CDR1, and CDR2 mutations. The predicted sequences matched the germline gene sequences to a remarkable degree, revealing by computational design the residues that are predicted to enable polyspecificity, i.e., binding of many unrelated antigens with a common sequence. The process thereby reverses antibody maturation in silico. In contrast, when designing antibodies to bind a single antigen, a sequence similar to that of the mature antibody sequence was returned, mimicking natural antibody maturation in silico. We demonstrated that the Rosetta computational design algorithm captures important aspects of antibody/antigen recognition. While the hypervariable region CDR3 often mediates much of the specificity of mature antibodies, we identified key positions in the VH gene encoding CDR1, CDR2, and the immunoglobulin framework that are critical contributors for polyspecificity in germline antibodies. Computational design of antibodies capable of binding multiple antigens may allow the rational design of antibodies that retain polyspecificity for diverse epitope binding. PMID:23637590

  5. Antibody Purification AntibodyPurification

    E-print Network

    Lebendiker, Mario

    Antibody Purification AntibodyPurification 24 Overview Antibodies are proteins; therefore, methods forms of general protein purification methods (see the Protein Purification section of the PierceG. Protein L is a third protein of bacterial origin that has been developed for use in affinity purification

  6. Antineutrophil cytoplasmic antibody (ANCA)-associated autoimmune diseases induced by antithyroid drugs: comparison with idiopathic ANCA vasculitides.

    PubMed

    Bonaci-Nikolic, Branka; Nikolic, Milos M; Andrejevic, Sladjana; Zoric, Svetlana; Bukilica, Mirjana

    2005-01-01

    Clinical and serological profiles of idiopathic and drug-induced autoimmune diseases can be very similar. We compared data from idiopathic and antithyroid drug (ATD)-induced antineutrophil cytoplasmic antibody (ANCA)-positive patients. From 1993 to 2003, 2474 patients were tested for ANCA in the Laboratory for Allergy and Clinical Immunology in Belgrade. Out of 2474 patients, 72 (2.9%) were anti-proteinase 3 (PR3)- or anti-myeloperoxidase (MPO)-positive and their clinical and serological data were analyzed. The first group consisted of ANCA-associated idiopathic systemic vasculitis (ISV) diagnosed in 56/72 patients: 29 Wegener's granulomatosis (WG), 23 microscopic polyangiitis (MPA) and four Churg-Strauss syndrome. The second group consisted of 16/72 patients who became ANCA-positive during ATD therapy (12 receiving propylthiouracil and four receiving methimazole). We determined ANCA and antinuclear (ANA) antibodies by indirect immunofluorescence; PR3-ANCA, MPO-ANCA, anticardiolipin (aCL) and antihistone antibodies (AHA) by ELISA; and cryoglobulins by precipitation. Complement components C3 and C4, alpha-1 antitrypsin (alpha1 AT) and C reactive protein (CR-P) were measured by nephelometry. Renal lesions were present in 3/16 (18.8%) ATD-treated patients and in 42/56 (75%) ISV patients (p <0.001). Skin lesions occurred in 10/16 (62.5%) ATD-treated patients and 14/56 (25%) ISV patients (p <0.01). ATD-treated patients more frequently had MPO-ANCA, ANA, AHA, aCL, cryoglobulins and low C4 (p <0.01). ISV patients more frequently had low alpha1 AT (p = 0.059) and high CR-P (p <0.001). Of 16 ATD-treated patients, four had drug-induced ANCA vasculitis (three MPA and one WG), while 12 had lupus-like disease (LLD). Of 56 ISV patients, 13 died and eight developed terminal renal failure (TRF). There was no lethality in the ATD-treated group, but 1/16 with methimazole-induced MPA developed pulmonary-renal syndrome with progression to TRF. ANCA-positive ISV had a more severe course in comparison with ATD-induced ANCA-positive diseases. Clinically and serologically ANCA-positive ATD-treated patients can be divided into two groups: the first consisting of patients with drug-induced WG or MPA which resemble ISV and the second consisting of patients with LLD. Different serological profiles could help in the differential diagnosis and adequate therapeutic approach to ANCA-positive ATD-treated patients with symptoms of systemic disease. PMID:16207324

  7. Passive protection of suckling infant mice against F41-positive enterotoxigenic Escherichia coli strains by intravenous inoculation of the dams with monoclonal antibodies against F41.

    PubMed

    Duchet-Suchaux, M; Menanteau, P; van Zijderveld, F G

    1992-07-01

    Ten monoclonal antibodies (MAbs) against five different epitope clusters of adhesion factor F41 (two MAbs per cluster) were tested for protection of infant mice against an oral challenge with F41-positive enterotoxigenic Escherichia coli (ETEC) B2C and B41M. Infant mice suckling dams intravenously inoculated with MAbs were orally challenged, and the survival rates were measured for 12 days after inoculation and challenge. Irrespective of their epitope specificity, all F41 MAbs given in a single dose of 4 mg per dam had a protective effect against both ETEC strains. In contrast, one K99 MAb of the same isotype and given in the same dose as the F41 MAbs did not protect infant mice at all. A reduction in the dose of F41 MAbs to 0.032 mg per dam resulted in a decrease in protection. Two different MAbs against the same epitope cluster were not necessarily equally protective. Combining MAbs two by two, whether the MAbs recognized the same epitope cluster or not, resulted in protective activity essentially similar to that obtained with each MAb separately, without any improvement. Therefore, one MAb against any epitope may be sufficient for protection. Enzyme-linked immunosorbent assay (ELISA) titers of MAbs in the serum of dams were similar, irrespective of the epitope specificity of the MAbs, and gradually decreased from day 1 to day 12 after inoculation. We found a good correlation between colostrum and milk ELISA titers of MAbs and serum ELISA titers of MAbs. Colostrum and milk MAb titers were 10-fold lower than corresponding serum MAb titers and stayed high until day 5 after inoculation. The most protective MAb had the highest ELISA titers in colostrum and milk for the first 5 days after inoculation. ETEC strain B2C colonized the intestines of infant mice suckling MAb-inoculated mothers until day 12 after challenge. Intestinal levels of the challenge strain were high on day 2 but never reached the very high numbers (10(9) to 10(10)) described previously in a diarrheic infant mouse model. MAbs did not eliminate the challenge ETEC strain from the intestines of infant mice. PMID:1351882

  8. DNA mobility shift assay as a tool for the detection of anti-dsDNA antibodies in sera from discoid lupus erythematosus patients.

    PubMed

    Keyhani, Jacqueline; Ahadi, Mashallah; Keyhani, Ezzatollah; Naraghi, Zahra; Shamohammadi, Safar

    2012-07-01

    Discoid lupus erythematosus (DLE) is a form of local inflammatory autoimmune disease limited to the skin, involving essentially the face, scalp and ear. DLE occurs in genetically predisposed individuals, sunlight being an identified trigger. Diagnosis is made by clinical examination and histopathology; laboratory tests occasionally performed include anti-nuclear antibodies titers and presence of circulating antibodies against dsDNA. DLE patients have about a 10% chance of developing systemic lupus erythematosus (SLE), a systemic inflammatory autoimmune disease affecting a range of internal organs. Although elevated titers of anti-dsDNA antibodies is an earmark for lupus disease, they are detected in only 20-55% of DLE patients by routine laboratory tests such as enzyme-linked immunosorbent assay (ELISA). In this research, we applied an electrophoretic mobility shift assay (EMSA) in parallel with an ELISA for the detection of circulating anti-dsDNA in DLE patients. The assays were conducted on sera as well as on the immunoglobulin G fraction from sera of 24 DLE patients and of 24 healthy individuals. The EMSA was positive for all DLE patients while the ELISA was positive for only 36% of them; both assays were negative for the healthy individuals. EMSA conducted on the sera of 15 patients with lichen planus was negative in all cases. Results suggest that the EMSA is more sensitive than the routine tests used for the detection of anti-dsDNA in DLE, thus helping to improve early detection of the disease and, by extension, to better evaluate the factors triggering the disease. PMID:22486312

  9. Lupus nephritis associated with positive MPO-ANCA in a patient with underlying autoimmune hemolytic anemia

    Microsoft Academic Search

    Yuki Hirai; Masayuki Iyoda; Takanori Shibata; Eijin Ashikaga; Nozomu Hosaka; Hiroki Suzuki; Hisako Nagai; Masanori Mukai; Hirokazu Honda; Aki Kuroki; Kozo Kitazawa; Tadao Akizawa

    2008-01-01

    A 19-year-old female was admitted with general malaise and systemic edema. She had been diagnosed as having autoimmune hemolytic\\u000a anemia (AIHA) eight years earlier and was successfully managed with oral prednisolone. During the current admission, she was\\u000a diagnosed as having systemic lupus erythematosus (SLE) based on the presence of renal involvement, hematological abnormalities,\\u000a and antinuclear and anti-double-stranded DNA antibodies, along

  10. [Anti-PCNA antibody].

    PubMed

    Yamanaka, K; Takasaki, Y

    1993-08-01

    Antibodies to the proliferating cell nuclear antigen (PCNA) detected in patients with systemic lupus erythematosus (SLE) were allowed to react with a nuclear antigen expressed predominantly in proliferating cells such as cultured cells and mitogen-transformed cells. The characterization of both structure and function of PCNA has been studied. PCNA has been identified as a protein with a molecular weight of about 33 kD and isoelectric point of 4.8. The expression of PCNA increased in the cell from the late G1 and S phases of the cell cycle immediately preceding DNA synthesis. Recent studies have revealed that the auxiliary protein of DNA polymerase-d is identical to PCNA. Anti-PCNA antibodies can be detected by the methods of immunofluorescence (IF), double immunodiffusion (DID), counterimmunoelectrophoresis (CIE), enzyme-linked immunosorbent assay (ELISA) and immunoblotting (IB). Anti-PCNA antibodies were specifically detected in 2-5% of the patients with SLE by DID. In spite of their low frequency, anti-PCNA antibodies are useful as a clinical marker for SLE. Several reports indicated that patients with PCNA positive SLE showed a high frequency of renal and central nerve system (CNS) involvements and thrombocytopenia. In these patients, the anti-PCNA antibody titer was elevated before the development by proteinuria and the titer of anti-PCNA antibodies was decreased by treatment with corticosteroids. In addition, anti-PCNA antibodies have been known as a useful tool to detect activated lymphocytes and malignant proliferating cell. PMID:8103808

  11. Prolonged infections associated with antineutrophil cytoplasmic antibodies specific to proteinase 3 and myeloperoxidase: diagnostic and therapeutic challenge.

    PubMed

    Bonaci-Nikolic, Branka; Andrejevic, Sladjana; Pavlovic, Milorad; Dimcic, Zoran; Ivanovic, Branislava; Nikolic, Milos

    2010-08-01

    Chronic infections may mimic antineutrophil cytoplasmic antibody (ANCA)-associated vasculitides (AAV). We investigated which markers may help in the diagnosis and the prognosis of infections associated with proteinase 3 (PR3) and myeloperoxidase (MPO)-ANCA. In this study (1993-2008)--with an average follow-up of 5.1 years--we compared 66 AAV patients with 17 PR3 and/or MPO-ANCA-positive patients with protracted bacterial (11/17) or viral (6/17) infections. Seven of 17 patients had subacute bacterial endocarditis (SBE), while six of 17 patients had various autoimmune manifestations of chronic hepatitis C virus (HCV) infection. We determined ANCA, antinuclear antibodies, anti-PR3, anti-MPO, anticardiolipin (aCL), antibeta 2 glycoprotein I (beta2-GP I), cryoglobulins, C3, and C4. Patients with infections were younger than AAV patients (p < 0.01). There was no difference in frequency of renal and skin lesions. AAV patients more frequently had pulmonary and nervous system manifestations (p < 0.01). Patients with infections more frequently had dual ANCA (high PR3, low MPO), aCL, anti-beta2-GP I, cryoglobulins, and hypocomplementemia (p < 0.001). Immunosuppressive therapy (IST) was used in five 17 patients who had persistently high ANCA, cryoglobulinemia, and hypocomplementemia. There was no difference in frequency of lethality and renal failure in the two study groups. In patients who are PR3- and/or MPO-ANCA positive, SBE and HCV infection should be excluded. Although similar in renal and skin manifestations in comparison to AAV, only patients with infections developed multiple serological abnormalities. In patients with infections, concomitant presence of ANCA, cryoglobulins, and hypocomplementemia was associated with severe glomerulonephritis. The serological profile should be repeated after specific antimicrobial or surgical therapy, since some cases might require IST. PMID:20306213

  12. Investigation of the prevalence and clinical associations of antibodies to human fibronectin in systemic lupus erythematosus.

    PubMed Central

    Atta, M S; Lim, K L; Ala'deen, D A; Powell, R J; Todd, I

    1995-01-01

    OBJECTIVES--To assess the prevalence of antibodies to human fibronectin (anti-Fn) in sera of patients with certain connective tissue diseases and to determine their association with disease activity and the pattern of organ involvement in patients with systemic lupus erythematosus (SLE). METHODS--A capture enzyme linked immunosorbent assay (ELISA) was developed to quantify anti-Fn antibodies in serum samples from 65 patients with well characterised SLE, 50 with rheumatoid arthritis (RA), 15 with Behçet's disease (BD), 15 with systemic vasculitis and 36 healthy subjects. An anti-Fn antibody titre greater than mean + 3SD of the healthy control log values after back transformation to the normal scale was considered positive. Disease activity in SLE patients was scored using the British Isles Lupus Assessment Group (BILAG) Index. Erythrocyte sedimentation rate (ESR), concentrations of anti-dsDNA antibody, soluble interleukin-2 receptors (sIL-2R), C3, C4, C3 degradation products (C3dg) and immunoglobulin, and antinuclear antibody (ANA) titres were measured in blood samples from SLE patients; neopterin concentration was measured in corresponding urine samples. RESULTS--Anti-Fn antibodies were found in 22 of 65 SLE patients (33.8%), seven of 50 with RA (14%), one of 15 with BD (6.6%) and none of the 15 subjects with vasculitis. Thirty SLE patients had active disease and 35 had inactive disease; their median anti-Fn concentrations were 117 u/ml (range 47-450) and 68 u/ml (range 17-334), respectively (p = 0.0001). The presence of anti-Fn did not correlate with immunoglobulin concentrations or ANA titres in these sera. No significant difference was found between SLE patients with disease activity in one major organ system compared with multiple organ involvement, as defined by BILAG (p = 0.19). However, patients with musculoskeletal manifestations had consistently greater anti-Fn concentrations compared with patients with other clinical manifestations. There were significant correlations between amounts of anti-Fn in SLE sera and ESR (rs = 0.25, p = 0.045), sIL-2R (rs = 0.28, p = 0.024) and urine neopterin (rs = 0.3, p = 0.016) but not with serum anti-dsDNA antibody titres, plasma C3, C3dg or C4. However multiple regression analysis showed a low significant correlation only with sIL-2R and BILAG score (p = 0.047 and 0.042, respectively). CONCLUSION--Anti-Fn antibodies were detected in 34% of SLE patients and in small proportions of RA and BD patients. An association between serum anti-Fn and disease activity in SLE has been identified and most SLE patients with musculoskeletal involvement had increased anti-Fn antibody concentrations. Images PMID:7702398

  13. RhD Specific Antibodies Are Not Detectable in HLA-DRB1*1501 Mice Challenged with Human RhD Positive Erythrocytes

    PubMed Central

    Bernardo, Lidice; Denomme, Gregory A.; Shah, Kunjlata; Lazarus, Alan H.

    2014-01-01

    The ability to study the immune response to the RhD antigen in the prevention of hemolytic disease of the fetus and newborn has been hampered by the lack of a mouse model of RhD immunization. However, the ability of transgenic mice expressing human HLA DRB1*1501 to respond to immunization with purified RhD has allowed this question to be revisited. In this work we aimed at inducing anti-RhD antibodies by administering human RhD+ RBCs to mice transgenic for the human HLA DRB1*1501 as well as to several standard inbred and outbred laboratory strains including C57BL/6, DBA1/J, CFW(SW), CD1(ICR), and NSA(CF-1). DRB1*1501 mice were additionally immunized with putative extracellular immunogenic RhD peptides. DRB1*1501 mice immunized with RhD+ erythrocytes developed an erythrocyte-reactive antibody response. Antibodies specific for RhD could not however be detected by flow cytometry. Despite this, DRB1*1501 mice were capable of recognizing immunogenic sequences of Rh as injection with Rh peptides induced antibodies reactive with RhD sequences, consistent with the presence of B cell repertoires capable of recognizing RhD. We conclude that while HLA DRB1*1501 transgenic mice may have the capability of responding to immunogenic sequences within RhD, an immune response to human RBC expressing RhD is not directly observed. PMID:25628657

  14. RhD Specific Antibodies Are Not Detectable in HLA-DRB1(*)1501 Mice Challenged with Human RhD Positive Erythrocytes.

    PubMed

    Bernardo, Lidice; Denomme, Gregory A; Shah, Kunjlata; Lazarus, Alan H

    2014-01-01

    The ability to study the immune response to the RhD antigen in the prevention of hemolytic disease of the fetus and newborn has been hampered by the lack of a mouse model of RhD immunization. However, the ability of transgenic mice expressing human HLA DRB1(*)1501 to respond to immunization with purified RhD has allowed this question to be revisited. In this work we aimed at inducing anti-RhD antibodies by administering human RhD(+) RBCs to mice transgenic for the human HLA DRB1(*)1501 as well as to several standard inbred and outbred laboratory strains including C57BL/6, DBA1/J, CFW(SW), CD1(ICR), and NSA(CF-1). DRB1(*)1501 mice were additionally immunized with putative extracellular immunogenic RhD peptides. DRB1(*)1501 mice immunized with RhD(+) erythrocytes developed an erythrocyte-reactive antibody response. Antibodies specific for RhD could not however be detected by flow cytometry. Despite this, DRB1(*)1501 mice were capable of recognizing immunogenic sequences of Rh as injection with Rh peptides induced antibodies reactive with RhD sequences, consistent with the presence of B cell repertoires capable of recognizing RhD. We conclude that while HLA DRB1(*)1501 transgenic mice may have the capability of responding to immunogenic sequences within RhD, an immune response to human RBC expressing RhD is not directly observed. PMID:25628657

  15. Anti-idiotypic antibody against anti-DNA in sera of laboratory personnel exposed to lupus sera or nucleic acids.

    PubMed Central

    Hatfield, M; Evans, M; Suenaga, R; Hassanein, K M; Abdou, N I

    1987-01-01

    We tested for anti-DNA, anti-idiotypic, antinuclear, and lymphocytotoxic antibodies in the sera of three groups of normals: volunteers never exposed to lupus sera or nucleic acids (group I), research personnel handling nucleic acids (group II), and laboratory personnel handling lupus sera (group III). There was no significant differences among the groups with respect to levels of either single stranded or double stranded anti-DNA. Group I showed no significant differences in binding to F(ab')2 fragments of lupus anti-DNA, lupus non-anti-DNA or normal IgG. Compared to group I, groups II and III bound significantly higher to anti-DNA F(ab')2 fragments compared to non-anti-DNA F(ab')2 or normal F(ab')2 fragments. Sera from the three groups were negative for antibodies and all but one individual from group III had normal antinuclear antibody titres. These results indicate that sera of normals exposed to lupus sera or to nucleic acids contain an anti-idiotype directed against anti-DNA antibody. The possible role of these anti-idiotypes in regulating the anti-DNA antibody is discussed. PMID:3500815

  16. The risk of hepatitis B virus reactivation and the role of antiviral prophylaxis in hepatitis B surface antigen negative/hepatitis B core antibody positive patients with diffuse large B-cell lymphoma receiving rituximab-based chemotherapy.

    PubMed

    Lu, Shasha; Xu, Yu; Mu, Qitian; Cao, Lihong; Chen, Jian; Zhu, Zhijuan; Lou, Yinjun; Meng, Haitao; Qian, Wenbin; Tong, Hongyan; Mai, Wenyuan; Huang, Jian; Yu, Wenjuan; Zhao, Xiaoying; Jin, Jie

    2015-04-01

    The risk factors and the role of prophylactic antiviral therapy of hepatitis B virus (HBV) reactivation in patients with hepatitis B surface antigen (HBsAg) negative/hepatitis B core antibody (HBcAb) positive disease remain controversial. We reviewed 629 patients with diffuse large B-cell lymphoma (DLBCL). Among 629 patients, 150 of 246 patients with resolved HBV (HBsAg negative and HBcAb positive) were treated with rituximab-combined therapy. Among these 150 patients, none of 104 patients (0.0%) who were hepatitis B surface antibody (HBsAb) positive experienced HBV reactivation versus four of 46 patients (8.7%) who were HBsAb negative (p = 0.008). One of 113 patients (0.9%) with International Prognostic Index (IPI) 0-2 suffered HBV reactivation versus three of the remaining 37 patients (8.1%) with IPI 3-5 (p = 0.047). HBsAb and IPI are potential risk factors for HBV reactivation. The use of prophylactic agents may not be recommended for these patients until the occurrence of HBV reactivation. PMID:25065698

  17. The Clinical and Genomic Significance of Donor-Specific Antibody–Positive/C4d-Negative and Donor-Specific Antibody–Negative/C4d-Negative Transplant Glomerulopathy

    PubMed Central

    Hayde, Nicole; Bao, Yi; Pullman, James; Ye, Bin; Calder, R. Brent; Chung, Monica; Schwartz, Daniel; Lubetzky, Michelle; Ajaimy, Maria; de Boccardo, Graciela

    2013-01-01

    Summary Background This study investigated the mechanisms involved in development of donor-specific antibody (DSA) and/or C4d-negative transplant glomerulopathy (TGP) by allograft gene expression profiles using microarrays. Design, Setting, Participants, & Measurements This cohort study was conducted in kidney transplant recipients. Patients were eligible for inclusion if they required a clinically indicated biopsy at any time point after their transplant. They were then classified according to their histopathology findings and DSA and C4d results. Eighteen chronic antibody-mediated rejection (CAMR), 14 DSA+/C4d? TGP, 25 DSA?/C4d? TGP, and 47 nonspecific interstitial fibrosis/tubular atrophy (IFTA) biopsy specimens were identified. In a subset of patients from the study population, biopsy specimens in each group and normal transplant kidney specimens were analyzed with Affymetrix Human Gene 1.0 ST Arrays. Results The mean sum score of glomerulitis and peritubular capillaritis increased from 0.28±0.78 in IFTA specimens to 0.75±0.85 in DSA?/C4d? TGP specimens, 1.71±1.49 in DSA+/C4d?/TGP specimens, and 2.11±1.74 in CAMR specimens (P<0.001). During a median follow-up time of 2 (interquartile range, 1.4–2.8) years after biopsy, graft loss was highest in CAMR specimens (27.8%) compared to IFTA specimens (8.5%), DSA+/C4d? TGP specimens (14.3%), and DSA?/C4d? TGP specimens (16%) (P=0.01). With use of microarrays, comparison of the gene expression profiles of DSA?/C4d? TGP specimens with glomerulitis + peritubular capillaritis scores > 0 to normal and IFTA biopsy specimens revealed higher expression of quantitative cytotoxic T cell–associated transcripts (QCAT). However, both CAMR and DSA+/C4d? TGP specimens had higher expression of not only QCAT but also IFN-? and rejection-induced, constitutive macrophage-associated, natural killer cell–associated, and DSA-selective transcripts. Endothelial cell–associated transcript expression was upregulated only in CAMR biopsy specimens. Conclusions These results suggested that DSA+/C4d? TGP biopsy specimens may be classified as CAMR. In contrast, DSA?/C4d? TGP specimens showed increased cytotoxic T cell–associated transcripts, suggesting T cell activation as a mechanism of injury. PMID:24030736

  18. Thyroid Antibodies

    MedlinePLUS

    ... autoimmune disease . A low level of thyroid hormones ( hypothyroidism ) can cause symptoms, such as: Weight gain Fatigue ... drug therapy that has associated risks of developing hypothyroidism when thyroid peroxidase antibodies are present, such as ...

  19. Clinical and Therapeutic Aspects Associated to Phospholipid Binding Antibodies (Lupus Anticoagulant and Anticardiolipin Antibodies)

    Microsoft Academic Search

    J. Ordi-Ros; P. Pérez-Pemán; J. Monasterio

    1994-01-01

    Antiphospholipid antibodies (APA) comprise a family of immunoglobulins characterized by their pattern of reactivity in a number of laboratory tests. Included in this family are lupus anticoagulant (LA) anticardiolipin antibodies (ACA) and antibodies causing biologic false positive serologic tests for syphilis (BFP-STS). LA and ACA occur in a variety of conditions, including other autoimmunes disorders, infectious diseases, neoplasic disorders, in

  20. [Antineutrophil cytoplasmic antibodies and associated diseases].

    PubMed

    Sghiri, R; Meddeb, H; Bouguila, J; Jarray, M; Bahri, F; Nouira, R; Zellama, D; Achour, A; Essoussi, A S; Harbi, A; Ghedira, I

    2009-07-01

    Antineutrophil cytoplasmic antibodies are classical serological markers of small-vessels vasculitis. However, they have been described in many other pathological situations. The aim of this study was to determine through our experience, the main antineutrophil cytoplasmic antibodies-associated diseases and to investigate antigen targets of these antibodies. Forty complete observations of antineutrophil cytoplasmic antibodies (ANCA) positive patients either by indirect immunofluorescence or by enzyme immunoassay were analysed. Only five (12.5%) patients have small-vessels vasculitis. Among these, antineutrophil cytoplasmic antibodies were detected only by Elisa in one patient and they were exclusively directed against bactericidal permeability increasing protein in another one. Our study confirms the presence of antineutrophil cytoplasmic antibodies in different diseases. It demonstrates that antineutrophil cytoplasmic antibodies should be investigated by Elisa when indirect immunofluorescence is negative. In small-vessels vasculitis, Proteinase 3 and myeloperoxidase are mainly but not exclusively the antigenic targets of antineutrophil cytoplasmic antibodies. PMID:18834675

  1. Human lymphocyte subpopulations. Human thymus-lymphoid tissue (HTL) antigen-positive lymphocytes forming rosettes with sheep erythrocytes and HTL antigen-negative lymphocytes interacting with antigen-antibody-complement complexes

    PubMed Central

    Yata, J.; Tsukimoto, I.; Tachibana, T.

    1973-01-01

    Human lymphocytes from various lymphoid tissues were studied for the relationship between the existence of HTL (human thymus-lymphoid tissue) antigen, and binding of sheep erythrocytes (E) or sheep erythrocyte–antibody-complement complexes (EA(IgM)C43). E adhered to the majority of thymus lymphocytes and formed rosettes. These lymphocytes were shown to be HTL antigen positive by immunofluorescence performed simultaneously. In the peripheral lymphoid tissues, 10–30% of lymphocytes formed E rosettes and almost all E rosette-forming lymphocytes were HTL antigen positive. Conversely HTL antigen-negative cells did not form E rosettes. In contrast, the cells binding EA(IgM)C43 were always HTL antigen negative. There were very few HTL antigen-positive or rosette-forming lymphocytes either with E or EA(IgM)C43 in bone marrow. From these data we conclude that E-rosette-forming and HTL antigen-positive lymphocytes are of thymus origin and EA(IgM)C43-rosette-forming cells are not thymus-dependent cells. ImagesFig. 1 PMID:4579778

  2. Bispecific antibodies.

    PubMed

    Kontermann, Roland E; Brinkmann, Ulrich

    2015-07-01

    Bispecific antibodies (bsAbs) combine specificities of two antibodies and simultaneously address different antigens or epitopes. BsAbs with 'two-target' functionality can interfere with multiple surface receptors or ligands associated, for example with cancer, proliferation or inflammatory processes. BsAbs can also place targets into close proximity, either to support protein complex formation on one cell, or to trigger contacts between cells. Examples of 'forced-connection' functionalities are bsAbs that support protein complexation in the clotting cascade, or tumor-targeted immune cell recruiters and/or activators. Following years of research and development (R&D), the first bsAb was approved in 2009. Another bsAb entered the market in December 2014 and several more are in clinical trials. Here, we describe the potentials of bsAbs to become the next wave of antibody-based therapies, focusing on molecules in clinical development. PMID:25728220

  3. The pro-solar, anti-nuclear movement: A philosophical perspective

    NASA Astrophysics Data System (ADS)

    Adler, Carl G.

    1983-06-01

    An analysis is presented of the nontechnical reasons for opposing hard technology, such as nuclear power, and supporting soft technology, such as rooftop solar. The positions taken by various advocates are imbedded in an historical and philosophical perspective. Suggestions are brought forth for improving the level of the debate over the various issues discussed.

  4. Function Blocking Antibodies to Neuropilin-1 Generated from a Designed Human Synthetic Antibody Phage Library

    Microsoft Academic Search

    Wei-Ching Liang; Mark S. Dennis; Scott Stawicki; Yvan Chanthery; Qi Pan; Yongmei Chen; Charles Eigenbrot; JianPing Yin; Alexander W. Koch; Xiumin Wu; Napoleone Ferrara; Anil Bagri; Marc Tessier-Lavigne; Ryan J. Watts; Yan Wu

    2007-01-01

    Non-immune (naïve) antibody phage libraries have become an important source of human antibodies. The synthetic phage antibody library described here utilizes a single human framework with a template containing human consensus complementarity-determining regions (CDRs). Diversity of the libraries was introduced at select CDR positions using tailored degenerate and trinucleotide codons that mimic natural human antibodies. Neuropilin-1 (NRP1), a cell-surface receptor

  5. Serum Antibodies Positivity to 12 Helicobacter pylori Virulence Antigens in Patients with Benign or Malignant Gastroduodenal Diseases – Cross-sectional Study

    PubMed Central

    Filipec Kanižaj, Tajana; Kati?i?, Miroslava; Prese?ki, Vladimir; Gašparov, Slavko; Coli? Cvrlje, Vesna; Kolari?, Branko; Mrzljak, Anna

    2009-01-01

    Aim To investigate the association of gastric histological and endoscopic findings in patients with Helicobacter pylori (H. pylori), according to presence of seropositivity to 12 bacterial virulence antigens. Methods This is a cross-sectional single-center study of 360 consecutive outpatients referred in the period of one year to upper gastrointestinal endoscopy because of dyspeptic complaints. Patients sera were tested by Western blot method to determine the presence of serum antibodies to bacterial virulence antigens – p120 (CagA – cytotoxin-associated antigen), p95 (VacA – vacuolating cytotoxin), p67 (FSH – flagellar sheath protein), p66 (UreB – urease enzyme heavy subunit), p57 (HSP homologue – heath shock protein homologue), p54 (flagellin), p33, p30 (OMP – outer membrane protein), p29 (UreA – urease enzyme light subunit), p26, p19, and p17. Upper gastrointestinal endoscopy was performed, endoscopic diagnosis recorded, and 4 mucosal biopsy samples were obtained and assessed according to Updated Sydney protocol. Results The sera of 207 patients were analyzed. Thirty patients had gastric adenocarcinoma, 126 peptic ulcers, and 51 normal finding. p120 (CagA) seropositivity was significantly more often present in patients with higher activity grade in the antrum (P?=?0.025), p30 in patients with greater inflammation in the antrum (P?=?0.025) and the corpus (P?=?0.010), p33 in patients with greater inflammation in the corpus (P?=?0.050), and p19 (OMP) in patients with lower intestinal metaplasia grades in the corpus (P?=?0.025). Seroreactivity to all other bacterial proteins showed no association with the histological status of the stomach mucosa. Except for the seropositivity to protein p95 (VacA), which was more often present in patients with duodenal ulcer (P?=?0.006), there was no difference in seroreactivity to other bacterial proteins and upper gastrointestinal endoscopic findings. Conclusions p120 (CagA), p33, p 30 (OMP), and p19 (OMP) seropositivity was more often present in patients with higher grades of the histological parameters of gastritis and seropositivity to protein p95 (VacA) with endoscopic presence of duodenal ulcer. Histological parameters of gastritis are more associated with bacterial virulence than endoscopic findings. PMID:19399945

  6. Association of Valine and Leucine at HLA–DRB1 Position 11 With Radiographic Progression in Rheumatoid Arthritis, Independent of the Shared Epitope Alleles but Not Independent of Anti–Citrullinated Protein Antibodies

    PubMed Central

    van Steenbergen, H. W.; Raychaudhuri, S.; Rodríguez-Rodríguez, L.; Rantapää-Dahlqvist, S.; Berglin, E.; Toes, R. E. M.; Huizinga, T. W. J.; Fernández-Gutiérrez, B.; Gregersen, P. K.; van der Helm-van Mil, A. H. M.

    2015-01-01

    Objective For decades it has been known that the HLA–DRB1 shared epitope (SE) alleles are associated with an increased risk of development and progression of rheumatoid arthritis (RA). Recently, the following variations in the peptide-binding grooves of HLA molecules that predispose to RA development have been identified: Val and Leu at HLA–DRB1 position 11, Asp at HLA–B position 9, and Phe at HLA–DPB1 position 9. This study was undertaken to investigate whether these variants are also associated with radiographic progression in RA, independent of SE and anti–citrullinated protein antibody (ACPA) status. Methods A total of 4,911 radiograph sets from 1,878 RA patients included in the Leiden Early Arthritis Clinic (The Netherlands), Umeå (Sweden), Hospital Clinico San Carlos–Rheumatoid Arthritis (Spain), and National Data Bank for Rheumatic Diseases (US) cohorts were studied. HLA was imputed using single-nucleotide polymorphism data from an Immunochip, and the amino acids listed above were tested in relation to radiographic progression per cohort using an additive model. Results from the 4 cohorts were combined in inverse-variance weighted meta-analyses using a fixed-effects model. Analyses were conditioned on SE and ACPA status. Results Val and Leu at HLA–DRB1 position 11 were associated with more radiographic progression (meta-analysis P = 5.11 × 10?7); this effect was independent of SE status (meta-analysis P = 0.022) but not independent of ACPA status. Phe at HLA–DPB1 position 9 was associated with more severe radiographic progression (meta-analysis P = 0.024), though not independent of SE status. Asp at HLA–B position 9 was not associated with radiographic progression. Conclusion Val and Leu at HLA–DRB1 position 11 conferred a risk of a higher rate of radiographic progression independent of SE status but not independent of ACPA status. These findings support the relevance of these amino acids at position 11. PMID:25580908

  7. Antibody production

    Microsoft Academic Search

    John R. Birch; Andrew J. Racher

    2006-01-01

    The clinical and commercial success of monoclonal antibodies has led to the need for very large-scale production in mammalian cell culture. This has resulted in rapid expansion of global manufacturing capacity [1], an increase in size of reactors (up to 20,000 L) and a greatly increased effort to improve process efficiency with concomitant manufacturing cost reduction. This has been particularly successful

  8. Antibody to dihydropyridine calcium entry blockers

    SciTech Connect

    Thayer, S.; Minaskanian, G.; Fairhurst, A.

    1986-03-05

    Antibodies that recognize dihydropyridine calcium entry blockers were elicited from rabbits. A sensitive and specific radioimmunoassay for dihydropyridines was developed and its specificity compared to the DHP binding site in skeletal muscle membranes. The antibody bound (/sup 3/H)nitrendipine with a higher affinity (K/sub D/ = 0.155 nM) than did the DHP receptor of skeletal muscle (K/sub D/ = 1-3 nM). However, in contrast to the DHP receptor, the antibody recognized only those DHP drugs with meta-nitrophenyl substituents at the 4-position on the DHP ring, and thus reflected the meta position of the nitro group on the DHP hapten used as an antigen. Both the antibody and the receptor exhibited stereospecificity, with each site recognizing the (+) isomer of nicardipine as the more potent. This antibody should prove useful in the studies of some potentially irreversible DHP molecules and for use in the production of anti-idotype antibodies.

  9. A monoclonal antibody that recognizes a phosphorylated epitope stains lampbrush chromosome loops and small granules in the amphibian germinal vesicle

    PubMed Central

    1990-01-01

    An mAb library was produced against proteins from the germinal vesicle (GV) of the frog Xenopus laevis; mAb 104 was selected from this library on the basis of its immunofluorescent staining of lampbrush chromosome loops. Chromosomes from several species of frogs and salamanders stained equally well. The antibody also stained the surface of numerous small granules in the GV nucleoplasm. The interior of the same granules was stained by antibodies against small nuclear ribonucleoproteins (snRNPs). mAb 104 also stained somatic nuclei from many vertebrate and invertebrate species, usually in a finely punctate pattern similar to that described for anti-snRNP and other antinuclear antibodies. The staining of somatic nuclei was much stronger during the mitotic stages than during interphase. Immunoblot analysis showed that mAb 104 recognizes a phosphorylated epitope. PMID:1703534

  10. Antibody Engineering and Therapeutics

    PubMed Central

    Almagro, Juan Carlos; Gilliland, Gary L; Breden, Felix; Scott, Jamie K; Sok, Devin; Pauthner, Matthias; Reichert, Janice M; Helguera, Gustavo; Andrabi, Raiees; Mabry, Robert; Bléry, Mathieu; Voss, James E; Laurén, Juha; Abuqayyas, Lubna; Barghorn, Stefan; Ben-Jacob, Eshel; Crowe, James E; Huston, James S; Johnston, Stephen Albert; Krauland, Eric; Lund-Johansen, Fridtjof; Marasco, Wayne A; Parren, Paul WHI; Xu, Kai Y

    2014-01-01

    The 24th Antibody Engineering & Therapeutics meeting brought together a broad range of participants who were updated on the latest advances in antibody research and development. Organized by IBC Life Sciences, the gathering is the annual meeting of The Antibody Society, which serves as the scientific sponsor. Preconference workshops on 3D modeling and delineation of clonal lineages were featured, and the conference included sessions on a wide variety of topics relevant to researchers, including systems biology; antibody deep sequencing and repertoires; the effects of antibody gene variation and usage on antibody response; directed evolution; knowledge-based design; antibodies in a complex environment; polyreactive antibodies and polyspecificity; the interface between antibody therapy and cellular immunity in cancer; antibodies in cardiometabolic medicine; antibody pharmacokinetics, distribution and off-target toxicity; optimizing antibody formats for immunotherapy; polyclonals, oligoclonals and bispecifics; antibody discovery platforms; and antibody-drug conjugates. PMID:24589717

  11. 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, ?2-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 IgG anti-CMV were associated with production of lupus-related autoantibodies to RNA or DNA–protein complex (P = 0.0077). Conclusions Our findings suggest a potential role of CMV in regulation of autoantibodies to snRNPs and may provide a unique insight to understand the pathogenesis. PMID:19232124

  12. Antibody production.

    PubMed

    Birch, John R; Racher, Andrew J

    2006-08-01

    The clinical and commercial success of monoclonal antibodies has led to the need for very large-scale production in mammalian cell culture. This has resulted in rapid expansion of global manufacturing capacity [1], an increase in size of reactors (up to 20,000 L) and a greatly increased effort to improve process efficiency with concomitant manufacturing cost reduction. This has been particularly successful in the upstream part of the process where productivity of cell cultures has improved 100 fold in the last 15 years. This success has resulted from improvements in expression technology and from process optimisation, especially the development of fed-batch cultures. In addition to improving process/cost efficiencies, a second key area has been reducing the time taken to develop processes and produce the first material required for clinical testing and proof-of-principle. Cell line creation is often the slowest step in this stage of process development. This article will review the technologies currently used to make monoclonal antibodies with particular emphasis on mammalian cell culture. Likely future trends are also discussed. PMID:16822577

  13. Overcoming a "probable" diagnosis in antimitochondrial antibody negative primary biliary cirrhosis: study of 100 sera and review of the literature.

    PubMed

    Bizzaro, Nicola; Covini, Giovanni; Rosina, Floriano; Muratori, Paolo; Tonutti, Elio; Villalta, Danilo; Pesente, Fiorenza; Alessio, Maria Grazia; Tampoia, Marilina; Antico, Antonio; Platzgummer, Stefan; Porcelli, Brunetta; Terzuoli, Lucia; Liguori, Marco; Bassetti, Danila; Brusca, Ignazio; Almasio, Piero L; Tarantino, Giuseppe; Bonaguri, Chiara; Agostinis, Paolo; Bredi, Elena; Tozzoli, Renato; Invernizzi, Pietro; Selmi, Carlo

    2012-06-01

    Serum anti-mitochondrial antibodies (AMA) are the serological hallmark of primary biliary cirrhosis (PBC), yet up to 15% of PBC sera are AMA negative at routine indirect immunofluorescence (IIF) while being referred to as "probable" cases. The diagnostic role of PBC-specific antinuclear antibodies (ANA) remains to be determined. We will report herein data on the accuracy of new laboratory tools for AMA and PBC-specific ANA in a large series of PBC sera that were AMA-negative at IIF. We will also provide a discussion of the history and current status of AMA detection methods. We included IIF AMA-negative PBC sera (n=100) and sera from patients with other chronic liver diseases (n=104) that had been independently tested for IIF AMA and ANA; sera were blindly tested with an ELISA PBC screening test including two ANA (gp210, sp100) and a triple (pMIT3) AMA recombinant antigens. Among IIF AMA-negative sera, 43/100 (43%) manifested reactivity using the PBC screening test. The same test was positive for 6/104 (5.8%) control sera. IIF AMA-negative/PBC screen-positive sera reacted against pMIT3 (11/43), gp210 (8/43), Sp100 (17/43), both pMIT3 and gp210 (1/43), or both pMIT3 and Sp100 (6/43). Concordance rates between the ANA pattern on HEp-2 cells and specific Sp100 and gp210 ELISA results in AMA-negative subjects were 92% for nuclear dots and Sp100 and 99% for nuclear rim and gp210. Our data confirm the hypothesis that a substantial part of IIF AMA-negative (formerly coined "probable") PBC cases manifest disease-specific autoantibodies when tested using newly available tools and thus overcome the previously suggested diagnostic classification. As suggested by the recent literature, we are convinced that the proportion of AMA-negative PBC cases will be significantly minimized by the use of new laboratory methods and recombinant antigens. PMID:21188646

  14. Aged venous thrombi: radioimmunoimaging with fibrin-specific monoclonal antibody

    SciTech Connect

    Rosebrough, S.F.; Grossman, Z.D.; McAfee, J.G.; Kudryk, B.J.; Subramanian, G.; Ritter-Hrncirik, C.A.; Witanowski, L.S.; Tillapaugh-Fay, G.; Urrutia, E.

    1987-02-01

    Radioimmunoimaging of fresh canine venous thrombi with a murine monoclonal antibody specific for human and dog fibrin has been reported. Successful imaging of canine deep venous thrombi 1, 3, and 5 days old at the time of antibody injection is reported. Images were positive in all dogs, and the uptake of fibrin-specific antibody was equivalent to that of fresh thrombi.

  15. Technetium-99m-labeled monoclonal antibodies for immunoscintigraphy. Simplified preparation and evaluation.

    PubMed

    Thakur, M L; DeFulvio, J D

    1991-03-21

    Ascorbic acid incubated with monoclonal antibodies (22 degrees C, 60 min, pH 6.5) at a molar ratio of 3500:1, reduced 2.7 +/- 0.2% of the available disulfides to sulfhydryl groups that strongly bind 99mTc, and provided greater than 95% labeling efficiency for several IgM, IgG and F(ab')2 antibodies. The colloid formation was consistently less than 3% and the stability of the tracer when challenged with DTPA and cysteine was excellent. The immunospecificity of labeled antibodies as determined by immobilized specific antigen assay was 84 +/- 1% for IgM and 82.6 +/- 1.1% for IgG antibodies. For in vivo evaluation in mice bearing experimental abscesses and tumors, corresponding 125I-labeled antibodies served as controls. The liver uptake was similar (P = 0.76 and P = 0.12) for 99mTc or 125I labeled antinuclear antibody TNT-1 in mice bearing abscesses as well as for 99mTc-TNT-1-F(ab')2 and 125I-TNT-1-F(ab')2 in mice bearing tumors. Higher but statistically insignificant (P = 0.08, 0.18, and 0.73) urinary excretion was noted for 99mTc-antibodies. For corresponding 99mTc- and 125I-labeled antibodies, the abscess to muscle ratios (3.3 +/- 0.5 vs. 3.4 +/- 0.8) and tumor to muscle ratios (10.04 +/- 4.4 vs. 10.54 +/- 3.0) were similar. The high 99mTc-TNT-1-F(ab')2 uptake permitted excellent scintigraphic visualization of tumors whereas the nonspecific 99mTc-HSA did not (tumor/muscle ratio: 2.4 +/- 0.3). This method is simple, reliable, and adaptable to an instant labeling technique. PMID:2013698

  16. Cross-reactive and pre-existing antibodies to therapeutic antibodies-Effects on treatment and immunogenicity.

    PubMed

    van Schie, Karin A; Wolbink, Gerrit-Jan; Rispens, Theo

    2015-07-01

    The potential for immunogenicity is an ever-present concern during the development of biopharmaceuticals. Therapeutic antibodies occasionally elicit an antibody response in patients, which can result in loss of response or adverse effects. However, antibodies that bind a drug are sometimes found in pre-treatment serum samples, with the amount depending on drug, assay, and patient population. This review summarizes published data on pre-existing antibodies to therapeutic antibodies, including rheumatoid factors, anti-allotype antibodies, anti-hinge antibodies, and anti-glycan antibodies. Unlike anti-idiotype antibodies elicited by the drug, pre-formed antibodies in general appear to have little consequences during treatment. In the few cases where (potential) clinical consequences were encountered, antibodies were characterized and found to bind a distinct, unusual epitope of the therapeutic. Immunogenicity testing strategies should therefore always include a proper level of antibody characterization, especially when pre-formed antibodies are present. This minimizes false-positives, particularly due to rheumatoid factors, and helps to judge the potential threat in case a genuine pre-dose antibody reactivity is identified. PMID:25962087

  17. Antibody profiling sensitivity through increased reporter antibody layering

    DOEpatents

    Apel, William A.; Thompson, Vicki S.

    2013-02-26

    A method for analyzing a biological sample by antibody profiling for identifying forensic samples or for detecting the presence of an analyte. In an embodiment of the invention, the analyte is a drug, such as marijuana, Cocaine (crystalline tropane alkaloid), methamphetamine, methyltestosterone, or mesterolone. The method comprises attaching antigens to a surface of a solid support in a preselected pattern to form an array wherein locations of the antigens are known; contacting the array with the biological sample such that a portion of antibodies in the sample reacts with and binds to the antigens in the array to form immune complexes; washing away antibodies that do form immune complexes; and detecting the immune complexes, to form an antibody profile. Forensic samples are identified by comparing a sample from an unknown source with a sample from a known source. Further, an assay, such as a test for illegal drug use, can be coupled to a test for identity such that the results of the assay can be positively correlated to the subject's identity.

  18. Antibody profiling sensitivity through increased reporter antibody layering

    DOEpatents

    Apel, William A.; Thompson, Vicki S

    2010-04-13

    A method for analyzing a biological sample by antibody profiling for identifying forensic samples or for detecting the presence of an analyte. In an embodiment of the invention, the analyte is a drug, such as marijuana, Cocaine (crystalline tropane alkaloid), methamphetamine, methyltestosterone, or mesterolone. The method comprises attaching antigens to a surface of a solid support in a preselected pattern to form an array wherein locations of the antigens are known; contacting the array with the biological sample such that a portion of antibodies in the sample reacts with and binds to the antigens in the array to form immune complexes; washing away antibodies that do form immune complexes; and detecting the immune complexes, to form an antibody profile. Forensic samples are identified by comparing a sample from an unknown source with a sample from a known source. Further, an assay, such as a test for illegal drug use, can be coupled to a test for identity such that the results of the assay can be positively correlated to the subject's identity.

  19. Peptide antibodies and their use in detecting oncogene products

    SciTech Connect

    Wong, G.L.; Arnheim, N.; McCormick, F.P.; Wong, G.L.; Clark, R.; Arnheim, N.; Nitecki, D.E.

    1989-01-17

    A polyclonal antibody preparation is described. It binds selectively to a characteristic marker epitope encompassing amino acid position 12 of an activated form of p21 protein, wherein the polyclonal antibody preparation is specific for a particular polyclonal antibody amino acid at position 12 and does not bind the p21 protein encoded by the corresponding proto-oncogene. A method for detecting an activated form of p21 protein is also described. It is encoded by an oncogene in a cellular sample of a patient which protein has a characteristic marker epitope encompassing amino acid position 12 which is not present in the p21 protein encoded by the corresponding proto-oncogene and which is not exposed in the undenatured protein. The method comprises: (a) treating the sample with a protein denaturing agent that causes the epitope to be exposed and does not substantially inhibit binding of an antibody to the epitope of the protein, (b) incubating the sample with the antibody under conditions that permit the binding of the antibody preparation to the epitope, (c) incubating the sample with a labeled antibody which binds specifically to the antibody employed in step (b), (d) washing the incubated sample to remove unbound labeled antibody, and (e) detecting the presence of labelled immune complexes of the epitope with the antibodies employed in steps (b) and (c).

  20. Passive West Nile virus antibody transfer from maternal Eastern screech-owls (Megascops asio) to progeny.

    PubMed

    Hahn, D C; Nemeth, Nicole M; Edwards, Eric; Bright, Patricia R; Komar, Nicholas

    2006-09-01

    Transovarial antibody transfer in owls has not been demonstrated for West Nile virus (WNV). We sampled chicks from captive adult WNV-antibody-positive Eastern Screech-Owls (Megascops asio) to evaluate the prevalence of transovarial maternal antibody transfer, as well as titers and duration of maternal antibodies. Twenty-four owlets aged 1 to 27 days old circulated detectable antibodies with neutralizing antibody titers ranging from 20 to 1600 (median 1:40). Demonstrating that WNV antibodies are passively transferred transovarially is important for accurate interpretation of serologic data from young birds. PMID:17039850

  1. Passive West Nile virus antibody transfer from maternal Eastern Screech-Owls (Megascops asio) to progeny

    USGS Publications Warehouse

    Hahn, D.C.; Nemeth, N.M.; Edwards, E.; Bright, P.R.; Komar, N.

    2006-01-01

    Transovarial antibody transfer in owls has not been demonstrated for West Nile virus (WNV). We sampled chicks from captive adult WNV-antibody-positive Eastern Screech-Owls (Megascops asio) to evaluate the prevalence of transovarial maternal antibody transfer, as well as titers and duration of maternal antibodies. Twenty-four owlets aged 1 to 27 days old circulated detectable antibodies with neutralizing antibody titers ranging from 20 to 1600 (median 1:40). Demonstrating that WNV antibodies are passively transferred transovarially is important for accurate interpretation of serologic data from young birds.

  2. [Characterisation of a monoclonal antibody against Trypanosoma evansi and its application for detecting circulating antibodies].

    PubMed

    Monzón, C M

    2006-12-01

    Monoclonal antibodies were obtained against Trypanosoma evansi. The 2-4F6 IgM monoclonal antibody (Mab) was chosen for the study because of its ability to detect antigens and its specificity (as it did not recognise T. cruzi, T. equiperdum, Babesia equi or B. caballi). The immunoblot test revealed that the 2-4F6 IgM Mab recognises epitopes in two antigenic bands, one measuring 85 kDa and the other 122 kDa. An immunoassay for antigen detection in serum using polyclonal antibodies for capture, the Mab 2-4F6 as primary antibody and an antimouse IgM as secondary antibody gave positive results in 10 of the 11 equidae infected with T. evansi, whereas 20 controls gave negative results. These research results show that the Mab 2-4F6 and the antigen it recognises are useful in identifying equidae infected with T. evansi. PMID:17361770

  3. Counterimmunoelectrophoresis, ELISA and immunoblotting detection of anti-Ro/SSA antibodies in subacute cutaneous lupus erythematosus. A comparative study.

    PubMed

    Parodi, A; Drosera, M; Barbieri, L; Rebora, A

    1998-01-01

    Patients with subacute cutaneous lupus erythematosus (SCLE) have circulating antibodies to Ro/SSA directed to two antigenically distinct ribonucleoproteins of 60 kDa and 52 kDa. Three laboratory tests may be used to detect anti-Ro/SSA antibodies: counterimmunoelectrophoresis (CIE), enzyme-linked immunosorbent assay (ELISA) and immunoblotting (IB). Their relative efficacy and clinical correlations were ascertained. We determined anti-Ro/SSA antibodies with CIE, with two different ELISA methods (ELISA 1 and 2) and with IB in 29 SCLE patients. Anti-52 kDa and -60 kDa Ro/SSA antibodies were also assayed with IB. In addition, we determined antinuclear antibodies with indirect immunofluorescence, anti-Sm, anti-RNP, anti-La/SSB and anti-Ro/SSA antibodies with CIE and ELISA, and anti-nDNA and cardiolipin antibodies using an ELISA method. CIE detected anti-Ro/SSA antibodies in 22 patients while ELISA 1 and 2 did so in 17 and 18 patients, respectively. In five patients, IB revealed a reactivity to 60 kDa polypeptides and in two, a reactivity to 52 kDa polypeptides. Of these seven patients, four had a myocardial infarction. Of these, two reacted to the 52 kDa antigen and two to the 60 kDa antigen. A combination of techniques was often needed to detect all specificities. ELISA proved to be very specific and sensitive. The IB technique detected a group of patients with myocardial infarction. A case-control study is needed to confirm the data of cardiac involvement. PMID:9536232

  4. Expression studies of catalytic antibodies.

    PubMed Central

    Ulrich, H D; Patten, P A; Yang, P L; Romesberg, F E; Schultz, P G

    1995-01-01

    We have examined the positive influence of human constant regions on the folding and bacterial expression of active soluble mouse immunoglobulin variable domains derived from a number of catalytic antibodies. Expression yields of eight hybridoma- and myeloma-derived chimeric Fab fragments are compared in both shake flasks and high density fermentations. In addition the usefulness of this system for the generation of in vivo expression libraries is examined by constructing and expressing combinations of heavy and light chain variable regions that were not selected as a pair during an immune response. A mutagenesis study of one of the recombinant catalytic Fab fragments reveals that single amino acid substitutions can have dramatic effects on the expression yield. This system should be generally applicable to the production of Fab fragments of catalytic and other hybridoma-derived antibodies for crystallographic and structure-function studies. Images Fig. 3 Fig. 4 PMID:8524873

  5. Antibody response to Epstein-Barr virus in infectious mononucleosis.

    PubMed Central

    Nikoskelainen, J; Hänninen, P

    1975-01-01

    Altogether 171 serum specimens from 58 patients with heterophil antibody-positive infectious monomucleosis were studied for antibody response to Epstein-Barr virus (EBV). The sera were tested for fluorescent immunoglobulin G (IgG) and IgM gel-precipitating (GP) and complement-fixing (CF) antibodies to EBV. All 58 patients had IgG and IgM antibodies to EBV. Both IgG and IgM antibodies developed rapidly; the IgM antibodies disappeared within 8 to 10 weeks, whereas the IgG antibodies remained at an almost constant level. The development of IgG antibodies was so rapid that a fourfold or greater rise in titers was noted only in 22% of the patients. Both GP and CF antibodies to EBV (crude P3HR-1 Burkitt cell antigen) developed slowly; the mean titers kept rising for more than 12 weeks. The micro GP technique seemed to be more sensitive than the CF method, because 86% of the patients with infectious mononucleosis had GP antibodies compared with 72% having CF antibodies. In patients with infectious mononucleosis, a seroconversion or significant rise in GP antibodies was noted in 57%, whereas only 19% had a similar change in CF antibodies. The most promising of these antibody assays in the diagnosis of recent infections was the EBV-specific IgM antibody technique, which enables one to make the diagnosis on the basis of only one serum specimen. In cases where the acute-phase serum specimen is missing, the diagnosis can be made later by using the GP and CF techniques. Images PMID:163790

  6. Antibodies to citrullinated peptides in tuberculosis.

    PubMed

    Lima, I; Oliveira, R C; Atta, A; Marchi, S; Barbosa, L; Reis, E; Reis, M G; Santiago, M B

    2013-05-01

    Rheumatoid arthritis (RA) is an autoimmune disease characterized by symmetric polyarthritis, rheumatoid factor (RF) positivity, and bone erosions. Recently, research has been conducted on anti-citrullinated peptide antibodies (ACPAs) to which there are greater sensitivity and specificity than RF. However, these antibodies have also been described in infectious diseases, particularly tuberculosis (TB), placing the high specificity of the test in doubt. The aim of this research was to study the prevalence of ACPAs in TB, RA, and healthy controls. Patients with bacteriologically confirmed pulmonary tuberculosis, RA (ACR criteria), in addition to healthy controls were included. ACPAs were researched by: anti-cyclic citrullinated peptide (CCP), anti-modified citrullinated vimentin (MCV), and RF by ELISA. The study was conducted in 50 TB patients, 50 with RA, and 20 controls. Anti-CCP antibodies were found in 39 (78 %) of the RA patients (median titer, 128 U), whereas anti-MCV antibodies were found in 25 (50 %). Of the patients with TB, two (4 %) had positivity for anti-CCP and anti-MCV and no patient in the control group tested positive for these antibodies. Sensitivity of anti-CCP for RA was 78 % (confidence interval (CI), 63 to 88 %) and specificity was 97 % (CI, 89 to 99 %) while the sensitivity of anti-MCV was 50 % (CI, 35-64 %) and specificity was 97 % (CI, 89 to 99 %). RF was positive in 40 samples (80 %) of RA, in 30 (60 %) of TB, and in 1 (5 %) of the controls. Our findings showed high sensitivity of anti-CCP and high specificity of both anti-CCP and anti-MCV antibodies for RA, even in a population with high incidence of tuberculosis. The higher frequency of positivity of ACPA in TB observed in previous studies may be attributed to methodological factors. PMID:23344687

  7. Therapeutic antibody expression technology.

    PubMed

    Chadd, H E; Chamow, S M

    2001-04-01

    With the technological advances made during the past decade, antibodies now represent an important and growing class of biotherapeutics. With the potential new targets resulting from genomics and with methods now in place to make fully human antibodies, the potential of antibodies as valuable therapeutics in oncology, inflammation and cardiovascular disease can be fully realised. Systems to produce these antibodies as full-length molecules and as fragments include expression in both mammalian and bacterial cells grown in bioreactors and in transgenic organisms. Factors including molecular fidelity and the cost of goods are critical in evaluating expression systems. Mammalian cell culture and transgenic organisms show the greatest promise for the expression of full-length, recombinant human antibodies, and bacterial fermentation seems most favorable for the expression of antibody fragments. PMID:11287236

  8. Biobarcodes: Antibodies and Nanosensors

    NSDL National Science Digital Library

    Nanoscale Informal Science Education Network

    2014-06-04

    In this activity/demo, learners investigate biobarcodes, a nanomedical technology that allows for massively parallel testing that can assist with disease diagnosis. Learners define antibodies and learn how each antibody binds to a unique protein. Learners also discover how biobarcoding uses nanoparticles, antibodies, DNA and magnetism to detect diseases earlier than we could detect before. Learners assemble a jigsaw puzzle that models how biobarcodes work.

  9. Red Blood Cell Antibody Identification

    MedlinePLUS

    ... Antibody ID, RBC; RBC Ab ID Formal name: Red Blood Cell Antibody Identification Related tests: Direct Antiglobulin ... else I should know? How is it used? Red blood cell antibody identification is used as a ...

  10. Circulating Antibodies in Dermatophytosis

    Microsoft Academic Search

    Sarah F. Grappel; F. Blank; C. T. Bishop

    1972-01-01

    Antibodies to dermatophytes (M. audouinii, T. mentagrophytes var. granulosum and var. interdigitale, T. rubrum, and T. tonsurans) were detected in sera of patients with tinea capitis and tinea corporis by charcoal agglutination tests, immunodiffusion analysis, and complement fixation tests. In tinea capitis, these antibodies were not confined to patients with deep-seated, inflammatory infections. The zoophilic variety of T. mentagrophytes induced

  11. Antibodies in Plants

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The expression of antibodies in plants has several promising applications that are currently being developed. Plants are being considered for the large scale production of antibodies needed for medical purposes. The benefit of using plants is that they are able to perform post-translational modifi...

  12. Association of anti-phospholipid antibodies with connective tissue diseases

    PubMed Central

    Rai, Reena; Swetha, T.

    2015-01-01

    Background: The antiphospholipid antibodies (APLA) are directed against phospholipids and their binding proteins and are frequently found in association with connective tissue disorders. Systemic lupus erythematoses (SLE) with APLA may cause a diagnostic dilemma as there are several manifestations like haemolytic anemia, thrombocytopenia, neurologic manifestations, leg ulcerations, serositis proteinuria which overlap in both these conditions. We conducted a study to find out the association of antiphospholipid antibodies with connective tissue diseases and compared the clinical and laboratory parameters between antiphoshpolipid antibody positive and antiphoshpolipid antibody negative group. Materials and Methods: This study was carried out in 102 patients diagnosed with connective tissue diseases. APLA testing was done at baseline and for those positive, the test was repeated after 12 weeks. Results: 14.7 % of patients with connective tissue diseases tissue had positive antiphoshpolipid antibodies. Positive antiphoshpolipid antibody was detected in 73.3% of patients with SLE group, 13.3% of patients with mixed connective tissue disease (MCTD) and 13.3% of patients with systemic sclerosis. APLA positivity was seen in SLE patients with leg ulcers (87.2%), neurologic manifestation (72.7%), hemolytic anemia (62.3%), thrombocytopenia (72.7%), serositis (27.8%) and proteinuria(19.6%). Conclusions: Antiphoshpolipid antibodies should be tested in all patients with connective tissue disease. PMID:25821728

  13. A dichromatic latex agglutination test for semiquantitative detection of antibodies to Toxoplasma gondii

    Microsoft Academic Search

    E. H Frost

    1995-01-01

    A dichromatic latex agglutination test was employed to measure antibodies to Toxoplasma gondii in 160 sera. Reactions were deemed negative, weak positive, positive or strong positive on the basis of rapidity of agglutination and\\/or colour change. This semiquantitative determination agreed well with antibody titres measured by immunofluorescence.

  14. Antibodies for biodefense

    PubMed Central

    Froude, Jeffrey W; Stiles, Bradley; Pelat, Thibaut

    2011-01-01

    Potential bioweapons are biological agents (bacteria, viruses and toxins) at risk of intentional dissemination. Biodefense, defined as development of therapeutics and vaccines against these agents, has seen an increase, particularly in the US, following the 2001 anthrax attack. This review focuses on recombinant antibodies and polyclonal antibodies for biodefense that have been accepted for clinical use. These antibodies aim to protect against primary potential bioweapons or category A agents as defined by the Centers for Disease Control and Prevention (Bacillus anthracis, Yersinia pestis, Francisella tularensis, botulinum neurotoxins, smallpox virus and certain others causing viral hemorrhagic fevers) and certain category B agents. Potential for prophylactic use is presented, as well as frequent use of oligoclonal antibodies or synergistic effect with other molecules. Capacities and limitations of antibodies for use in biodefense are discussed, and are generally applicable to the field of infectious diseases. PMID:22123065

  15. Generation of neutralising antibodies against porcine endogenous retroviruses (PERVs)

    SciTech Connect

    Kaulitz, Danny; Fiebig, Uwe; Eschricht, Magdalena; Wurzbacher, Christian; Kurth, Reinhard; Denner, Joachim, E-mail: DennerJ@rki.d

    2011-03-01

    Antibodies neutralising porcine endogenous retroviruses (PERVs) were induced in different animal species by immunisation with the transmembrane envelope protein p15E. These antibodies recognised epitopes, designated E1, in the fusion peptide proximal region (FPPR) of p15E, and E2 in the membrane proximal external region (MPER). E2 is localised in a position similar to that of an epitope in the transmembrane envelope protein gp41 of the human immunodeficiency virus-1 (HIV-1), recognised by the monoclonal antibody 4E10 that is broadly neutralising. To detect neutralising antibodies specific for PERV, a novel assay was developed, which is based on quantification of provirus integration by real-time PCR. In addition, for the first time, highly effective neutralising antibodies were obtained by immunisation with the surface envelope protein of PERV. These data indicate that neutralising antibodies can be induced by immunisation with both envelope proteins.

  16. Sensitivity of HIV antibody detection in saliva.

    PubMed

    Stark, K; Warnecke, C; Brinkmann, V; Gelderblom, H R; Bienzle, U; Pauli, G

    1993-07-01

    To assess the sensitivity and specificity of HIV antibody detection in saliva we tested matched serum and saliva samples from HIV-infected and uninfected individuals. Saliva specimens were collected by two different devices of the Salivette system and stored at different temperatures. Samples were tested for HIV antibodies by two commercially available enzyme-linked immunosorbent assays (ELISAs; Wellcome, Biotest). HIV antibodies were detected in 98.5% (Wellcome) and 97.8% (Biotest) of the saliva samples (standard Salivettes) from 135 seropositive individuals. Using the Salivettes flavoured with citric acid the sensitivity was only 22.9%. No reactions in ELISA were found in saliva from HIV-seronegative individuals. Salivary HIV-specific IgA was detected in 90% of seropositive individuals. All positive saliva samples stored at room temperature were still reactive after 20 days; of those stored at 37 degrees C, 23 out of 24 were positive when retested on day 5. Sensitivity of HIV antibody detection in saliva samples dried onto filter paper was 100% when a minimum of 100 microliters of saliva was used. HIV antibody testing in saliva is an efficient tool for large scale epidemiological studies when standard Salivettes are used for sample collection. Saliva samples can be stored in Salivettes or dried onto filter paper for several days at room temperature and under tropical conditions (37 degrees C). PMID:8232068

  17. Antibodies to Neospora caninum in the blood of European bison ( Bison bonasus bonasus L.) living in Poland

    Microsoft Academic Search

    W. Cabaj; B. Moskwa; K. Pastusiak; J. Gill

    2005-01-01

    The prevalence of antibodies to Neospora caninum was examined in European bison (Bison bonasus bonasus L.) living in free and fenced areas in Poland. Sera of 320 European bison, different ages and sexes, from breeding areas in Poland were tested for N. caninum antibodies using ELISA test. Positive antibody responses were found in 23 bison (prevalence 7.3%). Additionally, all positive

  18. Identification of interferon-beta antibodies in a reference laboratory setting: Findings for 1144 consecutive sera

    Microsoft Academic Search

    Harry E. Prince; Mary Lapé-Nixon; Carol Audette; Kenneth Van Horn

    2007-01-01

    Clinical studies demonstrate differences in interferon-beta (IFN?) antibody detection frequencies among multiple sclerosis patients receiving different IFN? products. We sought to determine if these differences are also found when IFN? antibodies are measured in a reference laboratory, where factors normally controlled in clinical studies are unknown. Serum IFN? binding antibodies (BAbs) were quantitated by ELISA; BAbs-positive samples were then tested

  19. A serological survey of domestic poultry in the united kingdom for antibody to chicken anaemia agent

    Microsoft Academic Search

    M. S. McNulty; T. J. Connor; F. McNeilly; K. S. Kirkpatrick; J. B. McFerran

    1988-01-01

    A serological survey for antibody to chicken anaemia agent (CAA) was carried out by indirect immunofluorescence. Antibody to CAA was widespread in broiler breeders and in parent and commercial layers in the UK. Antibody to CAA was also detected in five of 11 specific pathogen?free (SPF) chicken flocks tested; positive flocks included those being used for vaccine production or as

  20. Adenovirus hexon monoclonal antibody that is group specific and potentially useful as a diagnostic reagent.

    PubMed Central

    Cepko, C L; Whetstone, C A; Sharp, P A

    1983-01-01

    A monoclonal antibody to the adenovirus 2 hexon protein was produced and characterized as a group-specific antibody. Positive reactivity in immunoprecipitation, indirect immunofluorescence, and radioimmunoassays was observed with human, canine, swine, bovine, murine, and simian adenoviruses. This monoclonal antibody should provide a specific and sensitive diagnostic reagent for detection of all mammalian adenoviruses. Images PMID:6339552

  1. Role of Maternal Antibody in Natural Infection of Peromyscus maniculatus with Sin Nombre Virus

    Microsoft Academic Search

    MONICA K. BORUCKI; JOHN D. BOONE; JOAN E. ROWE; MARLENE C. BOHLMAN; EDWARD A. KUHN; ROBERT DEBACA; STEPHEN C. S. T. JEOR

    2000-01-01

    Data from naturally infected deer mice (Peromyscus maniculatus) were used to investigate vertical transmis- sion of Sin Nombre virus (SNV) and SNV-specific antibody. The antibody prevalence in juvenile mice (14 g or less) was inversely proportional to the mass of the animal, with juvenile deer mice weighing less than 11 g most likely to be antibody positive (26.9%) and juvenile

  2. Acute antibody-mediated renal allograft rejection associated with HLA-Cw17 antibody

    PubMed Central

    Kuppachi, Sarat

    2012-01-01

    Detection of donor-specific human leukocyte antigen (HLA) antibodies is an important part of diagnosis of antibody-mediated rejection (AMR) in the renal transplant population. Donor-specific antibodies (DSA) against HLA-C, a Class 1 major histocompatibility gene product, are not considered to be of major importance in renal transplant rejection. Typing for HLA-C is not a routine part of pre- and post-transplant evaluation. In roughly 10% of biopsy-proven C4d-positive rejections, DSA are not detected by standard testing protocols. In some of these cases, minor HLA and non-HLA antibodies have been implicated. The role of HLA-C antibodies in this patient group is not clear. We present a patient with acute renal graft dysfunction 21 months post-transplant. The allograft biopsy showed features of AMR with diffuse margination of inflammatory cells and diffuse C4d staining in peritubular capillaries. HLA-Cw17 antibody was detected by single-bead antigen Luminex assay, which was further confirmed by a mock flow crossmatch. This case highlights the importance of checking anti-HLA-Cw antibodies in patients with AMR and no detectable DSA using standard methods.

  3. A novel method of Multiplexed Competitive Antibody Binning for the characterization of monoclonal antibodies.

    PubMed

    Jia, Xiao-Chi; Raya, Robert; Zhang, Li; Foord, Orit; Walker, Wynn L; Gallo, Michael L; Haak-Frendscho, Mary; Green, Larry L; Davis, C Geoffrey

    2004-05-01

    We have developed a novel method of high-throughput Multiplexed Competitive Antibody Binning (MCAB). Using only a small amount of antibody and antigen, this method enables the sorting of a large, complex panel of monoclonal antibodies into different bins based on cross-competition for antigen binding. The MCAB assay builds on Luminex multiplexing bead-based technology to detect antibody competition. Because of its high sensitivity, the MCAB method is immediately applicable after identification of antigen-positive mAbs, providing information useful for advancing mAb candidates into further testing. The MCAB assay also can be used for sorting mAbs into binding groups after screening for functional activity. PMID:15183088

  4. Anti-sulfotyrosine antibodies

    DOEpatents

    Bertozzi, Carolyn R. (Berkeley, CA); Kehoe, John (Saint Davids, PA); Bradbury, Andrew M. (Santa Fe, NM)

    2009-09-15

    The invention provides anti-sulfotyrosine specific antibodies capable of detecting and isolating polypeptides that are tyrosine-sulfated. The sulfotyrosine antibodies and antibody fragments of the invention may be used to discriminate between the non-sulfated and sulfated forms of such proteins, using any number of immunological assays, such ELISAs, immunoblots, Western Blots, immunoprecipitations, and the like. Using a phage-display system, single chain antibodies (scFvs) were generated and screened against tyrosine-sulfated synthetic peptide antigens, resulting in the isolation of scFvs that specifically recognize sulfotyrosine-containing peptides and/or demonstrate sulfotyrosine-specific binding in tyrosine sulfated proteins. The VH and VL genes from one such sulfotyrosine-specific scFv were employed to generate a full length, sulfotyrosine-specific immunoglobulin.

  5. Mining human antibody repertoires

    PubMed Central

    2010-01-01

    Human monoclonal antibodies (mAbs) have become drugs of choice for the management of an increasing number of human diseases. Human antibody repertoires provide a rich source for human mAbs. Here we review the characteristics of natural and non-natural human antibody repertoires and their mining with non-combinatorial and combinatorial strategies. In particular, we discuss the selection of human mAbs from naïve, immune, transgenic and synthetic human antibody repertoires using methods based on hybridoma technology, clonal expansion of peripheral B cells, single-cell PCR, phage display, yeast display and mammalian cell display. Our reliance on different strategies is shifting as we gain experience and refine methods to the efficient generation of human mAbs with superior pharmacokinetic and pharmacodynamic properties. PMID:20505349

  6. Antithyroid antibodies and thyroid function in pediatric patients with celiac disease.

    PubMed

    Kalyoncu, Derya; Urganci, Nafiye

    2015-01-01

    Objective. Aim of the study was to determine the prevalence of autoimmune thyroid disease, persistence of antithyroid antibodies, effect of gluten-free diet, and long-term outcome of thyroid function in pediatric patients with celiac disease (CD). Methods. 67 patients with CD aged from 1 year to 16 years were screened for thyroid antithyroperoxidase, antithyroglobulin and anti-TSH receptor antibodies, serum free triiodothyronine, free thyroxine, and thyroid-stimulating hormone (TSH) at diagnosis and during follow-up. Results. None of the patients had antithyroid antibodies at diagnosis. Antithyroid antibodies became positive in 16.4% of the patients (11/67) 2 to 3 years after the diagnosis of CD. Clinical hypothyroidism was observed only in 3 of 11 CD patients with positive antithyroid antibodies (27.2%). The antithyroid antibodies positive and negative patients did not differ significantly according to compliance to GFD (P > 0.05). A statistically significant difference was observed only in age, in which the patients with positive antithyroid antibodies were younger than the patients with negative antithyroid antibodies (P = 0.004). None of the patients had any change in their thyroid function and antibody profile during their follow-up. Conclusion. Antithyroid antibodies were detected in younger pediatric patients with CD and the prevalence of antithyroid antibodies did not correlate with the duration of gluten intake. PMID:25788942

  7. Structural Comparison of Different Antibodies Interacting with Parvovirus Capsids

    SciTech Connect

    Hafenstein, Susan; Bowman, Valorie D.; Sun, Tao; Nelson, Christian D.S.; Palermo, Laura M.; Chipman, Paul R.; Battisti, Anthony J.; Parrish, Colin R.; Rossmann, Michael G.; Cornell; Purdue

    2009-05-13

    The structures of canine parvovirus (CPV) and feline parvovirus (FPV) complexed with antibody fragments from eight different neutralizing monoclonal antibodies were determined by cryo-electron microscopy (cryoEM) reconstruction to resolutions varying from 8.5 to 18 {angstrom}. The crystal structure of one of the Fab molecules and the sequence of the variable domain for each of the Fab molecules have been determined. The structures of Fab fragments not determined crystallographically were predicted by homology modeling according to the amino acid sequence. Fitting of the Fab and virus structures into the cryoEM densities identified the footprints of each antibody on the viral surface. As anticipated from earlier analyses, the Fab binding sites are directed to two epitopes, A and B. The A site is on an exposed part of the surface near an icosahedral threefold axis, whereas the B site is about equidistant from the surrounding five-, three-, and twofold axes. One antibody directed to the A site binds CPV but not FPV. Two of the antibodies directed to the B site neutralize the virus as Fab fragments. The differences in antibody properties have been linked to the amino acids within the antibody footprints, the position of the binding site relative to the icosahedral symmetry elements, and the orientation of the Fab structure relative to the surface of the virus. Most of the exposed surface area was antigenic, although each of the antibodies had a common area of overlap that coincided with the positions of the previously mapped escape mutations.

  8. An Unusual Mimicker of Systemic Lupus Erythematosus: A Case Report

    PubMed Central

    Aluoch, Aloice O; Farbman, Mathew; Gladue, Heather

    2015-01-01

    We present a case of a 47 year-old African American female with 15 pack-years of tobacco use and heavy alcohol use who presented with arthritis and was found to have a positive antinuclear antibodies (ANA), anti double stranded DNA antibodies (anti-dsDNA), and anti-Sjogren’s syndrome-related antigen A and antigen B (anti-SSA and anti-SSB). She was subsequently found to have a lung adenocarcinoma associated with hypertrophic pulmonary osteoarthropathy (HPO). This demonstrates a case of positive antinuclear antibodies and arthritis in a patient with lung adenocarcinoma, which can be falsely diagnosed as systemic lupus erythematosus.

  9. Antibodies to GABAA receptor ?1 and ?2 subunits

    PubMed Central

    Pettingill, Philippa; Kramer, Holger B.; Coebergh, Jan Adriaan; Pettingill, Rosie; Maxwell, Susan; Nibber, Anjan; Malaspina, Andrea; Jacob, Anu; Irani, Sarosh R.; Buckley, Camilla; Beeson, David; Lang, Bethan; Waters, Patrick

    2015-01-01

    Objective: To search for antibodies against neuronal cell surface proteins. Methods: Using immunoprecipitation from neuronal cultures and tandem mass spectrometry, we identified antibodies against the ?1 subunit of the ?-aminobutyric acid A receptor (GABAAR) in a patient whose immunoglobulin G (IgG) antibodies bound to hippocampal neurons. We searched 2,548 sera for antibodies binding to GABAAR ?, ?, and ? subunits on live HEK293 cells and identified the class, subclass, and GABAAR subunit specificities of the positive samples. Results: GABAAR-Abs were identified in 40 of 2,046 (2%) referred sera previously found negative for neuronal antibodies, in 5/502 (1%) previously positive for other neuronal surface antibodies, but not in 92 healthy individuals. The antibodies in 40% bound to either the ?1 (9/45, 20%) or the ?2 subunits (9/45, 20%) and were of IgG1 (94%) or IgG3 (6%) subclass. The remaining 60% had lower antibody titers (p = 0.0005), which were mainly immunoglobulin M (IgM) (p = 0.0025), and showed no defined subunit specificity. Incubation of primary hippocampal neurons with GABAAR IgG1 sera reduced surface GABAAR membrane expression. The clinical features of 15 patients (GABAAR ?1 n = 6, ?2 n = 5, undefined n = 4) included seizures (47%), memory impairment (47%), hallucinations (33%), or anxiety (20%). Most patients had not been given immunotherapies, but one with new-onset treatment-resistant catatonia made substantial improvement after plasma exchange. Conclusions: The GABAAR ?1 and ?2 are new targets for antibodies in autoimmune neurologic disease. The full spectrum of clinical features, treatment responses, correlation with antibody specificity, and in particular the role of the IgM antibodies will need to be assessed in future studies. PMID:25636713

  10. Salivary IgA antibody to glucosyltransferase in man.

    PubMed

    Smith, D J; Taubman, M A; Ebersole, J L

    1985-08-01

    Parotid salivas of 97 young adults were screened for IgA antibody to glucosyltransferase (GTF) from laboratory strains of Streptococcus mutans (serotypes c and g). Antibody levels to GTF from serotype c positively correlated with levels to serotype g GTF among these salivas. GTF's were prepared from S. mutans obtained from a subset of individuals in this population. All but one saliva showed IgA antibody activity to all of the GTF tested. In addition, the relative magnitude of each subject's antibody level was generally the highest to the GTF from their own S. mutans. Fractions, enriched for IgA by ammonium sulphate precipitation and gel filtration, showed patterns of functional inhibition of GTF activity which were consistent with patterns of IgA antibody activity in ELISA of unfractionated salivas. These data indicate that detectable levels of IgA antibody to S. mutans GTF exist in many young adult salivas, while this IgA antibody activity reacts with GTF from different biotypes, subjects generally show the highest secretory IgA antibody levels to their own GTF, and the relative amount of IgA antibody to GTF and the ability to inhibit GTF activity are roughly correlated. PMID:2931224

  11. Salivary IgA antibody to glucosyltransferase in man.

    PubMed Central

    Smith, D J; Taubman, M A; Ebersole, J L

    1985-01-01

    Parotid salivas of 97 young adults were screened for IgA antibody to glucosyltransferase (GTF) from laboratory strains of Streptococcus mutans (serotypes c and g). Antibody levels to GTF from serotype c positively correlated with levels to serotype g GTF among these salivas. GTF's were prepared from S. mutans obtained from a subset of individuals in this population. All but one saliva showed IgA antibody activity to all of the GTF tested. In addition, the relative magnitude of each subject's antibody level was generally the highest to the GTF from their own S. mutans. Fractions, enriched for IgA by ammonium sulphate precipitation and gel filtration, showed patterns of functional inhibition of GTF activity which were consistent with patterns of IgA antibody activity in ELISA of unfractionated salivas. These data indicate that detectable levels of IgA antibody to S. mutans GTF exist in many young adult salivas, while this IgA antibody activity reacts with GTF from different biotypes, subjects generally show the highest secretory IgA antibody levels to their own GTF, and the relative amount of IgA antibody to GTF and the ability to inhibit GTF activity are roughly correlated. PMID:2931224

  12. Immunotherapy with the trifunctional anti-CD20 x anti-CD3 antibody FBTA05 (Lymphomun) in paediatric high-risk patients with recurrent CD20-positive B cell malignancies.

    PubMed

    Schuster, Friedhelm R; Stanglmaier, Michael; Woessmann, Wilhelm; Winkler, Beate; Siepermann, Meinolf; Meisel, Roland; Schlegel, Paul G; Hess, Jürgen; Lindhofer, Horst; Borkhardt, Arndt; Buhmann, Raymund

    2015-04-01

    Children with B cell malignancies refractory to standard therapy are known to have a poor prognosis and very limited treatment options. Here, we report on the treatment and follow-up of ten patients diagnosed with relapsed or refractory mature B-cell Non Hodgkin Lymphoma (B-NHL), Burkitt leukaemia (B-AL) or pre B-acute lymphoblastic leukaemia (pre B-ALL). All children were treated with FBTA05 (now designated Lymphomun), an anti-CD3 x anti-CD20 trifunctional bispecific antibody (trAb) in compassionate use. Within individual treatment schedules, Lymphomun was applied (a) after allogeneic stem cell transplantation (allo-SCT, n = 6) to induce sustained long-term remission, or (b) stand alone prior to subsequent chemotherapy to eradicate residual disease before allo-SCT (n = 4). Nine of ten children displayed a clinical response: three stable diseases (SD), one partial remission (PR) and five induced or sustained complete remissions (CR). Five of these nine responders died during follow-up. The other patients still maintain CR with a current overall survival of 874-1424 days (median: 1150 days). In conclusion, despite the dismal clinical prognosis of children refractory to standard therapy, immunotherapy with Lymphomun resulted in a favourable clinical outcome in this cohort of refractory paediatric patients. PMID:25495919

  13. Evaluation of the sensitivity and specificity of six HIV combined p24 antigen and antibody assays

    Microsoft Academic Search

    Thoai Duong Ly; Syria Laperche; Catherine Brennan; Ana Vallari; Anne Ebel; Jeffrey Hunt; Lynn Martin; David Daghfal; Gerald Schochetman; Sushil Devare

    2004-01-01

    In this study, we evaluated the performance of six HIV combined p24 antigen and antibody (Ag\\/Ab) assays versus two third-generation anti-HIV antibody assays. The assays were evaluated using p24 antigen panel of 31 HIV-1 subtypes (n=124), 25 HIV-1 seroconversion panels (n=176), HIV-1 antibody positive samples including group M subtypes and group O (n=559), HIV-2 positive samples (n=110), and unselected HIV

  14. Offered: Offered: Position(s): Position(s)

    E-print Network

    New Hampshire, University of

    pharmaceutical companies. Adimab's yeast-based monoclonal antibody discovery platform functions like a synthetic packages comprised of competitive salary, bonus, and equity components, in addition to a world class

  15. Self-Assembled Antibody Multimers through Peptide Nucleic Acid Conjugation

    PubMed Central

    Kazane, Stephanie A.; Axup, Jun Y; Kim, Chan Hyuk; Ciobanu, Mihai; Wold, Erik D.; Barluenga, Sofia; Hutchins, Benjamin A.; Schultz, Peter G.; Winssinger, Nicolas; Smider, Vaughn V.

    2013-01-01

    With the recent clinical success of bispecific antibodies, a strategy to rapidly synthesize and evaluate bispecific or higher order multispecific molecules could facilitate the discovery of new therapeutic agents. Here we show that unnatural amino acids (UAAs) with orthogonal chemical reactivity can be used to generate site-specific antibody-oligonucleotide conjugates. These constructs can then be self-assembled into multimeric complexes with defined composition, valency and geometry. Using this approach, we generated potent bispecific antibodies that recruit cytotoxic T lymphocytes to Her2 and CD20 positive cancer cells, as well as multimeric antibody fragments with enhanced activity. This strategy should accelerate the synthesis and in vitro characterization of antibody constructs with unique specificities and molecular architectures. PMID:23210862

  16. Evaluation of anti-mutated citrullinated vimentin antibodies, anti-cyclic citrullinated Peptide antibodies and rheumatoid factor in omani patients with rheumatoid arthritis.

    PubMed

    Al-Shukaili, Ahmed; Al-Ghafri, Saif; Al-Marhoobi, Safia; Alkaabi, Juma

    2012-01-01

    Rheumatoid factor (RF) is currently used in the diagnosis of rheumatoid arthritis (RA). The discovery of anticitrullinated protein autoantibodies has led to the development of various new tests, such as anti-cyclic citrullinated peptide (anti-CCP) antibodies, and anti-mutated citrullinated vimentin (anti-MCV) antibodies, to diagnose RA. The aims of this study were to determine the sensitivity and specificity of anti-MCV antibodies in comparison with anti-CCP antibodies and RF in Omani Arab patients with RA and compare our findings with published values from different ethnic groups. The sensitivity of anti-MCV antibodies was 72% with 87% specificity. For anti-CCP antibodies the sensitivity was 52% and the specificity was 97%. The sensitivity of RF was 57% with 94% specificity. Anti-CCP antibodies have higher diagnostic specificity and positive predictive value than RF and anti-MCV antibodies. Anti-MCV antibodies have the highest sensitivity when compared to anti-CCP antibodies and RF. Anti-MCV antibodies do not appear to be very useful in the diagnosis of RA. However, long-term study is required to find out whether anti-MCV antibodies can be used as predictive test for incidence of RA. PMID:22934118

  17. Evaluation of Anti-Mutated Citrullinated Vimentin Antibodies, Anti-Cyclic Citrullinated Peptide Antibodies and Rheumatoid Factor in Omani Patients with Rheumatoid Arthritis

    PubMed Central

    Al-Shukaili, Ahmed; Al-Ghafri, Saif; Al-Marhoobi, Safia; Alkaabi, Juma

    2012-01-01

    Rheumatoid factor (RF) is currently used in the diagnosis of rheumatoid arthritis (RA). The discovery of anticitrullinated protein autoantibodies has led to the development of various new tests, such as anti-cyclic citrullinated peptide (anti-CCP) antibodies, and anti-mutated citrullinated vimentin (anti-MCV) antibodies, to diagnose RA. The aims of this study were to determine the sensitivity and specificity of anti-MCV antibodies in comparison with anti-CCP antibodies and RF in Omani Arab patients with RA and compare our findings with published values from different ethnic groups. The sensitivity of anti-MCV antibodies was 72% with 87% specificity. For anti-CCP antibodies the sensitivity was 52% and the specificity was 97%. The sensitivity of RF was 57% with 94% specificity. Anti-CCP antibodies have higher diagnostic specificity and positive predictive value than RF and anti-MCV antibodies. Anti-MCV antibodies have the highest sensitivity when compared to anti-CCP antibodies and RF. Anti-MCV antibodies do not appear to be very useful in the diagnosis of RA. However, long-term study is required to find out whether anti-MCV antibodies can be used as predictive test for incidence of RA. PMID:22934118

  18. Secondary Erythermalgia Associated with an Autoimmune Disorder of Undetermined Significance

    Microsoft Academic Search

    J. P. H. Drenth; J. J. M. Michiels; T. Van Joost; V. D. Vuzevski

    1995-01-01

    A 50-year-old female patient is described with an acquired, persisting and yet incurable erythermalgia featured by symmetric burning pain and red congestion of the extremities secondary to cutaneous vasculitis. A weakly positive antinuclear antibody titer and high titers of antibodies against gastric parietal mucosa cells pointed to an underlying but unclassifiable autoimmune disorder. It is concluded that histopathology of lesional

  19. Glycoproteomic Analysis of Antibodies*

    PubMed Central

    Zauner, Gerhild; Selman, Maurice H. J.; Bondt, Albert; Rombouts, Yoann; Blank, Dennis; Deelder, André M.; Wuhrer, Manfred

    2013-01-01

    Antibody glycosylation has been shown to change with various processes. This review presents mass spectrometric approaches for antibody glycosylation analysis at the level of released glycans, glycopeptides, and intact protein. With regard to IgG fragment crystallizable glycosylation, mass spectrometry has shown its potential for subclass-specific, high-throughput analysis. In contrast, because of the vast heterogeneity of peptide moieties, fragment antigen binding glycosylation analysis of polyclonal IgG relies entirely on glycan release. Next to IgG, IgA has gained some attention, and studies of its O- and N-glycosylation have revealed disease-associated glycosylation changes. Glycoproteomic analyses of IgM and IgE are lagging behind but should complete our picture of glycosylation's influence on antibody function. PMID:23325769

  20. Selection of recombinant antibodies from antibody gene libraries.

    PubMed

    Hust, Michael; Frenzel, André; Schirrmann, Thomas; Dübel, Stefan

    2014-01-01

    Antibodies are indispensable detection reagents for research and diagnostics and represent the biggest class of biological therapeutics on the market. In vitro antibody selection systems offer many advantages over animal-based technologies because the whole selection process is independent of the in vivo immune response. In the last two decades antibody phage display has evolved to the most robust and widely used method and has already yielded thousands of antibodies. The selection of binders by phage display is also referred to as "panning" and based on the specific molecular interaction of antibody phage with an immobilized antigen thus allowing the enrichment and isolation of antigen-specific monoclonal binders from very large antibody gene libraries. Here, we give detailed protocols for the selection of recombinant antibody fragments from antibody gene libraries in microtiter plates. PMID:24233787

  1. Enzyme-linked immunosorbent assay for measurement of cytomegalovirus glycoprotein B antibody in serum.

    PubMed

    Hackett, Daniel J; Zhang, Changpin; Stefanescu, Carla; Pass, Robert F

    2010-05-01

    Measurement of antibody to cytomegalovirus (CMV) glycoprotein B (gB) is valuable in the assessment of the antibody response to infection and to gB-containing vaccines. For this purpose, an enzyme-linked immunosorbent assay (ELISA) with a recombinant CMV gB molecule as the antigen was evaluated. Sera from 168 anti-CMV IgG-positive and 100 seronegative subjects were used to evaluate the anti-gB antibody assay. A cutoff optical density (OD) that would distinguish gB antibody-positive from -negative sera was established. Titers of antibody to gB determined by endpoint dilution were compared with those calculated using regression analysis. The run-to-run and interoperator reproducibilities of results were measured. The mean OD + 5 standard deviations from 50 anti-CMV IgG antibody-negative sera (0.2472) was used as the cutoff between anti-gB antibody-positive and -negative results. All sera from 100 anti-CMV IgG-seronegative subjects were negative for antibody to gB. All but 1 of 168 sera from seropositive subjects were positive for antibody to gB. Observed antibody levels based on titration to the endpoint were very similar to results calculated using linear regression. The run-to-run consistency of endpoints was excellent, with 38 runs from one operator and 48 runs from another all giving results within 1 dilution of the mean value for each of three anti-CMV IgG antibody-positive serum pools. The geometric mean titer of antibody to gB for 99 sera from seropositive blood donors was 1/10,937. This ELISA gives accurate and reproducible results for the relative quantity of anti-CMV gB IgG in serum over a wide range of antibody levels. PMID:20219875

  2. Oral lichen planus with antibodies to desmogleins 1 and 3.

    PubMed

    Kinjyo, Chihiro; Kaneko, Takahide; Korekawa, Ayumi; Rokunohe, Akiko; Aizu, Takayuki; Matsuzaki, Yasushi; Nakano, Hajime; Sawamura, Daisuke

    2015-01-01

    Lichen planus (LP) is a chronic inflammatory disorder involving the skin or mucous membranes. Previous studies have demonstrated that some LP patients showed positive enzyme-linked immunosorbent assay (ELISA) for desmoglein (DSG) antibodies. We report a case with intractable painful oral lesions. ELISA indices for DSG1 and 3 antibodies were increased by 49 and 36, respectively. Histopathological analysis revealed irregular acanthosis and band-like infiltration of lymphocytes at the dermal-epidermal interface. Direct immunofluorescence revealed negative deposits of immunoglobulin G and C3 in intracellular spaces of the epidermis. Indirect immunofluorescence of normal skin also did not detect any antibodies. Consequently, we made a final diagnosis of oral LP. The previous two LP cases with positive ELISA for DSG antibodies and our case manifested the erosive form, the most advanced oral LP. Therefore, it is a possibility that severe damage of keratinocytes may induce generation of DSG antibodies. However, negative results of immunofluorescence and no relation between disease severity and titers of antibodies make the possibility unlikely. We should measure titers of DSG antibodies in LP patients and accumulate data to establish a valid conclusion. PMID:25354571

  3. Sandwich antibody arrays using recombinant antibody-binding protein L.

    PubMed

    Seo, Jin-soo; Poulter, C Dale

    2014-06-10

    Antibody arrays are a useful for detecting antigens and other antibodies. This technique typically requires a uniform and well-defined orientation of antibodies attached to a surface for optimal performance. A uniform orientation can be achieved by modification of antibodies to include a single site for attachment. Thus, uniformly oriented antibody arrays require a bioengineered modification for the antibodies directly immobilization on the solid surface. In this study, we describe a "sandwich-type" antibody array where unmodified antibodies are oriented through binding with regioselectively immobilized recombinant antibody-binding protein L. Recombinant proL-CVIA bearing C-terminal CVIA motif is post-translationally modified with an alkyne group by protein farnesyltransferase (PFTase) at the cysteine residue in the CVIA sequence to give proL-CVIApf, which is covalently attached to an azido-modified glass slide by a Huisgen [3 + 2] cycloaddition reaction. Slides bearing antibodies bound to slides coated with regioselectively immobilized proL-CVIApf gave stronger fluorescence outputs and those where the antibody-binding protein was immobilized in random orientations on an epoxy-modified slide. Properly selected capture and detection antibodies did not cross-react with immobilized proL-CVIApf in sandwich arrays, and the proL-CVIApf slides can be used for multiple cycles of detected over a period of several months. PMID:24841983

  4. Radioimmunodetection of human melanoma tumor xenografts with human monoclonal antibodies.

    PubMed

    Gomibuchi, M; Saxton, R E; Lake, R R; Katano, M; Irie, R F

    1986-01-01

    We established a human IgM monoclonal antibody that defines a tumor-associated membrane antigen expressed on human melanoma cells. The antigen has been identified as the ganglioside GD2. In this paper, we describe the potential usefulness of the human monoclonal antibody for radioimaging. Nude mice bearing tumors derived from a human melanoma cell line were used as a model. Antibody activity was degradated significantly after labeling with 131I by the use of a modified chloramine-T method. After testing various concentrations, labeled antibody of a specific activity of 2.8 microCi/micrograms produced the best results. Balb/c nude mice bearing a GD2-positive M14 melanoma cell line were injected with 10-30 micrograms of labeled antibody, and its radiolocalization in different organs and in the whole body were evaluated. The best tumor image was obtained on Day 6. The labeled antibody uptake ratio between tumor and muscle was 9.2:1; the ratio between tumor and liver was 1.4:1. These studies represent the first report of experimental tumor imaging with human monoclonal antibody. Human monoclonals with probably prove to be superior reagents for tumor imaging in melanoma patients if the problem of antibody radiolysis is resolved. PMID:3771234

  5. Prevalence of Antibodies to Hepatitis C Virus in Hemodialysis Patients

    Microsoft Academic Search

    B. Kallinowski; L. Theilmann; K. Gmelin; K. Andrassy; D. Deppermann; B. Weinel; B. Kommerell

    1991-01-01

    Due to frequent parenteral contact with blood transfusions hemodialysis patients are prone to acquire hepatitis C virus (HCV) infection. To determine the role of HCV infection we investigated the prevalence of antibodies to HCV in 188 hemodialysis patients. The prevalence of antibodies to HCV was 7.4%. As compared to anti-HCV-negative patients, anti-HCV-positive patients had slightly elevated transaminases which were independent

  6. Antiphospholipid antibodies in patients with sensorineural hearing loss

    Microsoft Academic Search

    Edgar Bachor; Stephan Kremmer; Ernst Kreuzfelder; Klaus Jahnke; Said Seidahmadi

    2005-01-01

    Sensorineural hearing loss can be associated with autoimmune diseases and the presence of antiphospholipid antibodies. Sixty patients (mean age 47 years, range 18–76 years) with sudden sensorineural hearing loss were studied with audiograms, stapedial thresholds, otoacoustic emissions, positional and caloric testing. The serologic testing included antibodies against phosphatidylserine and ?2-glycoprotein. Additionally, a group of 34 patients (mean age 65 years, range 31–81 years) with

  7. Reshaping Antibody Diversity

    PubMed Central

    Wang, Feng; Ekiert, Damian C.; Ahmad, Insha; Yu, Wenli; Zhang, Yong; Bazirgan, Omar; Torkamani, Ali; Raudsepp, Terje; Mwangi, Waithaka; Criscitiello, Michael F.; Wilson, Ian A.; Schultz, Peter G.; Smider, Vaughn V.

    2014-01-01

    Summary Unlike humans or mice, some species have limited genome encoded combinatorial diversity potential, yet mount a robust antibody response. Cows are unusual in having exceptionally long CDR H3 loops and few V-regions, but the mechanism for creating diversity is not understood. Deep sequencing revealed that ultralong CDR H3s contain a remarkable complexity of cysteines, suggesting that disulfide-bonded mini-domains may arise during repertoire development. Indeed, crystal structures of two cow antibodies reveal that these CDR H3s form a very unusual architecture composed of a ?-strand “stalk” that supports a structurally diverse, disulfide-bonded, “knob” domain. Sequence analysis suggests that diversity arises from somatic hypermutation of an ultralong DH with a severe codon bias towards mutation to cysteine. These unusual antibodies can be elicited to recognize defined antigens through the knob domain. Thus, the bovine immune system produces an antibody repertoire composed of CDR H3s of unprecedented length that fold into a diversity of mini-domains generated through combinations of somatically generated disulfides. PMID:23746848

  8. Natural antibodies to glycans.

    PubMed

    Bovin, N V

    2013-07-01

    A wide variety of so-called natural antibodies (nAbs), i.e. immunoglobulins generated by B-1 cells, are directed to glycans. nAbs to glycans can be divided in three groups: 1) conservative nAbs, i.e. practically the same in all healthy donors with respect to their epitope specificity and level in blood; 2) allo-antibodies to blood group antigens; 3) plastic antibodies related to the first or the second group but discussed separately because their level changes considerably during diseases and some temporary conditions, in particular inflammation and pregnancy. Antibodies from the third group proved to be prospective markers of a number of diseases, whereas their unusual level (below or above the norm) is not necessarily the consequence of disease/state. Modern microarrays allowed the determination of the human repertoire, which proved to be unexpectedly broad. It was observed that the content of some nAbs reaches about 0.1% of total immunoglobulins. Immunoglobulins of M class dominate for most nAbs, constituting up to 80-90%. Their affinity (to a monovalent glycan, in KD terms) were found to be within the range 10(-4)-10(-6) M. Antibodies to Gal?1-3GlcNAc (Le(C)), 4-HSO3Gal?1-4GalNAc (4'-O-SuLN), Fuc?1-3GlcNAc, Fuc?1-4GlcNAc, GalNAc?1-3Gal (Adi), Gal?1-4Gal?1-4Glc (P(k)), Gal?1-4Gal?1-4GlcNAc (P1), GlcNAc?-terminated glycans, and hyaluronic acid should be noted among the nAbs revealed and studied during the last decade. At the same time, a kind of "taboo" is observed for a number of glycans: antibodies to Le(X) and Le(Y), and almost all gangliosides have not been observed in healthy persons. Many of the revealed nAbs were directed to constrained inner (core) part of glycan, directly adjoined to lipid of cell membrane or protein. The biological function of these nAbs remains unclear; for anti-core antibodies, a role of surveillance on appearance of aberrant, especially cancer, antigens is supposed. The first data related to oncodiagnostics based on quantitation of anti-glycan nAbs are reported. PMID:24010841

  9. Anti-neutrophil cytoplasmic antibody vasculitis presenting with bilateral renal vein thrombosis

    PubMed Central

    Robson, Michael Gregory

    2012-01-01

    We report a case of anti-neutrophil cytoplasmic antibody (ANCA)-associated necrotizing crescentic glomerulonephritis presenting with bilateral renal vein thrombosis and pulmonary emboli in a patient who also had a lupus anticoagulant and anti-cardiolipin antibodies. Although the link between venous thrombosis and ANCA vasculitis is well established, the coexistence of renal vein thrombosis is unusual. Furthermore, despite the positive ANCA, he was initially negative for antibodies to myeloperoxidase (MPO) and proteinase-3 (PR3), illustrating that a positive ANCA may be significant despite a negative test for antibodies to MPO and PR3.

  10. Transfer of maternal antibodies against avian influenza virus in mallards (Anas platyrhynchos).

    PubMed

    van Dijk, Jacintha G B; Mateman, A Christa; Klaassen, Marcel

    2014-01-01

    Maternal antibodies protect chicks from infection with pathogens early in life and may impact pathogen dynamics due to the alteration of the proportion of susceptible individuals in a population. We investigated the transfer of maternal antibodies against avian influenza virus (AIV) in a key AIV host species, the mallard (Anas platyrhynchos). Combining observations in both the field and in mallards kept in captivity, we connected maternal AIV antibody concentrations in eggs to (i) female body condition, (ii) female AIV antibody concentration, (iii) egg laying order, (iv) egg size and (v) embryo sex. We applied maternity analysis to the eggs collected in the field to account for intraspecific nest parasitism, which is reportedly high in Anseriformes, detecting parasitic eggs in one out of eight clutches. AIV antibody prevalence in free-living and captive females was respectively 48% and 56%, with 43% and 24% of the eggs receiving these antibodies maternally. In both field and captive study, maternal AIV antibody concentrations in egg yolk correlated positively with circulating AIV antibody concentrations in females. In the captive study, yolk AIV antibody concentrations correlated positively with egg laying order. Female body mass and egg size from the field and captive study, and embryos sex from the field study were not associated with maternal AIV antibody concentrations in eggs. Our study indicates that maternal AIV antibody transfer may potentially play an important role in shaping AIV infection dynamics in mallards. PMID:25386907

  11. Transfer of Maternal Antibodies against Avian Influenza Virus in Mallards (Anas platyrhynchos)

    PubMed Central

    van Dijk, Jacintha G. B.; Mateman, A. Christa; Klaassen, Marcel

    2014-01-01

    Maternal antibodies protect chicks from infection with pathogens early in life and may impact pathogen dynamics due to the alteration of the proportion of susceptible individuals in a population. We investigated the transfer of maternal antibodies against avian influenza virus (AIV) in a key AIV host species, the mallard (Anas platyrhynchos). Combining observations in both the field and in mallards kept in captivity, we connected maternal AIV antibody concentrations in eggs to (i) female body condition, (ii) female AIV antibody concentration, (iii) egg laying order, (iv) egg size and (v) embryo sex. We applied maternity analysis to the eggs collected in the field to account for intraspecific nest parasitism, which is reportedly high in Anseriformes, detecting parasitic eggs in one out of eight clutches. AIV antibody prevalence in free-living and captive females was respectively 48% and 56%, with 43% and 24% of the eggs receiving these antibodies maternally. In both field and captive study, maternal AIV antibody concentrations in egg yolk correlated positively with circulating AIV antibody concentrations in females. In the captive study, yolk AIV antibody concentrations correlated positively with egg laying order. Female body mass and egg size from the field and captive study, and embryos sex from the field study were not associated with maternal AIV antibody concentrations in eggs. Our study indicates that maternal AIV antibody transfer may potentially play an important role in shaping AIV infection dynamics in mallards. PMID:25386907

  12. Antibody mimetics: promising complementary agents to animal-sourced antibodies.

    PubMed

    Baloch, Abdul Rasheed; Baloch, Abdul Wahid; Sutton, Brian J; Zhang, Xiaoying

    2014-09-29

    Abstract Despite their wide use as therapeutic, diagnostic and detection agents, the limitations of polyclonal and monoclonal antibodies have inspired scientists to design the next generation biomedical agents, so-called antibody mimetics that offer many advantages over conventional antibodies. Antibody mimetics can be constructed by protein-directed evolution or fusion of complementarity-determining regions through intervening framework regions. Substantial progress in exploiting human, butterfly (Pieris brassicae) and bacterial systems to design and select mimetics using display technologies has been made in the past 10 years, and one of these mimetics [Kalbitor® (Dyax)] has made its way to market. Many challenges lie ahead to develop mimetics for various biomedical applications, especially those for which conventional antibodies are ineffective, and this review describes the current characteristics, construction and applications of antibody mimetics compared to animal-sourced antibodies. The possible limitations of mimetics and future perspectives are also discussed. PMID:25264572

  13. Subcutaneous Administration of Monoclonal Antibodies in Oncology

    PubMed Central

    Jackisch, C.; Müller, V.; Maintz, C.; Hell, S.; Ataseven, B.

    2014-01-01

    Treatment with monoclonal antibodies (mabs) has become an established component of oncological therapy. The monoclonal antibodies available for this purpose are mainly administered intravenously in individually adapted doses according to body weight over longer treatment times. For other chronic diseases such as, for example, diabetes mellitus, the subcutaneous administration of drugs is an established therapy option. For the subcutaneous administration of larger volumes as needed for mab solutions the extracellular matrix of the subcutaneous tissue represents a problem. The co-formulation with recombinant human hyaluronidase makes the relatively pain-free administration of larger fluid volumes and thus the subcutaneous administration of monoclonal antibodies possible, as illustrated by the development of a subcutaneous formulation of trastuzumab. This constitutes a less invasive, time-optimised and flexible form of administration for patients with HER2-positive breast cancer that, with its fixed dosing possibilities, contributes to therapeutic safety. The example of trastuzumab shows that the subcutaneous administration of monoclonal antibodies can simplify oncological long-term therapy not only for the patients but also for the medical personnel. PMID:25076790

  14. Site-specific antibody-drug conjugation through glycoengineering.

    PubMed

    Zhou, Qun; Stefano, James E; Manning, Charlene; Kyazike, Josephine; Chen, Bo; Gianolio, Diego A; Park, Anna; Busch, Michelle; Bird, Julie; Zheng, Xiaoyang; Simonds-Mannes, Helene; Kim, Jennifer; Gregory, Rick C; Miller, Robert J; Brondyk, William H; Dhal, Pradeep K; Pan, Clark Q

    2014-03-19

    Antibody-drug conjugates (ADCs) have been proven clinically to be more effective anti-cancer agents than native antibodies. However, the classical conjugation chemistries to prepare ADCs by targeting primary amines or hinge disulfides have a number of shortcomings including heterogeneous product profiles and linkage instability. We have developed a novel site-specific conjugation method by targeting the native glycosylation site on antibodies as an approach to address these limitations. The native glycans on Asn-297 of antibodies were enzymatically remodeled in vitro using galactosyl and sialyltransferases to introduce terminal sialic acids. Periodate oxidation of these sialic acids yielded aldehyde groups which were subsequently used to conjugate aminooxy functionalized cytotoxic agents via oxime ligation. The process has been successfully demonstrated with three antibodies including trastuzumab and two cytotoxic agents. Hydrophobic interaction chromatography and LC-MS analyses revealed the incorporation of ~1.6 cytotoxic agents per antibody molecule, approximating the number of sialic acid residues. These glyco-conjugated ADCs exhibited target-dependent antiproliferative activity toward antigen-positive tumor cells and significantly greater antitumor efficacy than naked antibody in a Her2-positive tumor xenograft model. These findings suggest that enzymatic remodeling combined with oxime ligation of the native glycans of antibodies offers an attractive approach to generate ADCs with well-defined product profiles. The site-specific conjugation approach presented here provides a viable alternative to other methods, which involve a need to either re-engineer the antibody sequence or develop a highly controlled chemical process to ensure reproducible drug loading. PMID:24533768

  15. Targeting antibodies to the cytoplasm

    PubMed Central

    Marschall, Andrea L J; Frenzel, André; Schirrmann, Thomas; Schüngel, Manuela

    2011-01-01

    A growing number of research consortia are now focused on generating antibodies and recombinant antibody fragments that target the human proteome. A particularly valuable application for these binding molecules would be their use inside a living cell, e.g., for imaging or functional intervention. Animal-derived antibodies must be brought into the cell through the membrane, whereas the availability of the antibody genes from phage display systems allows intracellular expression. Here, the various technologies to target intracellular proteins with antibodies are reviewed. PMID:21099369

  16. [Antibody therapy for Alzheimer's disease].

    PubMed

    Tabira, Takeshi; Matsumoto, Shin-Ei; Jin, Haifeng

    2011-11-01

    In order to avoid Abeta-induced autoimmune encephalitis, several monoclonal and polyclonal antibodies are in clinical trials. These are bapineuzumab, solanezumab, ponezumab, gantenerumab, BAN2401, gammaguard and octagam. Since each antibody has a different antigen epitope of Abeta, anti-amyloid activities are different. It is unknown which antibody is effective for Alzheimer disease, and we must wait for the result of clinical trials. Some patients who developed tissue amyloid plaque immuno-reactive (TAPIR) antibody showed slower decline after AN-1792 vaccination. We developed TAPIR-like monoclonal antibody, which was found to react with Abeta oligomers preferentially. PMID:22277519

  17. Therapeutic antibodies against cancer

    PubMed Central

    Adler, Mark J.; Dimitrov, Dimiter S.

    2012-01-01

    Antibody-based therapeutics against cancer are highly successful in clinic and currently enjoy unprecedented recognition of their potential; 13 monoclonal antibodies (mAbs) have been approved for clinical use in the European Union and in the United States (one, mylotarg, was withdrawn from market in 2010). Three of the mAbs (bevacizumab, rituximab, trastuzumab) are in the top six selling protein therapeutics with sales in 2010 of more than $5 bln each. Hundreds of mAbs including bispecific mAbs and multispecific fusion proteins, mAbs conjugated with small molecule drugs and mAbs with optimized pharmacokinetics are in clinical trials. However, challenges remain and it appears that deeper understanding of mechanisms is needed to overcome major problems including resistance to therapy, access to targets, complexity of biological systems and individual variations. PMID:22520975

  18. Antibody-mediated radiotherapy

    SciTech Connect

    Bloomer, W.D.; Lipsztein, R.; Dalton, J.F.

    1985-05-01

    Antibodies that react with antigens on the surface of tumor cells but not normal cells have great potential for cancer detection and therapy. If radiolabeled without loss of immunologic specificity, such antibodies may be able to deliver cytoxic amounts of radiation. Target- cell specificity and a high extraction coefficient are necessary with any radionuclide in order to minimize normal tissue irradiation. Tumor- cell-retention time and the rate of catabolized radionuclide will also influence ultimate applicability. Among the unanswered questions for choosing a radionuclide is the choice of particle emitter. Although classic beta emitters have been used in a number of clinical situations, they have not had a major impact on disease outcome except in diseases of the thyroid. Unfortunately, Auger emitters such as iodine 125 are cytotoxic only when localized within close proximity to the genome. On the other hand, alpha emitters such as astatine 211 eliminate the need for subcellular sequestration but not cell-specific localization. 34 references.

  19. Production of target-specific recombinant human polyclonal antibodies in mammalian cells.

    PubMed

    Wiberg, Finn C; Rasmussen, Søren K; Frandsen, Torben P; Rasmussen, Lone K; Tengbjerg, Kaja; Coljee, Vincent W; Sharon, Jacqueline; Yang, Chiou-Ying; Bregenholt, Søren; Nielsen, Lars S; Haurum, John S; Tolstrup, Anne B

    2006-06-01

    We describe the expression and consistent production of a first target-specific recombinant human polyclonal antibody. An anti-Rhesus D recombinant polyclonal antibody, Sym001, comprised of 25 unique human IgG1 antibodies, was produced by the novel Sympress expression technology. This strategy is based on site-specific integration of antibody genes in CHO cells, using the FRT/Flp-In recombinase system. This allows integration of the expression construct at the same genomic site in the host cells, thereby reducing genomic position effects. Different bioreactor batches of Sym001 displayed highly consistent manufacturing yield, antibody composition, binding potency, and functional activity. The results demonstrate that diverse recombinant human polyclonal antibody compositions can be reproducibly generated under conditions directly applicable to industrial manufacturing settings and present a first recombinant polyclonal antibody which could be used for treatment of hemolytic disease of the newborn and/or idiopathic thrombocytopenic purpura. PMID:16596663

  20. Antibodies for Immunohistochemistry

    Microsoft Academic Search

    Igor B. Buchwalow; Werner Böcker

    \\u000a The first use of the term Antikörper (the German word for antibody) occurred in a text by Paul Ehrlich (Fig. 1.1) in the conclusion of his article “Experimental Studies on Immunity,” published\\u000a in October 1891. Paul Ehrlich was born in 1854 in Strehlen (the German Province of Silesia, now in Poland). As a schoolboy\\u000a and student of medicine he was

  1. Antineutrophil cytoplasmic antibodies.

    PubMed

    Bosch, Xavier; Guilabert, Antonio; Font, Josep

    2006-07-29

    Much like other autoantibodies (eg, anti-double stranded DNA in systemic lupus erythematosus or antiglomerular basement membrane antibodies in Goodpasture's syndrome), antineutrophil cytoplasmic antibodies (ANCA) have provided doctors with a useful serological test to assist in diagnosis of small-vessel vasculitides, including Wegener's granulomatosis, microscopic polyangiitis, Churg-Strauss syndrome, and their localised forms (eg, pauci-immune necrotising and crescentic glomerulonephritis). 85-95% of patients with Wegener's granulomatosis, microscopic polyangiitis, and pauci-immune necrotising and crescentic glomerulonephritis have serum ANCA. ANCA directed to either proteinase 3 or myeloperoxidase are clinically relevant, yet the relevance of other ANCA remains unknown. Besides their diagnostic potential, ANCA might be valuable in disease monitoring. In addition, data seem to confirm the long-disputed pathogenic role of these antibodies. Present treatments for ANCA-associated vasculitis are not free from side-effects and as many as 50% of patients relapse within 5 years. Accurate understanding of the key pathogenic points of ANCA-associated vasculitis can undoubtedly provide a more rational therapeutic approach. PMID:16876669

  2. Evaluation of antibody frequency against HBV, HCV and HTLV-1

    PubMed Central

    Tahaei, Seyed Mohammad Ebrahim; Fatemi, Seyed Reza; Azimzadeh, Pedram; Mirsattari, Dariush; Sanati, Azar; Sharifian, Afsaneh

    2012-01-01

    Aim This study was designed to evaluate the frequency of antibody against these viruses in individuals attending the endoscopy ward of Taleghani hospital Tehran, Iran. Background Blood-borne viruses such as hepatitis B and hepatitis C virus and HTLV-1 virus are among the world’s public health problems. Hepatitis viruses cause liver problems and HTLV-1 infection can lead to adult T-Cell lymphoma (ATL). Patients and methods Blood samples of 219 individuals attending the endoscopy ward of Taleghani hospital between years 2009-2011 were collected. A questionnaire containing demographic data was completed for each subject. Blood samples were tested for antibody against HTLV-1, HCV and HBc by ELISA (Dia.pro Italy). In case of positive results for anti-HBc, samples were also tested for HBs Ag antigen. Results Ninety two subjects were male and 127 were female. Mean age of the population was 39.87 ± 16.47. None of the subjects had anti-HCV antibody, while 4 of them had anti-HTLV-1 antibody and 26 anti-HBc antibody; which only two of these individuals had HBs Antibody. Conclusion The results of this study show that frequency of anti-HCV and anti-HTLV-1 antibodies are very low, while the frequency of anti-HBc was higher in the population. Since HTLV-1 is the causative agent of a type of blood cancer, it seems that screening of donated bloods in this region should be considered. PMID:24834218

  3. Anti-Plasminogen Antibodies Compromise Fibrinolysis and Associate with Renal Histology in ANCA-Associated Vasculitis

    PubMed Central

    Berden, Annelies E.; Nolan, Sarah L.; Morris, Hannah L.; Bertina, Rogier M.; Erasmus, Dianhdra D.; Hagen, E. Christiaan; Hayes, Donal P.; van Tilburg, Nico H.; Bruijn, Jan A.; Savage, Caroline O.S.; Bajema, Ingeborg M.

    2010-01-01

    Antibodies recognizing plasminogen, a key component of the fibrinolytic system, associate with venous thrombotic events in PR3-ANCA vasculitis. Here, we investigated the prevalence and function of anti-plasminogen antibodies in independent UK and Dutch cohorts of patients with ANCA-associated vasculitis (AAV). We screened Ig isolated from patients (AAV-IgG) and healthy controls by ELISA. Eighteen of 74 (24%) UK and 10/38 (26%) Dutch patients with AAV had anti-plasminogen antibodies compared with 0/50 and 1/61 (2%) of controls. We detected anti-plasminogen antibodies in both PR3-ANCA– and MPO-ANCA–positive patients. In addition, we identified anti-tissue plasminogen activator (tPA) antibodies in 13/74 (18%) patients, and these antibodies were more common among patients with anti-plasminogen antibodies (P = 0.011). Eighteen of 74 AAV-IgG (but no control IgG) retarded fibrinolysis in vitro, and this associated with anti-plasminogen and/or anti-tPA antibody positivity. Only 4/18 AAV-IgG retarded fibrinolysis without harboring these antibodies; dual-positive samples retarded fibrinolysis to the greatest extent. Patients with anti-plasminogen antibodies had significantly higher percentages of glomeruli with fibrinoid necrosis (P < 0.05) and cellular crescents (P < 0.001) and had more severely reduced renal function than patients without these antibodies. In conclusion, anti-plasminogen and anti-tPA antibodies occur in AAV and associate with functional inhibition of fibrinolysis in vitro. Seropositivity for anti-plasminogen antibodies correlates with hallmark renal histologic lesions and reduced renal function. Conceivably, therapies that enhance fibrinolysis might benefit a subset of AAV patients. PMID:20847144

  4. Microbials for the production of monoclonal antibodies and antibody fragments

    PubMed Central

    Spadiut, Oliver; Capone, Simona; Krainer, Florian; Glieder, Anton; Herwig, Christoph

    2014-01-01

    Monoclonal antibodies (mAbs) and antibody fragments represent the most important biopharmaceutical products today. Because full length antibodies are glycosylated, mammalian cells, which allow human-like N-glycosylation, are currently used for their production. However, mammalian cells have several drawbacks when it comes to bioprocessing and scale-up, resulting in long processing times and elevated costs. By contrast, antibody fragments, that are not glycosylated but still exhibit antigen binding properties, can be produced in microbial organisms, which are easy to manipulate and cultivate. In this review, we summarize recent advances in the expression systems, strain engineering, and production processes for the three main microbials used in antibody and antibody fragment production, namely Saccharomyces cerevisiae, Pichia pastoris, and Escherichia coli. PMID:24183828

  5. Detection of immunoglobulin M antibodies to Treponema pallidum in a modified enzyme-linked immunosorbent assay

    Microsoft Academic Search

    F. Mtiller; M. Moskophidis; H.-L. Borkhardt

    1987-01-01

    The indirect enzyme-linked immunosorbent assay (ELISA) for detection of immunoglobulin M (IgM) antibodies toTreponema pallidum in sera of syphilitic patients is complicated by false positive reactions due to the interference of IgM rheumatoid factor (IgM-RF) activity and the presence of treponemal IgG antibodies. Another source of error producing false negative results is the competition between treponemal IgG and IgM antibodies

  6. A SURVEY OF HANTAVIRUS ANTIBODY IN SMALL-MAMMAL POPULATIONS IN SELECTED UNITED STATES NATIONAL PARKS

    Microsoft Academic Search

    JAMES N. MILLS; JERRY M. JOHNSON; THOMAS G. KSIAZEK; BARBARA A. ELLIS; PIERRE E. ROLLIN; TERRY L. YATES; MICHAEL O. MANN; MARK R. JOHNSON; MARIEL L. CAMPBELL; JENNIFER MIYASHIRO; MICHAEL PATRICK; MICHAEL ZYZAK; DAVE LAVENDER; MARK G. NOVAK; KARINA SCHMIDT; C. J. PETERS; JAMES E. CHILDS

    Hantavirus activity in 39 National Parks in the eastern and central United States was surveyed by testing 1,815 small mammals of 38 species for antibody reactive to Sin Nombre virus. Antibody-positive rodents were found throughout the area sampled, and in most biotic communities. Antibody was detected in 7% of 647 deer mice (Peromyscus maniculatus ), 2% of 590 white-footed mice

  7. Detection of Anti-Yellow Fever Virus Immunoglobulin M Antibodies at 3–4 Years Following Yellow Fever Vaccination

    PubMed Central

    Gibney, Katherine B.; Edupuganti, Srilatha; Panella, Amanda J.; Kosoy, Olga I.; Delorey, Mark J.; Lanciotti, Robert S.; Mulligan, Mark J.; Fischer, Marc; Staples, J. Erin

    2012-01-01

    The duration of anti-yellow fever (YF) virus immunoglobulin M (IgM) antibodies following YF vaccination is unknown, making it difficult to interpret positive IgM antibody results in previously vaccinated travelers. We evaluated the frequency and predictors of YF IgM antibody positivity 3–4 years following YF vaccination. Twenty-nine (73%) of 40 participants had YF IgM antibodies 3–4 years postvaccination. No demographic or exposure variables were predictive of YF IgM positivity. However, persons who were YF IgM positive at 3–4 years postvaccination had earlier onset viremia and higher neutralizing antibody geometric mean titers at 1 month and 3–4 years postvaccination compared with persons who were YF IgM negative. Detection of YF IgM antibodies several years postvaccination might reflect remote YF vaccination rather than recent YF vaccination or YF virus infection. PMID:23109371

  8. Immunofluorescent antibody test for diagnosis of gonorrhoea.

    PubMed Central

    Caloenescu, M; Clecner, B; Petrow, S; Kasatiya, S S

    1975-01-01

    An indirect fluorescent antibody test was evaluated in 198 cases of a high-risk group with a culture prevalence of 37.3% and in 426 cases of a low-risk group with a culture prevalence of 1.16%. A sensitivity of 77.1% in the culture-positive patients with uncomplicated gonorrhoea, and a specificity of 88.7% in the culture- and history-negative cases, was obtained in the high-risk group. In this group, the sera from 88.8% of the patients with culture-proven gonorrhoea became positive in an indirect fluorescent antibody test within 3 weeks of last sexual contact. In the low-risk group, for which the sensitivity could not be determined due to various reasons, a specificity of 95.8% was obtained. Complement fixation test was positive in sera of only 17.6% of the culture-positive cases of the high-risk group. PMID:809468

  9. Leucine-rich glioma-inactivated protein 1 antibody encephalitis

    PubMed Central

    Mayasi, Yunis; Takhtani, Deepak

    2014-01-01

    Objective: To describe a case of leucine-rich glioma-inactivated protein 1 (LGI1) antibody–associated encephalitis. Methods: The clinical and ancillary data and brain MRIs were gathered retrospectively by chart review. Relevant literature on similar cases was also reviewed. Results: The diagnosis of LGI1 antibody–associated autoimmune encephalitis was based on the typical clinical presentation of seizures, psychiatric symptoms, and memory loss as well as negative diagnostic testing for cancer; the diagnosis was confirmed by positive LGI1 antibody. The patient responded favorably to treatment with IV immunoglobulin and continues to do well. Conclusion: LGI1 antibody–associated encephalitis has increasingly been recognized as a primary autoimmune disorder with good prognosis and response to treatment. PMID:25520958

  10. Probable C4d-negative accelerated acute antibody-mediated rejection due to non-HLA antibodies.

    PubMed

    Niikura, Takahito; Yamamoto, Izumi; Nakada, Yasuyuki; Kamejima, Sahoko; Katsumata, Haruki; Yamakawa, Takafumi; Furuya, Maiko; Mafune, Aki; Kobayashi, Akimitsu; Tanno, Yudo; Miki, Jun; Yamada, Hiroki; Ohkido, Ichiro; Tsuboi, Nobuo; Yamamoto, Hiroyasu; Yokoo, Takashi

    2015-07-01

    We report a case of probable C4d-negative accelerated acute antibody-mediated rejection due to non-HLA antibodies. A 44 year-old male was admitted to our hospital for a kidney transplant. The donor, his wife, was an ABO minor mismatch (blood type O to A) and had Gitelman syndrome. Graft function was delayed; his serum creatinine level was 10.1?mg/dL at 3 days after transplantation. Open biopsy was performed immediately; no venous thrombosis was observed during surgery. Histology revealed moderate peritubular capillaritis and mild glomerulitis without C4d immunoreactivity. Flow cytometric crossmatching was positive, but no panel-reactive antibodies against HLA or donor-specific antibodies (DSAbs) to major histocompatibility complex class I-related chain A (MICA) were detected. Taken together, we diagnosed him with probable C4d-negative accelerated antibody-mediated rejection due to non-HLA, non-MICA antibodies, the patient was treated with steroid pulse therapy (methylprednisolone 500?mg/day for 3 days), plasma exchange, intravenous immunoglobulin (40?g/body), and rituximab (200?mg/body) were performed. Biopsy at 58 days after transplantation, at which time S-Cr levels were 1.56?mg/dL, found no evidence of rejection. This case, presented with a review of relevant literature, demonstrates that probable C4d-negative accelerated acute AMR can result from non-HLA antibodies. PMID:26031592

  11. Offered: Offered: Position(s): Position(s)

    E-print Network

    New Hampshire, University of

    training to introduce the basics of a career in the staffing industry -Assigned mentors to guide youCompany: Industry: Website: Majors: Offered: Offered: Position(s): Position(s): Description IT staffing and Global Services for 82% of the Fortune 500. Ranked #1 in the industry by IT Services Business

  12. Offered: Offered: Position(s): Position(s)

    E-print Network

    New Hampshire, University of

    overall impact to company?s profitability ?Run monthly reports supporting Financial Analysts includingCompany: Industry: Website: Majors: Offered: Offered: Position(s): Position(s): Description and creative companies making premium chocolate, with six production sites in Europe, two in the USA

  13. Interference of endogenous lactoperoxidase antibodies in a solid-phase immunosorbent radioassay for antibodies to protein hormones

    SciTech Connect

    Ericsson, U.B.; Larsson, I.

    1984-11-01

    Interference of endogenous antibodies to lactoperoxidase (EC 1.11.1.7) has been demonstrated in a solid-phase immunosorbent radioassay for the detection of antibodies to protein hormones. On enzymic iodination with lactoperoxidase, a little of the /sup 125/I is incorporated in the enzyme itself by self-iodination. Because the /sup 125/I-labeled lactoperoxidase was not removed from the tracer by chromatography on Sephadex G-50, G-100, and G-200 columns, endogenous antibodies to the enzyme, which were present in 25 of 28 sera from apparently healthy individuals, were detected in conjunction with the measurement of antibodies to growth hormone (somatotropin), prolactin, and thyroglobulin, thus causing false-positive results. Contaminating radioactive lactoperoxidase can be removed by adsorption chromatography on cellulose or by liquid chromatography, and can be avoided by performing the iodination with immobilized lactoperoxidase or with Chloramine-T.

  14. Off-rate screening for selection of high-affinity anti-drug antibodies.

    PubMed

    Ylera, Francisco; Harth, Stefan; Waldherr, Dirk; Frisch, Christian; Knappik, Achim

    2013-10-15

    The rapidly increasing number of therapeutic antibodies in clinical development and on the market requires corresponding detection reagents for monitoring the concentration of these drugs in patient samples and as positive controls for measurement of anti-drug antibodies. Phage display of large recombinant antibody libraries has been shown to enable the rapid development of fully human anti-idiotypic antibodies binding specifically to antibody drugs, since the in vitro panning approach allows for incorporation of suitable blockers to drive selection toward the paratope of the drug. A typical bottleneck in antibody generation projects is ranking of the many candidates obtained after panning on the basis of antibody binding strength. Ideally, such method will work without prior labeling of antigens and with crude bacterial lysates. We developed an off-rate screening method of crude Escherichia coli lysates containing monovalent Fab fragments obtained after phage display of the HuCAL PLATINUM® antibody library. We used the antibody drugs trastuzumab and cetuximab as antigen examples. Using the Octet® RED384 label-free sensor instrument we show that antibody off rates can be reliably determined in crude bacterial lysates with high throughput. We also demonstrate that the method can be applied to screening for high-affinity antibodies typically obtained after affinity maturation. PMID:23906643

  15. Antibody Production by Single Cells

    Microsoft Academic Search

    G. J. V. Nossal; Joshua Lederberg

    1958-01-01

    FAGREUS1 and others2,3 have shown that certain tissues from pre-sensitized animals can form antibody in vitro. This communication describes a technique whereby antibody production by single cells isolated in microdroplets can be detected. The technique is based on specific immobilization of Salmonella serotypes by anti-flagellar antibody. It was observed that single cells from a rat, simultaneously stimulated with two antigens,

  16. Human antibodies from transgenic animals

    Microsoft Academic Search

    Nils Lonberg

    2005-01-01

    Laboratory mice provide a ready source of diverse, high-affinity and high-specificity monoclonal antibodies (mAbs). However, development of rodent antibodies as therapeutic agents has been impaired by the inherent immunogenicity of these molecules. One technology that has been explored to generate low immunogenicity mAbs for in vivo therapy involves the use of transgenic mice expressing repertoires of human antibody gene sequences.

  17. Function-first antibody discovery

    PubMed Central

    Frendéus, Björn

    2013-01-01

    Therapeutic antibodies may mediate antineoplastic effects by altering the biological functions of their target, by directly stimulating the demise of cancer cells or by activating antibody-dependent immune effector mechanisms. We have recently provided in vivo proof-of-concept for a “function-first” target and drug discovery platform in which antibodies against a multitude of tumor-associated antigens are screened for biological effects in a target-unbiased manner. PMID:24083074

  18. Recovery and Purification of Antibody

    Microsoft Academic Search

    XueJun Han; Arthur Hewig; Ganesh Vedantham

    \\u000a Monoclonal antibody drugs have become a large portion of protein therapeutics and many antibody molecules are being evaluated\\u000a at various stages of clinical trials in the biopharmaceutical industry. This review article summarizes the state of the art\\u000a antibody purification techniques. The main focus is chromatographic techniques that include protein A, ion exchange and HIC.\\u000a For each technique, the mechanisms of

  19. A Monoclonal Antibody to Saxitoxin

    Microsoft Academic Search

    Rüdiger Hack; Volker Renz; Erwin Märtlbauer; Gerhard Terplan

    1990-01-01

    After immunization of six BALB\\/c mice with saxitoxin (STX) coupled to keyhole limpet hemocyanin only one mouse developed serum antibodies specific to STX. By hybridizing the splenocytes of this mouse with X63?Ag8.653 myeloma cells one hybridoma line secreting antibodies to STX was produced. The antibody was designated 5C2 and proved to be IgM. In an indirect enzyme immunoassay using STX

  20. Screening Autoimmune Anti-neuronal Antibodies in Pediatric Patients with Suspected Autoimmune Encephalitis

    PubMed Central

    Kim, Soo Yeon; Choi, Sun Ah; Ryu, Hye Won; Kim, Hunmin; Lim, Byung Chan; Hwang, Hee; Chae, Jong-Hee; Choi, Jieun; Kim, Ki Joong; Hwang, Yong Seung; Lee, Soon-Tae; Chu, Kon; Lee, Sang Kun

    2014-01-01

    Background and Purpose: The aim of this study was to identify and describe the pediatric autoimmune encephalitis cases positive for anti-neuronal antibody tests. Methods: Screening of six anti-neuronal antibodies in 23 children with suspected autoimmune encephalitis was performed by cell-based indirect immunofluorescence test with patients’ serum or cerebrospinal fluid. Results: Among the 23 cases enrolled here, eight patients (35%) were positive for the anti-N-methyl-d-aspartate (NMDA) receptor antibody and one patient (4%) was positive for the anti-contactin-associated protein-like 2 (CASPR2) antibody. In the anti-NMDA receptor antibody-positive group, seizure and movement disorders were the most prominent features and were present in all patients. A tumor was present in only one patient. Three patients with infant- and toddler-onset disease did not exhibit a classic multistage illness. In addition to seizure and dyskinesia, aphasia or mutism without severe consciousness impairment was present in all three patients. These atypical clinical presentations may suggest different pathomechanism of anti-NMDA receptor encephalitis among these age groups. The patient who was positive for the anti-CASPR2 antibody was an 8-year-old girl who presented with fever, encephalopathy, and seizure. Neuromyotonia or other dyskinesia was not present. Conclusions: Eight anti-NMDA receptor antibody positive patients and one CASPR2 positive patient were identified from the screening of six anti-neuronal antibodies in pediatric patients suspected with autoimmune encephalitis. Developmental regression specifically for language skills was suggested as one of the atypical clinical features in infants and toddler onset anti-NMDA receptor antibody positive patients. PMID:25625089

  1. "Cytoplasmic" islet cell antibodies. Evidence that the target antigen is a sialoglycoconjugate.

    PubMed

    Nayak, R C; Omar, M A; Rabizadeh, A; Srikanta, S; Eisenbarth, G S

    1985-06-01

    We have biochemically treated (periodate, borohydride, neuraminidase, organic solvents) frozen sections of human pancreas and studied the reactivity of islet-cell-antibody-positive human sera and monoclonal antibodies. The autoantigen of pancreatic sections has the properties of sialic acid containing glycolipid. PMID:2989050

  2. A longitudinal study on avian polyomavirus-specific antibodies in captive Spix's macaws (Cyanopsitta spixii).

    PubMed

    Deb, Amrita; Foldenauer, Ulrike; Borjal, Raffy Jim; Streich, W Jürgen; Lüken, Caroline; Johne, Reimar; Müller, Hermann; Hammer, Sven

    2010-09-01

    Avian polyomavirus (APV) causes a range of disease syndromes in psittacine birds, from acute fatal disease to subclinical infections, depending on age, species, and other unidentified risk factors. To determine the prevalence of APV-specific antibodies in a captive population of Spix's macaws (Cyanopsitta spixii) in Quatar, 54 birds were tested by blocking enzyme-linked immunosorbent assay. A prevalence of 48.1% for APV antibodies, which indicates viral exposure, was found. Of 36 Spix's macaws that were serially tested over a period of 4 years, 50.0% were consistently positive, 36.1% were consistently negative, 5.5% had permanently declining antibody levels, and 2.8% showed variable results. By using polymerase chain reaction testing on whole blood samples, an apparent viremia was detected in 1 of 44 birds (2.3%), although contamination provides a likely explanation for this isolated positive result in a hand-reared chick. The white blood cell count was significantly higher in antibody-positive birds compared with antibody-negative birds (P < .05). Because antibody-positive and antibody-negative birds were housed together without a change in their respective antibody status, transmission of APV within the adult breeding population appeared to be a rare event. PMID:21046939

  3. A MORBILLIVIRUS ANTIBODY SURVEY OF ATLANTIC WALRUS, NARWHAL AND BELUGA IN CANADA

    Microsoft Academic Search

    Ole Nielsen; Robert E. A. Stewart; Lena Measures; Padraig Duignan; Carol House

    2000-01-01

    A longitudinal serologic survey was conducted for morbillivirus antibodies in Atlantic walruses (Odobenus rosmarus rosmarus), narwhal (Monodon monoceros), and beluga (Delphi- napterus leucas) from the Northwest Territories, Nunavut and the St. Lawrence estuary (Canada). Sixty-five of 131 (50%) walruses sampled between 1984 and 1993 had detectable morbillivirus neutralizing antibodies. Positive walrus were identified from four of five Arctic sampling sites,

  4. Automated Instrument for the Fluorescent Treponemal Antibody-Absorption Test and Other Immunofluorescence Tests

    PubMed Central

    Binnings, Gerald F.; Riley, Mel J.; Roberts, Merritt E.; Barnes, Richard; Pringle, Thomas C.

    1969-01-01

    An automated diagnostic test instrument and its development program are described. The instrument automates the fluorescent treponemal antibody-absorption test for syphilis to the extent that only 4 hr of technician time is required to conduct approximately 200 tests daily. Evaluation to date suggests its efficacy. In addition, preliminary studies indicate the feasibility of detecting antibodies to Toxoplasma gondii, Plasmodium malariae, and nucleoprotein (antinuclear factor). The instrument would seem to have broad application for routine and research immunofluorescence testing. Two elements comprise the instrument: a slide processor and a microscope attachment. The slide processor is an electro-pneumatically actuated device which automatically feeds special laboratory slides, on which antigen or other reagents are prefixed, through a series of operations which provide reagent application, incubation, washing, drying, and stacking of the finished slides for readout. The instrument provides flexibility in that incubation time and temperature as well as point, sequence, and duration of reagent application can be varied to accommodate a variety of immunofluorescence techniques. The microscope attachment can be fitted to all conventional dark-field fluorescence microscopes and makes possible the reading of three to six slides per minute. The reacted slides from the processor are injected sequentially onto the stage of the microscope by movement of a lever. As injected, slides are automatically in visual focus; fine focus is occasionally required. Scanning of the reacted field is accomplished by means of the normal microscope controls. A buffered glycerol coupling is maintained between the darkfield condenser substage lens and the slide cover glass by means of a pushbutton-actuated feed system. Images PMID:4905605

  5. Islet Cell Antibodies Represent Autoimmune Response Against Several Antigens

    PubMed Central

    Månsson, Lisa; Törn, Carina

    2001-01-01

    To study the antigens involved in the islet cell antibody (ICA) reaction we selected 30 patient serum samples (ten in each group) positive for ICA and one other additional autoantibody, such as glutamic acid decarboxylase antibodies (GADA), thyrosine phosphatase antibodies (IA-2A) or insulin autoantibodies (IAA). The serum samples were incubated with the specific antigen (GAD65, IA-2 or insulin) and the ICA analysis and the corresponding immunoprecipitation assay were performed before and after the absorption. We could then demonstrate that specific autoantibodies against GAD65 and IA-2 could be absorbed with the corresponding antigen, since ten GADA positive and six IA-2A samples turned completely negative. However, the ICA reaction after absorption with GADA, IA-2A and insulin was still present, although at significantly lower levels. The results strongly indicate that the ICA reaction represents simultaneous autoimmunity against several other antigens beside GAD65, IA-2 and insulin. PMID:12369720

  6. High-content Analysis of Antibody Phage-display Library Selection Outputs Identifies Tumor Selective Macropinocytosis-dependent Rapidly Internalizing Antibodies*

    PubMed Central

    Ha, Kevin D.; Bidlingmaier, Scott M.; Zhang, Yafeng; Su, Yang; Liu, Bin

    2014-01-01

    Many forms of antibody-based targeted therapeutics, including antibody drug conjugates, utilize the internalizing function of the targeting antibody to gain intracellular entry into tumor cells. Ideal antibodies for developing such therapeutics should be capable of both tumor-selective binding and efficient endocytosis. The macropinocytosis pathway is capable of both rapid and bulk endocytosis, and recent studies have demonstrated that it is selectively up-regulated by cancer cells. We hypothesize that receptor-dependent macropinocytosis can be achieved using tumor-targeting antibodies that internalize via the macropinocytosis pathway, improving potency and selectivity of the antibody-based targeted therapeutic. Although phage antibody display libraries have been utilized to find antibodies that bind and internalize to target cells, no methods have been described to screen for antibodies that internalize specifically via macropinocytosis. We hereby describe a novel screening strategy to identify phage antibodies that bind and rapidly enter tumor cells via macropinocytosis. We utilized an automated microscopic imaging-based, High Content Analysis platform to identify novel internalizing phage antibodies that colocalize with macropinocytic markers from antibody libraries that we have generated previously by laser capture microdissection-based selection, which are enriched for internalizing antibodies binding to tumor cells in situ residing in their tissue microenvironment (Ruan, W., Sassoon, A., An, F., Simko, J. P., and Liu, B. (2006) Identification of clinically significant tumor antigens by selecting phage antibody library on tumor cells in situ using laser capture microdissection. Mol. Cell. Proteomics. 5, 2364–2373). Full-length human IgG molecules derived from macropinocytosing phage antibodies retained the ability to internalize via macropinocytosis, validating our screening strategy. The target antigen for a cross-species binding antibody with a highly active macropinocytosis activity was identified as ephrin type-A receptor 2. Antibody-toxin conjugates created using this macropinocytosing IgG were capable of potent and receptor-dependent killing of a panel of EphA2-positive tumor cell lines in vitro. These studies identify novel methods to screen for and validate antibodies capable of receptor-dependent macropinocytosis, allowing further exploration of this highly efficient and tumor-selective internalization pathway for targeted therapy development. PMID:25149096

  7. Anti-flavin antibodies.

    PubMed

    Barber, M J; Eichler, D C; Solomonson, L P; Ackrell, B A

    1987-02-15

    Antibodies were elicited to FAD by using the hapten N-6-(6-aminohexyl)-FAD conjugated to the immunogenic carrier protein bovine serum albumin. Cross-reactivity was determined by Ouchterlony double-diffusion analysis with N-6-(6-aminohexyl)-FAD coupled to rabbit serum albumin. Anti-FAD IgG was partially purified by (NH4)2SO4 precipitation followed by DEAE-cellulose/CM-cellulose and bovine serum albumin-agarose chromatography. The partially purified anti-FAD IgG fraction failed to inhibit the catalytic activities of the flavin-containing enzymes nitrate reductase, xanthine oxidase and succinate dehydrogenase, whereas enzyme activity could be inhibited by addition of antibodies elicited against the native proteins. However, the partially purified anti-FAD IgG fraction could be used as a highly sensitive and specific probe to detect proteins containing only covalently bound flavin, such as succinate dehydrogenase, p-cresol methylhydroxylase and monoamine oxidase, by immuno-blotting techniques. Detection limits were estimated to be of the order of femtomolar concentrations of FAD with increased sensitivity for the 8 alpha-N(3)-histidyl linkage compared with 8 alpha-O-tyrosyl substitution. PMID:3109386

  8. STUDIES ON ANTIBODY PRODUCTION

    PubMed Central

    Ambrose, Charles T.; Coons, Albert H.

    1963-01-01

    When lymph node fragments from previously immunized rabbits were stimulated in vitro to produce a secondary response, the continuous presence of 50 µg/ml (0.15 mM) of chloramphenicol in the medium during the entire incubation period of 15 to 21 days produced nearly complete suppression of the response. Concentrations as low as 5 µg/ml (0.015 mM) produced approximately 80 per cent suppression of the response. When 50 µg/ml of chloramphenicol was present during only the first 6 days of culture, the secondary response was reduced 90 per cent. When it was absent for the first 6 days but present for the next 9 to 15 days, the response was reduced only 40 per cent. Since over 95 per cent of the antibody of the secondary response in most experiments appeared in the medium after the 6th day, chloramphenicol apparently inhibits antibody production by interfering with some early phase of the response. It is suggested that this interference involves messenger RNA and that animal cells have appeared resistant to this drug only because their complement of messenger RNA present when the drug has been added is stable over the short periods during which protein synthesis has usually been studied. PMID:14012518

  9. Lung adenocarcinoma and antiphospholipid antibodies.

    PubMed

    de Meis, Ernesto; Monteiro, Robson Q; Levy, Roger A

    2009-05-01

    Thrombosis is a frequent finding in cancer patients, being referred to as a poor prognostic factor. The mechanisms underlying the thrombophilic state in malignancy are not well elucidated but involve a complex interaction between tumor and host cells as well as the hemostatic system. A number of studies have demonstrated the presence of antiphospholipid antibodies (aPL) in cancer patients, suggesting a potential role in tumor-associated thrombosis. A prospective analysis has been performed in a group of lung adenocarcinoma patients in respect to the presence of aPL and thrombotic manifestations. Lupus anticoagulant (LAC) was identified in 61 out of 105 patients and it correlated highly with thrombosis (22/61, LAC positive vs 2/44, LAC negative RR=7.93; p<0.001). On the other hand, patients that displayed IgM anti-beta2-glycoprotein I (abeta2GPI) (22/80) showed an unexpected decrease in thrombosis risk (2/22, with IgM abeta2GPI vs 18/58, without IgM abeta2GPI RR=0.29; p=0.04). Considerations on the mechanisms that link cancer, thrombosis and aPL are discussed in this article. PMID:19185619

  10. Seropositivity of Dengue Antibodies during Pregnancy

    PubMed Central

    Mohamed Ismail, Nor Azlin; Wan Abd Rahim, Wan Elly Rushima; Salleh, Sharifah Azura; Neoh, Hui-Min; Jamal, Rahman; Jamil, Muhammad Abdul

    2014-01-01

    Purpose. Malaysia a dengue endemic country with dengue infections in pregnancy on the rise. The present study was aimed at determining dengue seroprevalence (IgG or IgM) during pregnancy and its neonatal transmission in dengue seropositive women. Methods. Maternal with paired cord blood samples were tested for dengue antibodies (IgG and IgM) using an enzyme-linked immunosorbent assay (ELISA). Maternal age, parity, occupation, ethnic group, and gestational age were recorded. Data on neonatal Apgar score and admissions to the Neonatal Intensive Care Unit (NICU) were analyzed. Results. Out of 358 women recruited, about 128 (35.8%) patients were seropositive. Twelve patients (3.4%) had recent infections (IgM positive) and another 116 women (32.4%) were with past infections (IgG positive). All babies born to seropositive mothers had positive IgG paired cord blood; however, no IgM seropositivity was observed. All neonates had good Apgar scores and did not require NICU admission. Conclusion. In this study, 35.8% pregnant women were found to be dengue seropositive. However, transplacental transfer of IgG antibodies had no detrimental effect on the neonatal outcomes. PMID:25587564

  11. Antibody Discovery via Multiplexed Single Cell Characterization

    PubMed Central

    Harriman, William D.; Collarini, Ellen J.; Sperinde, Gizette V.; Strandh, Magnus; Fatholahi, Marjan M.; Dutta, April; Lee, Yunji; Mettler, Shelley E.; Keyt, Bruce A.; Ellsworth, Stote L.; Kauvar, Lawrence M.

    2009-01-01

    The secreted immunoglobulin footprint of single hybridoma cells, containing ~10 fg of antibody purified in situ, has been probed for 9 properties concurrently by use of detection labels comprising 280 nm combinatorially colored fluorescent latex beads functionalized with proteins. Specificity of each individual hybridoma cell’s product has thereby been assessed in a primary screen. Varying the density of antigen on beads to modulate the avidity of the interaction between bead and secreted antibody footprint allowed rank ordering by affinity in the same primary screen. As more criteria were added to the selection process, the frequency of positive cells went down; in some cases, the favorable cell was present at <1/50,000. Recovery of the cell of interest was accomplished by plating the cells in a viscous medium on top of a membrane. After collecting the antibody footprint on a capture surface beneath the membrane, the immobilized cells were transferred to an incubator while the footprints were analyzed to locate the hybridoma cells of interest. The desired cells were then cloned by picking them from the corresponding locations on the membrane. PMID:19087879

  12. Neutralizing monoclonal antibodies against listeriolysin: mapping of epitopes involved in pore formation.

    PubMed Central

    Darji, A; Niebuhr, K; Hense, M; Wehland, J; Chakraborty, T; Weiss, S

    1996-01-01

    Six different mouse monoclonal antibodies (MAbs) and a specific rabbit polygonal antibody were raised against listeriolysin. Four of the MAbs also recognized seeligeriolysin, and five cross-reacted with ivanolysin. The hemolytic activity could be neutralized by the polygonal antibody as well as by five of the MAbs. None of the neutralizing antibodies interfered with the binding of listeriolysin to the cellular membrane. The epitopes recognized by the MAbs were localized by using overlapping synthetic peptides between positions 59 and 279, a region hitherto not implicated in mediating hemolytic activity. PMID:8675351

  13. Radiolabeled antibodies in gynecologic tumors

    SciTech Connect

    Hardy, J.G.; Perkins, A.C.; Symonds, E.M.; Wastie, M.L.; Pimm, M.V.

    1984-01-01

    A monoclonal antibody has been raised against an osteogenic sarcoma cell line and radiolabeled with iodine-131. The antibody was administered to 12 patients with suspected ovarian tumors, two with recurrent carcinoma of the cervix and one with carcinoma of the body of the uterus. Each patient received an intravenous dose of 70 MBq I-131-labeled antibody and was imaged either 24 or 48 hours later. Image enhancement was achieved by subtraction of background activity using Tc-99m-labeled red blood cells and pertechnetate. In eleven patients with ovarian malignancies antibody uptake was detected at the suspected tumor sites, and agreed with the operative findings in the eight patients who subsequently underwent surgery. The patient in whom the antibody failed to localize was found to have a benign lesion. Uptake of antibody was seen at the tumor sites in the patients with carcinoma of the cervix and body of the uterus. The localization of tumor sites using I-131-labeled antibodies is difficult due to background activity, particularly from radioiodine in the bladder. In only five cases could the abnormal antibody concentration be identified on the iodine images alone. This problem was overcome by the use of background subtraction techniques. Immunoscintigraphy is proving useful for the assessment of tumor recurrence and as an aid to radiotherapy treatment planning.

  14. Renaissance of cancer therapeutic antibodies

    Microsoft Academic Search

    Martin J. Glennie

    2003-01-01

    In the past five years therapeutic monoclonal antibodies have established themselves as perhaps the most important and rapidly expanding class of therapeutic drugs. More than 25% of pharmacological agents that are currently under development are based on antibodies and the total income generated from them in 2002 exceeded $3 billion, and is predicted to rise to $10–20 billion by 2010.

  15. Prevalence of Herpes Simplex Virus Antibodies in Dental Students.

    ERIC Educational Resources Information Center

    Rodu, Brad; And Others

    1992-01-01

    A study of 125 sophomore preclinical dental students found that these young professionals, because of having a low prevalence of herpes simplex virus (HSV) antibodies, are at risk for acquiring a primary HSV infection when treating HSV positive patients and should take precautions to avoid virus transmission. (MSE)

  16. Delivery of antibodies to the cytosol: debunking the myths.

    PubMed

    Marschall, Andrea L J; Zhang, Congcong; Frenzel, André; Schirrmann, Thomas; Hust, Michael; Perez, Franck; Dübel, Stefan

    2014-01-01

    The use of antibodies to target their antigens in living cells is a powerful analytical tool for cell biology research. Not only can molecules be localized and visualized in living cells, but interference with cellular processes by antibodies may allow functional analysis down to the level of individual post-translational modifications and splice variants, which is not possible with genetic or RNA-based methods. To utilize the vast resource of available antibodies, an efficient system to deliver them into the cytosol from the outside is needed. Numerous strategies have been proposed, but the most robust and widely applicable procedure still remains to be identified, since a quantitative ranking of the efficiencies has not yet been done. To achieve this, we developed a novel efficiency evaluation method for antibody delivery based on a fusion protein consisting of a human IgG 1 Fc and the recombination enzyme Cre (Fc-Cre). Applied to suitable GFP reporter cells, it allows the important distinction between proteins trapped in endosomes and those delivered to the cytosol. Further, it ensures viability of positive cells and is unsusceptible to fixation artifacts and misinterpretation of cellular localization in microscopy and flow cytometry. Very low cytoplasmic delivery efficiencies were found for various profection reagents and membrane penetrating peptides, leaving electroporation as the only practically useful delivery method for antibodies. This was further verified by the successful application of this method to bind antibodies to cytosolic components in living cells. PMID:24848507

  17. Antineutrophil Cytoplasmic Antibodies, Autoimmune Neutropenia, and Vasculitis

    PubMed Central

    Grayson, Peter C.; Sloan, J. Mark; Niles, John L.; Monach, Paul A.; Merkel, Peter A.

    2011-01-01

    Objectives Reports of an association between antineutrophil cytoplasmic antibodies (ANCA) and autoimmune neutropenia have rarely included cases of proven vasculitis. A case of ANCA-associated vasculitis (AAV) with recurrent neutropenia is described and relevant literature on the association between ANCA, neutropenia, and vasculitis is reviewed. Methods Longitudinal clinical assessments and laboratory findings are described in a patient with AAV and recurrent episodes of profound neutropenia from December 2008 – October 2010. A PubMed database search of the medical literature was performed for papers published from 1960 through October 2010 to identify all reported cases of ANCA and neutropenia. Results A 49 year-old man developed recurrent neutropenia, periodic fevers, arthritis, biopsy-proven cutaneous vasculitis, sensorineural hearing loss, epididymitis, and positive tests for ANCA with specificity for antibodies to both proteinase 3 and myeloperoxidase. Antineutrophil membrane antibodies were detected during an acute neutropenic phase and were not detectable in a post-recovery sample, whereas ANCA titers did not seem to correlate with neutropenia. An association between ANCA and neutropenia has been reported in 74 cases from 24 studies in the context of drug/toxin exposure, underlying autoimmune disease, or chronic neutropenia without underlying autoimmune disease. In these cases, the presence of atypical ANCA patterns and other antibodies were common; however, vasculitis was uncommon and when it occurred was usually limited to the skin and in cases of underlying toxin exposure. Conclusions ANCA is associated with autoimmune neutropenia, but systemic vasculitis rarely occurs in association with ANCA and neutropenia. The interaction between neutrophils and ANCA may provide insight into understanding both autoimmune neutropenia and AAV. PMID:21507463

  18. Endogenous Antibodies for Tumor Detection

    PubMed Central

    Rich, Barrie S.; Honeyman, Joshua N.; Darcy, David G.; Smith, Peter T.; Williams, Andrew R.; Lim, Irene Isabel P.; Johnson, Linda K.; Gönen, Mithat; Simon, Joel S.; LaQuaglia, Michael P.; Simon, Sanford M.

    2014-01-01

    The study of cancer immunology has provided diagnostic and therapeutic instruments through serum autoantibody biomarkers and exogenous monoclonal antibodies. While some endogenous antibodies are found within or surrounding transformed tissue, the extent to which this exists has not been entirely characterized. We find that in transgenic and xenograft mouse models of cancer, endogenous gamma immunoglobulin (IgG) is present at higher concentration in malignantly transformed organs compared to non-transformed organs in the same mouse or organs of cognate wild-type mice. The enrichment of endogenous antibodies within the malignant tissue provides a potential means of identifying and tracking malignant cells in vivo as they mutate and diversify. Exploiting these antibodies for diagnostic and therapeutic purposes is possible through the use of agents that bind endogenous antibodies. PMID:24875800

  19. Abnormal measles-mumps-rubella antibodies and CNS autoimmunity in children with autism.

    PubMed

    Singh, Vijendra K; Lin, Sheren X; Newell, Elizabeth; Nelson, Courtney

    2002-01-01

    Autoimmunity to the central nervous system (CNS), especially to myelin basic protein (MBP), may play a causal role in autism, a neurodevelopmental disorder. Because many autistic children harbor elevated levels of measles antibodies, we conducted a serological study of measles-mumps-rubella (MMR) and MBP autoantibodies. Using serum samples of 125 autistic children and 92 control children, antibodies were assayed by ELISA or immunoblotting methods. ELISA analysis showed a significant increase in the level of MMR antibodies in autistic children. Immunoblotting analysis revealed the presence of an unusual MMR antibody in 75 of 125 (60%) autistic sera but not in control sera. This antibody specifically detected a protein of 73-75 kD of MMR. This protein band, as analyzed with monoclonal antibodies, was immunopositive for measles hemagglutinin (HA) protein but not for measles nucleoprotein and rubella or mumps viral proteins. Thus the MMR antibody in autistic sera detected measles HA protein, which is unique to the measles subunit of the vaccine. Furthermore, over 90% of MMR antibody-positive autistic sera were also positive for MBP autoantibodies, suggesting a strong association between MMR and CNS autoimmunity in autism. Stemming from this evidence, we suggest that an inappropriate antibody response to MMR, specifically the measles component thereof, might be related to pathogenesis of autism. PMID:12145534

  20. [The clinical informativeness of detection of antibodies to citrullinated proteins under rheumatoid arthritis].

    PubMed

    Cherkasova, M V; Novikov, A A; Alexndrova, E N; Karateev, D E; Popkova, T V; Luchikhina, E L; Avdeeva, A S; Nasonov, E L

    2015-02-01

    The main diagnostic laboratory markers of rheumatoid arthritis are IgM rheumatoid factor and antibodies to citrullinated proteins. The IgM rheumatoid factor is a sensitive but insufficiently specific marker of rheumatoid arthritis. The antibodies to citrullinated proteins have a higher specificity for diagnostic of rheumatoid arthritis. The antibodies to cyclic citrullinated peptide and modified citrullinated vimentin are the main representatives of family of antibodies to citrullinated proteins applying in clinical diagnostic practice. The study was carried out to deternine the role of antibodies to citrullinated proteins and modified citrullinated vimentin in diagnostic, evaluation of activity and severity of destructive alterations under rheumatoid arthritis. The samplings of 993 patients with reliable diagnosis of rheumatoid arthritis. 179 patients with other rheumatoid diseases and 30 healthy donors were examined. The measurement of serum concentration of IgM rheumatoid factor and C-reactive protein was implemented by immune nephelometric analysis and antibodies to citrullinated proteins were analyzed by enzymoimmunoassay The erythrocyte sedimentation rate was established using the Westergreen technique. It was established that antibodies to modified citrullinated vimentin had the highest diagnostic specificity (83%), antibodies to cyclic citrullinated peptide had the highest diagnostic specificity (87%). The diagnostic specificity of joint detection of IgM rheumatoid factor, antibodies to citrullinated proteins and antibodies to modified citrullinated vimentin made up to 87%. In patients negative to rheumatoid factor the rate ofdetection of antibodies to citrullinated proteins made up to 34% and antibodies to modified citrullinated vimentin made up to 48%. The diagnostic effectiveness of detection of antibodies to citrullinitted proteins (ratio of likelihood of positive and negative results of test was correspondingly 5.5 and 0.3; area under ROC curve 0.8) and antibodies to modified citrullinated vimentin (ratio of likelihood of positive and negative results of test was correspondingly 4.4 and 0.2; area under ROC curve 0.9) surpassed the same in analysis of IgM rheumatoid factor (ratio of likelihood of positive results--3.2, ratio of likelihood of negative results--0.4, area under ROC curve--0.8). The weak positive correlation relationship was established between concentration of antibodies to cyclic citrillinatedpeptide/antibodies to modified citrullinated vimentin in blood serum and indicators of clinical laboratory activity of rheumatoid arthritis (ESR, CRP DAS 28, (r-0.2. p < 0.05). The high positive levels of antibodies to modified citrullinated vimentin associated with expressed destructive affection of joints (p < 0.02). The antibodies to cyclic citrullinated peptide are the most highly specific and clinically informative laboratory diagnostic marker of rheumatoid arthritis. The detection of antibodies to modified citrullinated vimentin is an important additional serological test to diagnose rheumatoid arthritis in IgM rheumatoid factor-negative and/or antibodies to cyclic citrullinated peptide-negative patients and to forecast severe destructive affection of joints under the given disease. The joint study of IgM rheumatoid factor, antibodies to cyclic citrullinated peptide and antibodies to modified citrullinated vimentin under rheumatoid arthritis has higher diagnostic sensitivity as compared with isolated antibodies to citrullinated proteins. PMID:26027260

  1. Neonatal alloimmune thrombocytopenia due to anti-human leukocyte antigen antibody: a case report.

    PubMed

    Sasaki, M; Yagihashi, A; Kobayashi, D; Watanabe, N; Fujikawa, T; Chiba, S; Sato, S; Morishita, K; Sekimoto, T; Ikeda, H

    2001-12-01

    Anti-HLA antibodies reportedly exist in 31% of pregnant women. However, few ocurrences of neonatal alloimmune thrombocytopenia (NAIT) caused by anti-HLA antibody have been reported. In this study, maternal anti-HLA B60 and B61 antibodies were identified in patient serum at birth, but no anti-platelet antibodies were present. No maternal anti-HLA A2, A24, B51, or B52 antibodies were detected in patient serum. Platelet transfusion from the third donor was effective because these platelets expressed HLA A24 and B52 but not B60 or B61. Cross-matching tests between patient leukocytes or platelets and maternal serum were strongly positive, indicating that maternal anti-HLA antibodies were responsible for NAIT. This report is the first to demonstrate NAIT probably caused by maternal anti-HLA A24 and B52. PMID:11764101

  2. A Unique Human Mycoplasma Protein that Generically Blocks Antigen-Antibody Union

    PubMed Central

    Nieusma, Travis; Jones, Teresa; Boreo, Isabel; MacLeod, Amanda S.; Mark, Adam; Niessen, Sherry; Kim, Helen J.; Kong, Leopold; Assad-Garcia, Nacyra; Kwon, Keehwan; Chesi, Marta; Smider, Vaughn V.; Salomon, Daniel R.; Jelinek, Diane F.; Kyle, Robert A.; Pyles, Richard B.; Glass, John I.; Ward, Andrew B.; Wilson, Ian A.; Lerner, Richard A.

    2014-01-01

    We report the discovery and crystal structure of a human mycoplasma protein, Protein M, which binds with high affinity to antibodies, predominantly through attachment to the variable region of the ? and ? light chains. Protein M broadly blocks antibody-antigen union and its mechanism of inhibition is of considerable interest because, as a diversity system, the binding mode of each antibody is different. Protein M thus appears to function by a mechanism that is independent of the sequences of members of the extensive antibody repertoire. By anchoring to conserved regions of the antibody light chains, Protein M is in a position to extend its large C-terminal domain over the antibody combining site and block entrance to macromolecular antigens. PMID:24503852

  3. Antibodies targeting mutated citrullinated vimentin in patients with psoriatic arthritis.

    PubMed

    Tesija-Kuna, Andrea; Grazio, Simeon; Miler, Marijana; Vukasovic, Ines; Peric, Porin; Vrkic, Nada

    2010-05-01

    Antibodies against mutated citrullinated vimentin (anti-MCV) are of a comparable diagnostic value in rheumatoid arthritis (RA) as antibodies targeting citrullinated peptides (anti-CCP). Anti-CCP are present in up to 15% of psoriatic arthritis (PsA) patients, while the prevalence of anti-MCV in PsA patients has been poorly investigated. The aim of the present study was to assess the prevalence and relevance of anti-MCV antibodies in PsA patients. The study included 56 PsA patients. Clinical features, disease activity, and functional ability were noted by an experienced rheumatologist. Serum samples of all patients were analyzed for anti-MCV and anti-CCP antibodies using enzyme-linked immunosorbent assay. Data on 92 patients with RA, 44 patients with other inflammatory rheumatic diseases, and 107 healthy controls from a previous study were used to compare the prevalence of anti-MCV antibodies in PsA patients. Anti-MCV antibodies were positive in only two out of 56 (3.6%) PsA patients, which was significantly lower compared to RA patients (63%). The anti-MCV level was moderately positive and borderline in one patient each. Both patients had asymmetric polyarthritis, dactylitis, moderate to high disease activity, and were anti-CCP and rheumatoid factor (RF) negative. There was no significant difference in anti-MCV levels according to clinical subtypes of PsA and no correlation of anti-MCV levels with anti-CCP, RF, disease activity variables, and functional ability indices. According to study results, anti-MCV antibodies can be detected in a very small proportion of PsA patients with polyarthritic disease and are primarily related to the polyarthritic pattern rather than the specific diagnosis of RA. PMID:20069329

  4. RosettaAntibody: antibody variable region homology modeling server.

    PubMed

    Sircar, Aroop; Kim, Eric T; Gray, Jeffrey J

    2009-07-01

    The RosettaAntibody server (http://antibody.graylab.jhu.edu) predicts the structure of an antibody variable region given the amino-acid sequences of the respective light and heavy chains. In an initial stage, the server identifies and displays the most sequence homologous template structures for the light and heavy framework regions and each of the complementarity determining region (CDR) loops. Subsequently, the most homologous templates are assembled into a side-chain optimized crude model, and the server returns a picture and coordinate file. For users requesting a high-resolution model, the server executes the full RosettaAntibody protocol which additionally models the hyper-variable CDR H3 loop. The high-resolution protocol also relieves steric clashes by optimizing the CDR backbone torsion angles and by simultaneously perturbing the relative orientation of the light and heavy chains. RosettaAntibody generates 2000 independent structures, and the server returns pictures, coordinate files, and detailed scoring information for the 10 top-scoring models. The 10 models enable users to use rational judgment in choosing the best model or to use the set as an ensemble for further studies such as docking. The high-resolution models generated by RosettaAntibody have been used for the successful prediction of antibody-antigen complex structures. PMID:19458157

  5. Micromechanical antibody sensor

    DOEpatents

    Thundat, Thomas G. (Knoxville, TN); Jacobson, K. Bruce (Oak Ridge, TN); Doktycz, Mitchel J. (Knoxville, TN); Kennel, Stephen J. (Oak Ridge, TN); Warmack, Robert J. (Knoxville, TN)

    2001-01-01

    A sensor apparatus is provided using a microcantilevered spring element having a coating of a detector molecule such as an antibody or antigen. A sample containing a target molecule or substrate is provided to the coating. The spring element bends in response to the stress induced by the binding which occurs between the detector and target molecules. Deflections of the cantilever are detected by a variety of detection techniques. The microcantilever may be approximately 1 to 200 .mu.m long, approximately 1 to 50 .mu.m wide, and approximately 0.3 to 3.0 .mu.m thick. A sensitivity for detection of deflections is in the range of 0.01 nanometers.

  6. Prevalence of antibodies to canine parvovirus and distemper virus in wolves in the Canadian Rocky Mountains.

    PubMed

    Nelson, Brynn; Hebblewhite, Mark; Ezenwa, Vanessa; Shury, Todd; Merrill, Evelyn H; Paquet, Paul C; Schmiegelow, Fiona; Seip, Dale; Skinner, Geoff; Webb, Nathan

    2012-01-01

    Wild carnivores are often exposed to diseases via contact with peridomestic host species that travel through the wildland-urban interfaces. To determine the antibody prevalences and relationships to human activity for two common canid pathogens, we sampled 99 wolves (Canis lupus) from 2000 to 2008 for antibodies to canine parvovirus (CPV) and canine distemper virus (CDV) in Banff and Jasper National Parks and surrounding areas of the Canadian Rockies. This population was the source for wolves reintroduced into the Northern Rockies of the US. Of 99 wolves sampled, 94 had detectable antibody to CPV (95%), 24 were antibody-positive for CDV (24%), and 24 had antibodies to both pathogens (24%). We tested whether antibody prevalences for CPV and CDV were higher closer to human activity (roads, town sites, First Nation reserves) and as a function of sex and age class. Wolves ?2 yr old were more likely to be have antibodies to CPV. For CDV, male wolves, wolves ?2 yr, and those closer to First Nation reserves were more likely to have antibodies. Overall, however, we found minimal support for human influence on antibody prevalence for CDV and CPV. The similarity between our antibody prevalence results and results from recent studies in Yellowstone National Park suggests that at least in the case of CDV, and perhaps CPV, these could be important pathogens with potential effects on wolf populations. PMID:22247375

  7. Fully Human Antagonistic Antibodies against CCR4 Potently Inhibit Cell Signaling and Chemotaxis

    PubMed Central

    Géraudie, Solène; Scheffler, Ulrike; Griep, Remko A.; Reiersen, Herald; Duncan, Alexander R.; Kiprijanov, Sergej M.

    2014-01-01

    Background CC chemokine receptor 4 (CCR4) represents a potentially important target for cancer immunotherapy due to its expression on tumor infiltrating immune cells including regulatory T cells (Tregs) and on tumor cells in several cancer types and its role in metastasis. Methodology Using phage display, human antibody library, affinity maturation and a cell-based antibody selection strategy, the antibody variants against human CCR4 were generated. These antibodies effectively competed with ligand binding, were able to block ligand-induced signaling and cell migration, and demonstrated efficient killing of CCR4-positive tumor cells via ADCC and phagocytosis. In a mouse model of human T-cell lymphoma, significant survival benefit was demonstrated for animals treated with the newly selected anti-CCR4 antibodies. Significance For the first time, successful generation of anti- G-protein coupled chemokine receptor (GPCR) antibodies using human non-immune library and phage display on GPCR-expressing cells was demonstrated. The generated anti-CCR4 antibodies possess a dual mode of action (inhibition of ligand-induced signaling and antibody-directed tumor cell killing). The data demonstrate that the anti-tumor activity in vivo is mediated, at least in part, through Fc-receptor dependent effector mechanisms, such as ADCC and phagocytosis. Anti-CC chemokine receptor 4 antibodies inhibiting receptor signaling have potential as immunomodulatory antibodies for cancer. PMID:25080123

  8. Catch Me If You Can - The Race Between HIV and Neutralizing Antibodies.

    PubMed

    Geiß, Yvonne; Dietrich, Ursula

    2015-01-01

    Broadly neutralizing antibodies represent the major protective mechanism of vaccines targeting pathogenic microbes in humans and animals. For HIV, broadly neutralizing antibodies have also been shown to be protective in experimental animal models. However, despite the identification of a respectable number of broadly neutralizing antibodies from chronically infected HIV-positive persons in recent years, attempts to induce such antibodies by vaccines have generally failed over the last decades. Though unsuccessful in view of achieving a protective vaccine against HIV, many of these studies have contributed significantly to the understanding of the generation of broadly neutralizing antibodies against HIV-1 as well as to the vulnerable sites they target on the surface of the virus. Here we review the most important features of patient-derived broadly neutralizing antibodies, the long and complex B-cell maturation pathways required for their production, and the resulting consequences for vaccine development. We further address characteristics of the epitopes targeted by broadly neutralizing antibodies on the virus surface as well as mechanisms of viral escape. Taken together, the identification of vaccine candidates able to induce broadly neutralizing antibodies against HIV-1 is the major challenge in HIV vaccine development. Mutual coevolution of rationally designed HIV vaccine candidates, with affinity maturation pathways of antibodies they induce upon vaccination, may best mimic the natural situation of chronically HIV-infected patients who are able to generate broadly neutralizing antibodies. PMID:26035168

  9. EBV transformation and microfusion as the potential source of human monoclonal antisperm antibodies.

    PubMed

    Kurpisz, M; Simon, L L; Alexander, N J

    1987-10-01

    Peripheral blood lymphocytes were obtained from vasectomized men with high serum titers of antisperm antibodies. An Epstein-Barr virus (EBV) transformation was performed either with B cells or mononuclear leukocytes. The effect of feeder cells (irradiated umbilical cord blood lymphocytes), cyclosporin A, and in vitro stimulation of lymphocytes with sperm extract on EBV transformation was evaluated. Antibody-producing cells were screened for specificity against human sperm by an enzyme-linked immunosorption assay (ELISA) one to six weeks after transformation. Using B cells or leukocyte mononuclear cells, we found that the percentage of wells containing antibody reactive against human sperm was greatest two weeks after transformation (range 3% to 7.5% positive wells). To increase and maintain antibody synthesis by these transformed cells, microfusions were performed in those wells positive for antisperm antibody using the UC 729-6 lymphoblastoid cell partner. Then resultant hybridomas were expanded, subcloned, and preliminarily characterized. PMID:2829635

  10. Antibodies to Pf155, a major antigen of Plasmodium falciparum: seroepidemiological studies in Haiti*

    PubMed Central

    Deloron, P.; Duverseau, Y. T.; Zevallos-Ipenza, A.; Magloire, R.; Stanfill, P. S.; Nguyen-Dinh, Phuc

    1987-01-01

    The presence of malaria parasites and the serological antibody responses against whole Plasmodium falciparum and the Pf155 antigen were studied in the population of a small rural locality in Haiti in December 1985. Only 7 (1.5%) of the individuals were found to be infected with P. falciparum, the only species observed. Antibodies to P. falciparum were detected in an ELISA in 38.2% of the sera, the positivity rates being age-related. Anti-Pf155 antibodies were detected in 12.5% and 13.6% of individuals by two different techniques used. The anti-Pf155 positivity rates increased only after 25 years of age. No trends were detected for a clear-cut protective value of Pf155 antibodies against clinical malaria and further longitudinally conducted field surveys are needed to satisfactorily assess the potential protective effect of Pf155 antibodies. ImagesFig. 2 PMID:3311436

  11. Comparison of serological tests for detection of antibodies to infectious laryngotracheitis virus

    Microsoft Academic Search

    B. M. Adair; D. Todd; E. R. McKillop; K. Burns

    1985-01-01

    Four serological tests i.e. ELISA, serum neutralisation (SN), fluorescent antibody (FA), and agar gel immunodiffusion (AGID) were compared for sensitivity using several criteria, for detection and titration of infectious laryngotracheitis (ILT) virus antibodies in chicken sera. In the ELISA test, sera were tested in parallel on virus positive and negative control antigens with results expressed as positive?negative difference. Non?specific binding

  12. Adipose tissue development in the fetal pig examined using monoclonal antibodies

    Microsoft Academic Search

    J. T. Wright; G. J. Hausman

    ABSTRACT Two anti-adipocyte monoclonal,antibodies (MAbs; AD-I and AD-2) have been used to study the development,of dorsal S.C. adipose tissue in fetuses from 50 to 110 d of gestation. Immunofluorescent staining of cryostat sections with each antibody,revealed anhgen- positive cells in fetal S.C. mesenchyme,prior to lipid deposition. Lipid droplets as well as AD-I and AD-2 positive cells were detected within the

  13. Restricted diversity of antigen binding residues of antibodies revealed by computational alanine scanning of 227 antibody-antigen complexes.

    PubMed

    Robin, Gautier; Sato, Yoshiteru; Desplancq, Dominique; Rochel, Natacha; Weiss, Etienne; Martineau, Pierre

    2014-11-11

    Antibody molecules are able to recognize any antigen with high affinity and specificity. To get insight into the molecular diversity at the source of this functional diversity, we compiled and analyzed a non-redundant aligned collection of 227 structures of antibody-antigen complexes. Free energy of binding of all the residue side chains was quantified by computational alanine scanning, allowing the first large-scale quantitative description of antibody paratopes. This demonstrated that as few as 8 residues among 30 key positions are sufficient to explain 80% of the binding free energy in most complexes. At these positions, the residue distribution is not only different from that of other surface residues but also dependent on the role played by the side chain in the interaction, residues participating in the binding energy being mainly aromatic residues, and Gly or Ser otherwise. To question the generality of these binding characteristics, we isolated an antibody fragment by phage display using a biased synthetic repertoire with only two diversified complementarity-determining regions and solved its structure in complex with its antigen. Despite this restricted diversity, the structure demonstrated that all complementarity-determining regions were involved in the interaction with the antigen and that the rules derived from the natural antibody repertoire apply to this synthetic binder, thus demonstrating the robustness and universality of our results. PMID:25174334

  14. Satellite positioning

    NASA Technical Reports Server (NTRS)

    Colombo, Oscar L.; Watkins, Michael M.

    1991-01-01

    Developments in satellite positioning techniques and their applications are reviewed on the basis of the theoretical and practical work published by U.S. researchers in 1987-1990. Current techniques are classified into two main categories: satellite laser tracking and radio tracking. Particular attention is given to the Geoscience Laser Ranging System, the Lunar Laser Ranging concept; GPS ephemerides determination, fiducial networks, and reference frame; static GPS positioning; and kinematic GPS positioning.

  15. Encephalitis and GABAB receptor antibodies

    PubMed Central

    Höftberger, Romana; Titulaer, Maarten J.; Sabater, Lidia; Dome, Balazs; Rózsás, Anita; Hegedus, Balazs; Hoda, Mir Alireza; Laszlo, Viktoria; Ankersmit, Hendrik Jan; Harms, Lutz; Boyero, Sabas; de Felipe, Alicia; Saiz, Albert; Dalmau, Josep

    2013-01-01

    Objective: To report the clinical features of 20 newly diagnosed patients with GABAB receptor (GABABR) antibodies and determine the frequency of associated tumors and concurrent neuronal autoantibodies. Methods: Clinical data were retrospectively obtained and evaluated. Serum and CSF samples were examined for additional antibodies using methods previously reported. Results: Seventeen patients presented with seizures, memory loss, and confusion, compatible with limbic encephalitis (LE), one patient presented with ataxia, one patient presented with status epilepticus, and one patient presented with opsoclonus-myoclonus syndrome (OMS). Nineteen (95%) patients eventually developed LE during the course of the disease. Small-cell lung cancer (SCLC) was identified in 10 (50%) patients, all with LE. Treatment and outcome was available from 19 patients: 15 showed complete (n = 7) or partial (n = 8) neurologic improvement after steroids, IV immunoglobulins, or plasma exchange and oncologic treatment when indicated; 1 patient died of tumor progression shortly after the first cycle of immunotherapy, and 3 were not treated. Five patients with SCLC had additional onconeuronal antibodies (Ri, amphiphysin, or SOX1), and 2 without tumor had GAD65 and NMDAR antibodies, respectively. GABABR antibodies were not detected in serum of 116 patients with SCLC without neurologic symptoms. Conclusion: Our study confirms GABABR as an autoantigen of paraneoplastic and nonparaneoplastic LE and expands the phenotype of GABABR antibodies to ataxia, OMS, and status epilepticus. The long-term prognosis is dictated by the presence of a tumor. Recognition of syndromes associated with GABABR antibodies is important because they usually respond to treatment. PMID:24068784

  16. CATNAP: a tool to compile, analyze and tally neutralizing antibody panels.

    PubMed

    Yoon, Hyejin; Macke, Jennifer; West, Anthony P; Foley, Brian; Bjorkman, Pamela J; Korber, Bette; Yusim, Karina

    2015-07-01

    CATNAP (Compile, Analyze and Tally NAb Panels) is a new web server at Los Alamos HIV Database, created to respond to the newest advances in HIV neutralizing antibody research. It is a comprehensive platform focusing on neutralizing antibody potencies in conjunction with viral sequences. CATNAP integrates neutralization and sequence data from published studies, and allows users to analyze that data for each HIV Envelope protein sequence position and each antibody. The tool has multiple data retrieval and analysis options. As input, the user can pick specific antibodies and viruses, choose a panel from a published study, or supply their own data. The output superimposes neutralization panel data, virus epidemiological data, and viral protein sequence alignments on one page, and provides further information and analyses. The user can highlight alignment positions, or select antibody contact residues and view position-specific information from the HIV databases. The tool calculates tallies of amino acids and N-linked glycosylation motifs, counts of antibody-sensitive and -resistant viruses in conjunction with each amino acid or N-glycosylation motif, and performs Fisher's exact test to detect potential positive or negative amino acid associations for the selected antibody. Website name: CATNAP (Compile, Analyze and Tally NAb Panels). Website address: http://hiv.lanl.gov/catnap. PMID:26044712

  17. CATNAP: a tool to compile, analyze and tally neutralizing antibody panels

    PubMed Central

    Yoon, Hyejin; Macke, Jennifer; West, Anthony P.; Foley, Brian; Bjorkman, Pamela J.; Korber, Bette; Yusim, Karina

    2015-01-01

    CATNAP (Compile, Analyze and Tally NAb Panels) is a new web server at Los Alamos HIV Database, created to respond to the newest advances in HIV neutralizing antibody research. It is a comprehensive platform focusing on neutralizing antibody potencies in conjunction with viral sequences. CATNAP integrates neutralization and sequence data from published studies, and allows users to analyze that data for each HIV Envelope protein sequence position and each antibody. The tool has multiple data retrieval and analysis options. As input, the user can pick specific antibodies and viruses, choose a panel from a published study, or supply their own data. The output superimposes neutralization panel data, virus epidemiological data, and viral protein sequence alignments on one page, and provides further information and analyses. The user can highlight alignment positions, or select antibody contact residues and view position-specific information from the HIV databases. The tool calculates tallies of amino acids and N-linked glycosylation motifs, counts of antibody-sensitive and -resistant viruses in conjunction with each amino acid or N-glycosylation motif, and performs Fisher's exact test to detect potential positive or negative amino acid associations for the selected antibody. Website name: CATNAP (Compile, Analyze and Tally NAb Panels). Website address: http://hiv.lanl.gov/catnap. PMID:26044712

  18. Toxoplasma antibodies among bobcats and other carnivores of norther California.

    PubMed

    Riemann, H P; Howarth, J A; Ruppanner, R; Franti, C E; BEHYMER, D E

    1975-04-01

    The prevalence of antibodies to Toxoplasma gondii was investigated among five species of wild carnivores in Norther Ccalifornia. The highest prevalence was among bobcats (Lynx rufus), with 15 of 21 tested being serologically positive. Other results included serological evidence of toxoplasmosis in two of seven raccoons (Procyon lotor), one of three badgers (taxidea taxus) and two of three coyotes (Canis latrans). Two gray foxes (Urocyon cinereoargenteus) were serologically negative. Oone badger with an indirect hemagglutination antibody titer of 1:8192 was found to harbor T. gondii in its brain tissues. PMID:1142562

  19. Toxoplasma gondii antibodies in exotic wild felids from Brazilian zoos.

    PubMed

    Silva, J C; Ogassawara, S; Marvulo, M F; Ferreira-Neto, J S; Dubey, J P

    2001-09-01

    Serum samples from 37 captive exotic felids in 12 zoos from six Brazilian states were assayed for antibodies to Toxoplasma gondii by the modified agglutination test using formalin-fixed whole tachyzoites. Titers greater than or equal to 1:20 were considered positive. Antibodies to T. gondii were found in 24 of 37 (64.9%) felids, including one European lynx (Lynx lynx), two jungle cats (Felis chaus), two servals (Leptailurus serval), two tigers (Panthera tigris), three leopards (Panthera pardus), and 14 of 27 lions (Panthera leo). This is the first serologic analysis for T. gondii infection in exotic wild felids from Brazilian zoos. PMID:12785684

  20. Modified antibody in fetal alloimmunization.

    PubMed

    Bussel, James B; McFarland, Janice G

    2013-07-18

    In this issue of Blood, Ghevaert et al propose to develop a therapeutic antibody for fetal and neonatal alloimmune thrombocytopenia (FNAIT) that would block the actual antibody in sensitized mothers from binding and therefore prevent, or at least ameliorate, fetal and neonatal thrombocytopenia in fetuses who would otherwise be affected.1 The goal of the group is to engineer an antibody reagent that would on the one hand not engage conventional activating Fc receptors and on the other hand interact normally with FcRn, allowing transplacental passage. PMID:23869072

  1. Reducing heterophilic antibody interference in immunoassays using single chain antibodies

    SciTech Connect

    Baird, Cheryl L.; Tan, Ruimin; Fischer, Christopher J.; Victry, Kristin D.; Zangar, Richard C.; Rodland, Karin D.

    2011-12-15

    Sandwich ELISA microarrays have the potential to simultaneously quantify the levels of multiple diagnostic targets in a biological sample. However, as seen with traditional ELISA diagnostics, heterophilic antibodies (HA) in patient sera have the potential to cause interference in these assays. We demonstrate here that reducing the diagnostic capture antibody to its minimal functional unit, the variable heavy and light domains artificially connected with a short polypeptide linker (scFv), is an effective strategy for reducing the HA assay interference.

  2. Mutations conferring resistance to neutralization with monoclonal antibodies in type 1 poliovirus can be located outside or inside the antibody-binding site.

    PubMed Central

    Blondel, B; Crainic, R; Fichot, O; Dufraisse, G; Candrea, A; Diamond, D; Girard, M; Horaud, F

    1986-01-01

    Antigenic variants resistant to eight neutralizing monoclonal antibodies were selected from wild (Mahoney) and attenuated (Sabin) type 1 infectious poliovirions. Cross-immunoprecipitation revealed interrelationships between epitopes which were not detected by cross-neutralization. Operational analysis of antigenic variants showed that seven of eight neutralization epitopes studied were interrelated. Only one neutralization epitope, named Kc, varied independently from all the others. This latter, recognized by C3 neutralizing monoclonal antibody, was present not only on infectious virions but also on heat-denatured (C-antigenic) particles and on isolated capsid protein VP1. Loss of the neutralization function of an epitope did not necessary result from the loss of its antibody-binding capacity. Such potential, but not functional, neutralization epitopes exist naturally on Mahoney and Sabin 1 viruses. Their antibody-binding property could be disrupted by isolating antigenic variants in the presence of the nonneutralizing monoclonal antibody and anti-mouse immunoglobulin antibodies. Single-point mutations responsible for the acquisition of resistance to neutralization in the antigenic variants were located by sequence analyses of their genomes. Mutants selected in the presence of C3 neutralizing monoclonal antibody always had the mutation located inside the antibody-binding site (residues 93 through 103 of VP1) at the amino acid position 100 of VP1. On the contrary, antigenic variants selected in the presence of neutralizing monoclonal antibodies reacting only with D-antigenic particles had mutations situated in VP3, outside the antibody-binding site (residues 93 through 103 of VP1). The complete conversion of the Mahoney to the Sabin 1 epitope map resulted from a threonine-to-lysine substitution at position 60 of VP3. PMID:2416957

  3. Human anti-luteinizing hormone-releasing hormone antibodies in patients treated with synthetic luteinizing hormone-releasing hormone

    SciTech Connect

    Meakin, J.L.; Keogh, E.J.; Martin, C.E.

    1985-05-01

    One hundred sixty-three patients who were given synthetic LH-RH therapeutically underwent monitoring of serum IgG anti-LH-RH antibodies. Five of the patients showed specific binding to antibodies. Development of anti-LH-RH antibodies was not limited to those patients with a congenital deficiency of LH-RH. Urticarial responses occurred in four patients, only one of whom had IgG antibodies. Patients who had IgG antibodies or an urticarial response underwent monitoring of their serum IgE anti-LH-RH antibodies, but none had a positive binding response. The refractory state which has been reported in patients in whom similar antibodies to LH-RH develop was not invariably observed among these patients.

  4. Structural Biology of Moonlighting --Lessons from Antibodies

    E-print Network

    Martin, Andrew C.R.

    Structural Biology of Moonlighting -- Lessons from Antibodies Andrew C.R. Martin Institute that they are both part of what an antibody needs to do. Nonetheless, antibodies provide interesting lessons fragment) forms the stem of the Y-shape and is responsible for the effector functions of the antibody

  5. Structural Biology of Moonlighting --Lessons from Antibodies

    E-print Network

    Martin, Andrew C.R.

    Structural Biology of Moonlighting -- Lessons from Antibodies Andrew C.R. Martin Institute that they are both part of what an antibody needs to do. Nonetheless, antibodies provide interesting lessons the stem of the Y-shape and is responsible for the effector functions of the antibody such as binding to F

  6. Association of antithyroid peroxidase antibody with fibromyalgia in rheumatoid arthritis.

    PubMed

    Ahmad, Jowairiyya; Blumen, Helena; Tagoe, Clement E

    2015-08-01

    To investigate how autoimmune thyroiditis (ATD) affects the clinical presentation of established rheumatoid arthritis (RA) with particular reference to fibromyalgia and chronic widespread pain (CWP). A cohort of 204 patients with RA for whom the presence or absence of autoimmune thyroid antibodies was documented was examined for the relationships between thyroid autoantibodies and fibromyalgia or CWP. We identified 29 % who tested positive for antithyroid peroxidase antibodies (TPOAb). The anti-thyroglobulin antibody (TgAb) was found in 24 %. Among the thyroid autoantibody-positive patients, 40 % had a diagnosis of fibromyalgia or CWP versus 17 % for antibody negative patients. Logistic regression analyses (adjusted by age, sex, diabetes and BMI) indicated that TPOAb-positive patients were more likely to have fibromyalgia or CWP, with an odds ratio (OR) of 4.641, 95 % confidence interval (CI) (2.110-10.207) P < .001. Adjusting for spinal degenerative disc disease did not change the association with fibromyalgia, OR 4.458, 95 % CI (1.950-10.191), P < .001. The OR between TgAb and fibromyalgia was not significant (P > .05). Additional logistic regression analyses (adjusted by age, sex and BMI) indicated a significant relationship between TPOAb and fibromyalgia or CWP in patients without diabetes and those without hypothyroidism (OR of 4.873, 95 % CI (1.877-12.653), P = .001 and OR of 4.615 95 % CI (1.810-11.770), P = .001, respectively). There may be a positive association between the ATD antibody TPOAb, and fibromyalgia syndrome and CWP in patients with established RA. PMID:25976307

  7. Surface chemistries for antibody microarrays

    SciTech Connect

    Seurynck-Servoss, Shannon L.; Baird, Cheryl L.; Rodland, Karin D.; Zangar, Richard C.

    2007-05-01

    Enzyme-linked immunosorbent assay (ELISA) microarrays promise to be a powerful tool for the detection of disease biomarkers. The original technology for printing ELISA microarray chips and capturing antibodies on slides was derived from the DNA microarray field. However, due to the need to maintain antibody structure and function when immobilized, surface chemistries used for DNA microarrays are not always appropriate for ELISA microarrays. In order to identify better surface chemistries for antibody capture, a number of commercial companies and academic research groups have developed new slide types that could improve antibody function in microarray applications. In this review we compare and contrast the commercially available slide chemistries, as well as highlight some promising recent advances in the field.

  8. Antiphospholipid-antibody-associated panniculitis.

    PubMed

    Hunt, Raegan D; Robinson, Maria; Patel, Rishi; Franks, Andrew G

    2012-12-01

    A 60-year-old man presented with intermittent, tender, erythematous nodules on the legs that were associated with mild arthralgias. He was otherwise asymptomatic but reported a history of lupus anticoagulant antibodies that were discovered incidentally on laboratory screening at the approximate time that his lesions first occurred. A biopsy specimen showed a septal and lobular panniculitis with neutrophils, histiocytes, numerous eosinophils, foci of fibrosis, and fat necrosis but no vascular pathology. An elevated activated partial thromboplastin time (PTT), appreciably elevated levels of anti-beta-2 glycoprotein I antibody (IgM and IgG), and moderately elevated levels of anticardiolipin antibody (IgM and IgG) were present. The onset and recurrence of his skin condition coincided with increased antiphospholipid antibody levels and treatment with 81 mg aspirin daily was associated with improvement. PMID:23286808

  9. Biophysical Signatures of Monoclonal Antibodies

    Microsoft Academic Search

    N. Harn; T. Spitznagel; M. Perkins; C. Allan; S. Shire; C. R. Middaugh

    \\u000a Monoclonal antibodies are the most common protein that is being developed by many companies as therapies against a wide range\\u000a of diseases (Andreakos et al. 2002; Campbell and Marcus 2003; Trikha et al. 2002; Untch et al. 2003). With increasing interest\\u000a in the use of monoclonal antibodies therapeutics, it is apparent that the combination of specificity and safety offered by

  10. Radioimmunoguided surgery using monoclonal antibody

    Microsoft Academic Search

    E. W. Jr. Martin; C. M. Mojzisik; G. H. Jr. Hinkle; J. Sampsel; M. A. Siddiqi; S. E. Tuttle; B. Sickle-Santanello; D. Colcher; M. O. Thurston; J. G. Bell

    1988-01-01

    The potential proficiency of radioimmunoguided surgery in the intraoperative detection of tumors was assessed using labeled monoclonal antibody B72.3 in 66 patients with tissue-proved tumor. Monoclonal antibody B72.3 was injected 5 to 42 days preoperatively, and the hand-held gamma-detecting probe was used intraoperatively to detect the presence of tumor. Intraoperative probe counts of less than 20 every 2 seconds, or

  11. Therapeutic Anti-VEGF Antibodies

    Microsoft Academic Search

    S. Lien; H. B. Lowman

    Vascular endothelial growth factor (VEGF-A) is a key cytokine in the development of normal blood vessels as well as the development\\u000a of vessels in tumors and other tissues undergoing abnormal angiogenesis. Here, we review the molecular engineering of two\\u000a humanized antibodies derived from a common mouse anti-VEGF antibody — bevacizumab, a full-length IgG1 approved for the treatment\\u000a of specified cancer

  12. Nuclear Positioning

    PubMed Central

    Gundersen, Gregg G.; Worman, Howard J.

    2013-01-01

    SUMMARY The nucleus is the largest organelle and is commonly depicted in the center of the cell. Yet during cell division, migration and differentiation, it frequently moves to an asymmetric position aligned with cell function. We consider the toolbox of proteins that move and anchor the nucleus within the cell and how forces generated by the cytoskeleton are coupled to the nucleus to move it. The significance of proper nuclear positioning is underscored by numerous diseases resulting from genetic alterations in the toolbox proteins. Finally, we discuss how nuclear position may influence cellular organization and signaling pathways. PMID:23498944

  13. Humanised IgG1 antibody variants targeting membrane-bound carcinoembryonic antigen by antibody-dependent cellular cytotoxicity and phagocytosis

    PubMed Central

    Ashraf, S Q; Umana, P; Mössner, E; Ntouroupi, T; Brünker, P; Schmidt, C; Wilding, J L; Mortensen, N J; Bodmer, W F

    2009-01-01

    Background: The effect of glycoengineering a membrane specific anti-carcinoembryonic antigen (CEA) (this paper uses the original term CEA for the formally designated CEACAM5) antibody (PR1A3) on its ability to enhance killing of colorectal cancer (CRC) cell lines by human immune effector cells was assessed. In vivo efficacy of the antibody was also tested. Methods: The antibody was modified using EBNA cells cotransfected with ?-1,4-N-acetylglucosaminyltransferase III and the humanised hPR1A3 antibody genes. Results: The resulting alteration of the Fc segment glycosylation pattern enhances the antibody's binding affinity to the Fc?RIIIa receptor on human immune effector cells but does not alter the antibody's binding capacity. Antibody-dependent cellular cytotoxicity (ADCC) is inhibited in the presence of anti-Fc?RIII blocking antibodies. This glycovariant of hPR1A3 enhances ADCC 10-fold relative to the parent unmodified antibody using either unfractionated peripheral blood mononuclear or natural killer (NK) cells and CEA-positive CRC cells as targets. NK cells are far more potent in eliciting ADCC than either freshly isolated monocytes or granulocytes. Flow cytometry and automated fluorescent microscopy have been used to show that both versions of hPR1A3 can induce antibody-dependent cellular phagocytosis (ADCP) by monocyte-derived macrophages. However, the glycovariant antibody did not mediate enhanced ADCP. This may be explained by the relatively low expression of Fc?RIIIa on cultured macrophages. In vivo studies show the efficacy of glycoengineered humanised IgG1 PR1A3 in significantly improving survival in a CRC metastatic murine model. Conclusion: The greatly enhanced in vitro ADCC activity of the glycoengineered version of hPR1A3 is likely to be clinically beneficial. PMID:19904275

  14. Detecting Lyme disease using antibody-functionalized carbon nanotubes

    NASA Astrophysics Data System (ADS)

    Dailey, Jennifer; Lerner, Mitchell; Goldsmith, Brett; Brisson, Dustin; Johnson, A. T. Charlie

    2011-03-01

    We combine antibodies for Lyme flagellar protein with carbon nanotube transistors to create an electronic sensor capable of definitive detection of Lyme disease. Over 35,000 cases of Lyme disease are reported in the United States each year, of which more than 23 percent are originally misdiagnosed. Rational design of the coupling of the biological system to the electronic system gives us a flexible sensor platform which we can apply to several biological systems. By coupling these antibodies to carbon nanotubes in particular, we allow for fast, sensitive, highly selective, electronic detection. Unlike antibody or biomarker detection, bacterial protein detection leads to positive identification of both early and late stage bacterial infections, and is easily expandable to environmental monitoring.

  15. Neutralising Antibodies against Ricin Toxin

    PubMed Central

    Prigent, Julie; Panigai, Laetitia; Lamourette, Patricia; Sauvaire, Didier; Devilliers, Karine; Plaisance, Marc; Volland, Hervé; Créminon, Christophe; Simon, Stéphanie

    2011-01-01

    The Centers for Disease Control and Prevention have listed the potential bioweapon ricin as a Category B Agent. Ricin is a so-called A/B toxin produced by plants and is one of the deadliest molecules known. It is easy to prepare and no curative treatment is available. An immunotherapeutic approach could be of interest to attenuate or neutralise the effects of the toxin. We sought to characterise neutralising monoclonal antibodies against ricin and to develop an effective therapy. For this purpose, mouse monoclonal antibodies (mAbs) were produced against the two chains of ricin toxin (RTA and RTB). Seven mAbs were selected for their capacity to neutralise the cytotoxic effects of ricin in vitro. Three of these, two anti-RTB (RB34 and RB37) and one anti-RTA (RA36), when used in combination improved neutralising capacity in vitro with an IC50 of 31 ng/ml. Passive administration of association of these three mixed mAbs (4.7 µg) protected mice from intranasal challenges with ricin (5 LD50). Among those three antibodies, anti-RTB antibodies protected mice more efficiently than the anti-RTA antibody. The combination of the three antibodies protected mice up to 7.5 hours after ricin challenge. The strong in vivo neutralising capacity of this three mAbs combination makes it potentially useful for immunotherapeutic purposes in the case of ricin poisoning or possibly for prevention. PMID:21633505

  16. Antibodies to watch in 2015.

    PubMed

    Reichert, Janice M

    2015-01-01

    The commercial pipeline of recombinant antibody therapeutics is robust and dynamic. As of early December 2014, a total of 6 such products (vedolizumab, siltuximab, ramucirumab, pembrolizumab, nivolumab, blinatumomab) were granted first marketing approvals in 2014. As discussed in this perspective on antibodies in late-stage development, the outlook for additional approvals, potentially still in 2014 and certainly in 2015, is excellent as marketing applications for 7 antibody therapeutics (secukinumab, evolocumab, mepolizumab, dinutuximab, nivolumab, blinatumomab, necitumumab) are undergoing a first regulatory review in the EU or US. Of the 39 novel mAbs currently in Phase 3 studies, a marketing application for one (alirocumab) may be submitted in late 2014, and marketing application submissions for at least 4 (reslizumab, ixekizumab, ocrelizumab, obiltoxaximab) are expected in 2015. Other 'antibodies to watch' are those in Phase 3 studies with estimated primary completion dates in late 2014 or 2015, which includes 13 for non-cancer indications (brodalumab, bimagrumab, bococizumab, MABp1, gevokizumab, dupilumab, sirukumab, sarilumab, tildrakizumab, guselkumab, epratuzumab, combination of actoxumab + bezlotoxumab, romosozumab) and 2 (racotumomab and clivatuzumab tetraxetan) undergoing evaluation as treatments for cancer. In addition to the novel antibody therapeutics mentioned, biosimilar infliximab and biosimilar trastuzumab are 'antibodies to watch' in 2015 because of their potential for entry into the US market and regulatory review, respectively. PMID:25484055

  17. Antibodies to watch in 2014.

    PubMed

    Reichert, Janice M

    2014-01-01

    Since 2010, mAbs has documented the biopharmaceutical industry's progress in transitioning antibody therapeutics to first Phase 3 clinical studies and regulatory review, and its success at gaining first marketing approvals for antibody-based products. This installment of the "Antibodies to watch" series outlines events anticipated to occur between December 2013 and the end of 2014, including first regulatory actions on marketing applications for vedolizumab, siltuximab, and ramucirumab, as well as the Fc fusion proteins Factor IX-Fc and Factor VIII-Fc; and the submission of first marketing applications for up to five therapeutics (secukinumab, ch14.18, onartuzumab, necitumumab, gevokizumab). Antibody therapeutics in Phase 3 studies are described, with an emphasis on those with study completion dates in 2014, including antibodies targeting interleukin-17a or the interleukin-17a receptor (secukinumab, ixekizumab, brodalumab), proprotein convertase subtilisin/kexin type 9 (alirocumab, evolocumab, bococizumab), and programmed death 1 receptor (lambrolizumab, nivolumab). Five antibodies with US Food and Drug Administration's Breakthrough Therapy designation (obinutuzumab, ofatumumab, lambrolizumab, bimagrumab, daratumumab) are also discussed. PMID:24284914

  18. Antibodies to watch in 2014

    PubMed Central

    Reichert, Janice M

    2014-01-01

    Since 2010, mAbs has documented the biopharmaceutical industry’s progress in transitioning antibody therapeutics to first Phase 3 clinical studies and regulatory review, and its success at gaining first marketing approvals for antibody-based products. This installment of the “Antibodies to watch” series outlines events anticipated to occur between December 2013 and the end of 2014, including first regulatory actions on marketing applications for vedolizumab, siltuximab, and ramucirumab, as well as the Fc fusion proteins Factor IX-Fc and Factor VIII-Fc; and the submission of first marketing applications for up to five therapeutics (secukinumab, ch14.18, onartuzumab, necitumumab, gevokizumab). Antibody therapeutics in Phase 3 studies are described, with an emphasis on those with study completion dates in 2014, including antibodies targeting interleukin-17a or the interleukin-17a receptor (secukinumab, ixekizumab, brodalumab), proprotein convertase subtilisin/kexin type 9 (alirocumab, evolocumab, bococizumab), and programmed death 1 receptor (lambrolizumab, nivolumab). Five antibodies with US Food and Drug Administration’s Breakthrough Therapy designation (obinutuzumab, ofatumumab, lambrolizumab, bimagrumab, daratumumab) are also discussed. PMID:24284914

  19. Increased levels of anti-glycan antibodies in patients with cystic fibrosis

    PubMed Central

    2011-01-01

    Background The prevalence of Crohn's disease (CD) is increased in patients with cystic fibrosis (CF). Anti-Saccharomyces cerevisiae antibodies (ASCA) have been suggested as a screening tool to detect CD in CF. Recently, several new anti-glycan antibodies have been reported in CD. Materials and methods The sera of 119 CF patients of various age groups were prospectively screened for ASCA type IgG (gASCA), anti-laminaribioside carbohydrate IgG antibodies (ALCA), anti-chitobioside carbohydrate IgA antibodies (ACCA), and anti-mannobioside carbohydrate IgG antibodies (AMCA). The frequency of these anti-glycan antibodies was then compared in patients with CD, ulcerative colitis, rheumatoid arthritis and healthy volunteers. Results A significant number of CF patients were positive for gASCA (51.3% [41.6-60.6]) and up to three other anti-glycan antibodies concurrently. Serum levels of anti-glycan antibodies in CF and CD were not related to parameters of inflammation. Despite the well-documented difference in clinical course between male and female CF patients no gender difference of anti-glycan antibodies was found. In contrast, there was a significant positive correlation between anti-glycan markers and age in CF patients. Conclusions Our findings demonstrate for the first time the increased frequency of a panel of anti-glycan antibodies in CF and provide a link between the presence of these serological biomarkers and patient's age. Anti-glycan antibody profiling may therefore become a valuable tool in the care of patients with CF. PMID:22024437

  20. Metabolic exploitation of the sialic acid biosynthetic pathway to generate site-specifically labeled antibodies.

    PubMed

    Rochefort, Matthew M; Girgis, Mark D; Ankeny, Jacob S; Tomlinson, James S

    2014-01-01

    Lack of a universal site-specific conjugation methodology for antibodies limits their potential to be developed as tumor-specific imaging agents or targeted therapeutics. A potential mechanism for site-specific conjugation involves utilization of the conserved N-glycosylation site in the CH2 domain. We sought to develop an antibody with an altered azido-sugar at this site whereby site-specific label could be added. The HB8059 hybridoma was cultured with peracetylated N-azidoacetlymannosamine (Ac4ManNAz). The resulting azido-sugar antibody was conjugated to phosphine-polyethylene glycol (PEG3)-biotin via a modified Staudinger reaction. Biochemical and functional characterization of the biotinylated antibody was performed. The azido-sugar antibody was also labeled with DyLight-650-Phosphine and injected into mice harboring pancreatic cancer xenografts. The tumors were dissected and imaged utilizing an IVIS fluorescent camera. The antibody was successfully produced in 100 ?M Ac4ManNAz. The biotinylated antibody demonstrated a 50 kDa heavy and 25 kDa light chain on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, but demonstrated a single band at 50 kDa on western blot. Treatment with a N-linked glycosidase extinguished the band. Flow cytometry demonstrated antigen-specific binding of CA19-9-positive cells and the antibody localized to the antigen-positive tumor in vivo. We successfully produced an antibody with an azido-sugar at the conserved CH2 glycosylation site. We were able to utilize this azide to label the antibody with biotin or fluorescent label and demonstrate that the label is added in a site-specific manner to the heavy chain, N-linked glycosylation site. Finally, we demonstrated functionality of our antibody for in vitro and in vivo targeting of pancreatic cancer cells. PMID:24150277

  1. Acute eosinophilic ascites: an unusual form of an unusual case.

    PubMed

    Kodan, Parul; Shetty, Meenakshi A; Pavan, M R; Kariappa, Ahalya; Mahabala, Chakrapani

    2015-01-01

    Eosinophilic gastroenteritis (EGE) is an uncommon disease characterised by eosinophilic infiltration in the gastrointestinal tract. EGE may involve more than one layer of the gastrointestinal tract. Clinical features depend on the layer and location which is involved. We report an unusual case of eosinophilic ascites associated with antinuclear antibody positivity, which is an unusual variety of serosal form of EGE. PMID:25315240

  2. Elimination mechanisms of therapeutic monoclonal antibodies.

    PubMed

    Tabrizi, Mohammad A; Tseng, Chih-Ming L; Roskos, Lorin K

    2006-01-01

    Targeted therapies using monoclonal antibodies have achieved important therapeutic applications in the treatment of various human diseases. Understanding the factors that impact the pharmacokinetics of monoclonal antibodies is of high importance for effective therapy. Many factors related to the target antigen, antibody and patients can affect antibody elimination. Evaluation of these factors will facilitate the understanding of the processes involved in antibody elimination. PMID:16478695

  3. Classification of protein profiles from antibody microarrays using heat and detergent treatment.

    PubMed

    Häggmark, Anna; Neiman, Maja; Drobin, Kimi; Zwahlen, Martin; Uhlén, Mathias; Nilsson, Peter; Schwenk, Jochen M

    2012-06-15

    Antibody microarrays offer new opportunities for exploring the proteome and to identify biomarker candidates in human serum and plasma. Here, we have investigated the effect of heat and detergents on an antibody-based suspension bead array (SBA) assay using polyclonal antibodies and biotinylated plasma samples. With protein profiles from more than 2300 antibodies generated in 384-plex antibody SBAs, three major classes of heat and detergent susceptibility could be described. The results show that washing of the beads with SDS (rather than Tween) after target binding lowered intensity levels of basically all profiles and that about 50% of the profiles appeared to be lowered to a similar extent by heating of the sample. About 33% of the profiles appeared to be insensitive to heat treatment while another 17% showed a positive influence of heat to yield elevated profiles. The results suggest that the classification of antibodies is driven by the molecular properties of the antibody-antigen interaction and can generally not be predicted based on protein class or Western blot data. The experimental scheme presented here can be used to systematically categorize antibodies and thereby combine antibodies with similar properties into targeted arrays for analysis of plasma and serum. PMID:22023822

  4. Current status of cancer immunodetection with radiolabeled human monoclonal antibodies.

    PubMed

    De Jager, R; Abdel-Nabi, H; Serafini, A; Pecking, A; Klein, J L; Hanna, M G

    1993-04-01

    The use of radiolabeled murine monoclonal antibodies (MoAbs) for cancer immunodetection has been limited by the development of human antimouse antibodies (HAMA). Human monoclonal antibodies do not elicit a significant human antihuman (HAHA) response. The generation and production of human monoclonal antibodies met with technical difficulties that resulted in delaying their clinical testing. Human monoclonal antibodies of all isotypes have been obtained. Most were immunoglobulin (Ig) M directed against intracellular antigens. Two antibodies, 16.88 (IgM) and 88BV59 (IgG3k), recognize different epitopes on a tumor-associated antigen, CTA 16.88, homologous to cytokeratins 8, 18, and 19. CTA 16.88 is expressed by most epithelial-derived tumors including carcinomas of the colon, pancreas, breast, ovary, and lung. The in vivo targeting by these antibodies is related to their localization in nonnecrotic areas of tumors. Repeated administration of 16.88 over 5 weeks to a cumulative dose of 1,000 mg did not elicit a HAHA response. Two of 53 patients developed a low titer of HAHA 1 to 3 months after a single administration of 88BV59. Planar imaging of colorectal cancer with Iodine-131 (131I)-16.88 was positive in two studies in 9 of 12 and 16 of 20 patients preselected by immunohistochemistry. Tumors less than 2 cm in diameter are usually not detected. The lack of immunogenicity and long tumor residence time (average = 17 days) makes 16.88 a good candidate for therapy. Radioimmunlymphoscintigraphy with indium-111 (111In)-LiLo-16.88 administered by an intramammary route was used in the presurgical staging of primary breast cancer. The negative predictive value of lymph node metastases for tumors less than 3 cm was 90.5%. Planar and single photon emission computed tomography imaging of colorectal carcinoma with technetium-99m (99mTc) 88BV59 was compared with computed tomography (CT) scan in 36 surgical patients. The antibody scan was more sensitive than the CT scan in detecting abdominal and pelvic tumors: 68% versus 40% (P < .05). The combination of antibody scan and CT scan was superior to CT scan alone: 80% versus 40% (P < .01). Lesions as small as 0.5 cm in diameter were detected by antibody scan. The CT scan appears superior to the antibody scan for liver metastases. Patients with a high serum titer of HAMA from previous exposure to murine antibodies were successfully imaged. Antibody scans obtained with 99mTc-88BV59 have imaging characteristics similar to murine antibody scans obtained with radiolabeled IgGs. The absence or weak immunogenicity of the human monoclonal antibodies makes them good candidates for radioimmunodetection and radioimmunotherapy. PMID:8511602

  5. PREDOMINANCE OF IgM ANTI-U1RNP ANTIBODIES IN PATIENTS WITH SYSTEMIC LUPUS ERYTHEMATOSUS

    Microsoft Academic Search

    P. G. VLACHOYIANNOPOULOS; A. GUIALIS; A. G. TZIOUFAS; H. M. MOUTSOPOULOS

    1996-01-01

    SUMMARY Anti-UlRNP antibodies occur in patients with mixed connective tissue disease (MCTD), systemic sclerosis (SSc), systemic lupus erythematosus (SLE) and other ill-defined connective tissue diseases. To associate the isotypes of anti-UlRNP antibodies with the diagnosis of the disease, namely SLE or MCTD, sequential sera of patients positive for anti-UlRNP antibodies by counterimmunoelectrophorcsis (CIE) (32 with SLE, 35 with MCTD) were

  6. A minimal model of peptide binding predicts ensemble properties of serum antibodies

    PubMed Central

    2012-01-01

    Background The importance of peptide microarrays as a tool for serological diagnostics has strongly increased over the last decade. However, interpretation of the binding signals is still hampered by our limited understanding of the technology. This is in particular true for arrays probed with antibody mixtures of unknown complexity, such as sera. To gain insight into how signals depend on peptide amino acid sequences, we probed random-sequence peptide microarrays with sera of healthy and infected mice. We analyzed the resulting antibody binding profiles with regression methods and formulated a minimal model to explain our findings. Results Multivariate regression analysis relating peptide sequence to measured signals led to the definition of amino acid-associated weights. Although these weights do not contain information on amino acid position, they predict up to 40-50% of the binding profiles' variation. Mathematical modeling shows that this position-independent ansatz is only adequate for highly diverse random antibody mixtures which are not dominated by a few antibodies. Experimental results suggest that sera from healthy individuals correspond to that case, in contrast to sera of infected ones. Conclusions Our results indicate that position-independent amino acid-associated weights predict linear epitope binding of antibody mixtures only if the mixture is random, highly diverse, and contains no dominant antibodies. The discovered ensemble property is an important step towards an understanding of peptide-array serum-antibody binding profiles. It has implications for both serological diagnostics and B cell epitope mapping. PMID:22353141

  7. Antibodies to myelin oligodendrocyte glycoprotein in bilateral and recurrent optic neuritis

    PubMed Central

    Ramanathan, Sudarshini; Reddel, Stephen W.; Henderson, Andrew; Parratt, John D.E.; Barnett, Michael; Gatt, Prudence N.; Merheb, Vera; Kumaran, Raani-Yogeeta Anusuiya; Pathmanandavel, Karrnan; Sinmaz, Nese; Ghadiri, Mahtab; Yiannikas, Con; Vucic, Steve; Stewart, Graeme; Bleasel, Andrew F.; Booth, David; Fung, Victor S.C.; Dale, Russell C.

    2014-01-01

    Objective: We examined a cohort of adults with aquaporin-4 (AQP4) antibody–negative neuromyelitis optica/neuromyelitis optica spectrum disorder (NMO/NMOSD) for antibodies to myelin oligodendrocyte glycoprotein (MOG). Methods: We performed a flow cytometry cell-based assay using live human lentivirus–transduced cells expressing full-length surface MOG. Serum was tested in 23 AQP4 antibody–negative NMO/NMOSD patients with bilateral and/or recurrent optic neuritis (BON, n = 11), longitudinally extensive transverse myelitis (LETM, n = 10), and sequential BON and LETM (n = 2), as well as in patients with multiple sclerosis (MS, n = 76) and controls (n = 52). Results: MOG antibodies were detected in 9/23 AQP4 antibody–negative patients with NMO/NMOSD, compared to 1/76 patients with MS and 0/52 controls (p < 0.001). MOG antibodies were detected in 8/11 patients with BON, 0/10 patients with LETM, and 1/2 patients with sequential BON and LETM. Six of 9 MOG antibody–positive patients had a relapsing course. MOG antibody–positive patients had prominent optic disc swelling and were more likely to have a rapid response to steroid therapy and relapse on steroid cessation than MOG antibody–negative patients (p = 0.034 and p = 0.029, respectively). While 8/9 MOG antibody–positive patients had good follow-up visual acuity, one experienced sustained visual impairment, 3 had retinal nerve fiber layer thinning, and one had residual spinal disability. Conclusions: MOG antibodies have a strong association with BON and may be a useful clinical biomarker. MOG antibody–associated BON is a relapsing disorder that is frequently steroid responsive and often steroid dependent. Failure to recognize the disorder early and institute immunotherapy promptly may be associated with sustained impairment. Classification of evidence: This study provides Class II evidence that MOG antibodies are associated with AQP4 antibody–negative BON (sensitivity 69%, 95% confidence interval [CI] 42%–87%; specificity 99%, 95% CI 93.7%–99.8%). PMID:25364774

  8. Characterization of thyroid function and antithyroid antibody tests among Saudis

    PubMed Central

    Jammah, Anwar A.; Alshehri, Anwar S.; Alrakhis, Afaf A.; Alhedaithy, Asma S.; Almadhi, Asma M.; Alkwai, Hala M.; Alhamad, Maram M.; Alzahrani, Saad H.

    2015-01-01

    Objective: To determine the reference intervals for thyroid function tests and the prevalence of thyroid autoimmunity in the Saudi population. Methods: A cross-sectional prospective study was conducted in King Khalid University Hospital, Riyadh, Saudi Arabia from January to June 2013. History and physical examination were obtained. Thyroid-stimulating hormone (TSH), free thyroxine (FT4), and free triiodothyronine (FT3) were measured by Electro-chemiluminescence Immunoassay system-assay. Anti-thyroperoxidase, and anti-thyroglobulin antibodies were measured using enzyme-linked immunosorbent-assay. Subjects with previous or a family history of thyroid disorders, those taking medications affecting thyroid function, pregnant or lactating women, and those with goiter were excluded. Individuals with positive antibodies were excluded from the final analysis of the TSH reference range, but were used to determine the prevalence of thyroid autoimmunity. Results: Out of 337 Saudi subjects initially screened, 132 (aged 13-60 years) were candidates for reference calculation, the mean±standard deviation, and (2.5th-97.5th) percentile of TSH (mIU/L) was 1.96±0.9 (0.59-4.37), for FT4 (pmol/L) was 15.47±1.83 (12.04-19.13), and for FT3 (pmol/L) was 5.22±0.7 (4.07-6.76). The TSH was higher in the antibodies positive group (2.5±1.17 mIU/L) compared with the negative one (1.96±0.9 mIU/L) (p<0.05). Finally, 26% of subjects were tested positive for antithyroid antibodies. Conclusion: The TSH reference range was similar to laboratory references. Thyroid antibodies were prevalent in Saudis, necessitating further work in larger scale studies. PMID:25987111

  9. Position indicator

    DOEpatents

    Tanner, David E. (Poway, CA)

    1981-01-01

    A nuclear reactor system is described in which a position indicator is provided for detecting and indicating the position of a movable element inside a pressure vessel. The movable element may be a valve element or similar device which moves about an axis. Light from a light source is transmitted from a source outside the pressure vessel to a first region inside the pressure vessel in alignment with the axis of the movable element. The light is redirected by a reflector prism to a second region displaced radially from the first region. The reflector prism moves in response to movement of the movable element about its axis such that the second region moves arcuately with respect to the first region. Sensors are arrayed in an arc corresponding to the arc of movement of the second region and signals are transmitted from the sensors to the exterior of the reactor vessel to provide indication of the position of the movable element.

  10. Pharmacokinetic characteristics of therapeutic antibodies.

    PubMed

    Wohlrab, Johannes

    2015-06-01

    Because of their high molecular weight and their highly hydrophilic character, therapeutic antibodies behave differently in terms of absorption, distribution and elimination compared to conventional drugs. Also, their pharmacokinetic profile varies significantly among individuals. After subcutaneous administration, antibodies are absorbed via the lymphatic system and become systemically bioavailable with some delay. The physicochemical properties of the molecules hinder their distribution from the bloodstream into the tissue. Elimination occurs by proteolysis in various organs (skin, muscle, liver), but mainly within the reticuloendothelial system. Also relevant is the elimination through target antigens (especially in the case of cell-bound target antigens) as well as a recycling process through binding to the neonatal Fc receptor that provides protection from lysosomal degradation. Depending on the immunogenicity of the therapeutic antibody and the individual immune response, neutralizing antibodies can develop. Pharmacokinetic conditions can be optimized by coadministration of, for example, methotrexate. Moreover, risk factors for the loss of immunological tolerance, such as on-demand therapy or elective switching of therapeutic antibodies, should be avoided. PMID:26018364

  11. A novel enzyme-linked immunosorbent assay (ELISA) for the detection of beryllium antibodies.

    PubMed

    Clarke, S M

    1991-03-01

    A novel immunological method has been developed for detecting antibodies (IgG molecules) specific to beryllium, a light metal used in industry and capable of causing chronic beryllium disease. Beryllium metal was vacuum deposited onto commercially available immunological microsticks, which were then exposed to test plasma containing the putative antibodies. Antigen-antibody complexes were located using a biotin-avidin amplification method. One employee diagnosed with chronic beryllium disease and one diagnosed as "sensitized" (lymphocyte transformation positive) exhibited antibody titers graphically and statistically different and higher than a pooled baseline control population. Plasma from these two employees (former beryllium workers) was used in four different approaches to validate the presence of beryllium antibodies. The assay proved to be reproducible. PMID:2010619

  12. Increased Prevalence of Brucella suis and Pseudorabies Virus Antibodies in Adults of an Isolated Feral Swine Population in Coastal South Carolina

    Microsoft Academic Search

    Cory S. Gresham; Charles A. Gresham; Michael J. Duffy; Charles T. Faulkner; Sharon Patton

    2002-01-01

    Two hundred twenty seven adult (.8 mo) feral swine (Sus scrofa) trapped from April through July 1999 at three locations on a coastal South Carolina (USA) peninsula with restricted ingress and egress were tested for Brucella suis and pseudorabies virus (PRV) an- tibodies. Approximately 44% of the animals tested positive for B. suis antibodies and 61% tested positive for antibodies

  13. Enzyme-Linked Immunosorbent Assay Using Glycoprotein and Monoclonal Antibody for Detecting Antibodies to Vesicular Stomatitis Virus Serotype New Jersey?

    PubMed Central

    Lee, Hyang-Sim; Heo, Eun-Jeong; Jeoung, Hye-Young; Ko, Hyo-Rim; Kweon, Chang-Hee; Youn, Hee-Jeong; Ko, Young-Joon

    2009-01-01

    In this study, an enzyme-linked immunosorbent assay (ELISA) using glycoprotein and a monoclonal antibody (MAb) was developed for the detection of antibodies to vesicular stomatitis virus (VSV) serotype New Jersey (NJ). The glycoprotein to be used as a diagnostic antigen was extracted from partially purified VSV-NJ, and a neutralizing MAb specific to VSV-NJ was incorporated to compete with antibodies in a blocking ELISA using glycoprotein (GP ELISA). The cutoff of the GP ELISA was set at 40% inhibition, which corresponded to a virus neutralization test (VNT) titer of 32. With this threshold, the GP ELISA exhibited 99.6% specificity for naïve sera (n = 3,005) from cattle (n = 1,040), pigs (n = 1,120), and horses (n = 845) from domestic farms. The GP ELISA did not cross-react with sera positive for foot-and-mouth disease virus, swine vesicular disease virus, or VSV serotype Indiana. The GP ELISA was more compatible with the VNT than was the nucleocapsid-based ELISA for VSV-NJ-positive sera (n = 19). Taken together, this GP ELISA could be a useful tool as an alternative to the VNT for detecting antibodies specific to VSV-NJ. PMID:19279165

  14. Changing Positions.

    ERIC Educational Resources Information Center

    LaRose, Albert J.

    1987-01-01

    Describes one search for a middle- or upper-management academic library position within the context of concerns expressed by respondents in a 1985 survey of 1,400 American Library Association (ALA) members by Bernstein and Leach. Factors examined include locale; availability and salary; staff relations; the application; the screening process; and…

  15. Bovine leukemia virus p24 antibodies reflect blood proviral load

    PubMed Central

    2012-01-01

    Background Bovine leukemia virus (BLV) is worldwide distributed and highly endemic in Argentina. Among the strategies to prevent BLV dissemination, a control plan based on the selective segregation of animals according to their proviral load (PVL) is promising for our dairy productive system. The objective of this work was to study the relationship between the blood PVL and the antibody level, in order to identify whether the individual humoral response, i.e. the anti-p24 or anti-whole-BLV particle, could be used as a marker of the blood level of infection and thus help to recruit animals that may pose a lower risk of dissemination under natural conditions. Results The prevalence of p24 antibodies on the 15 farms studied was over 66%. The prevalence of p24 and whole-BLV antibodies and PVL quantification were analyzed in all the samples (n?=?196) taken from herds T1 and 51. ROC analysis showed a higher AUC for p24 antibodies than whole-BLV antibodies (Zreactivity: 3.55, P?antibodies in herd T1. A significant positive correlation was observed between PVL values and p24 antibody reactivity in both farms (r T1?=?0.7, P?antibody reactors (n?=?311) of the other 13 farms. The mean of high PVL reactors within weak p24 reactors was 17.38% (SD?=?8.92). In 5/15 farms, the number of weak p24 reactors with high PVL was lower than 10%. Conclusions We found that the humoral response reflected the level of in vivo infection, and may therefore have useful epidemiological applications. Whereas the quantitative evaluation of blood proviral load using real-time PCR is expensive and technically demanding, the measurement of antibodies in blood by ELISA is relatively straightforward and could therefore constitute a cost-effective tool in a BLV control intervention strategy, especially in highly infected herds such as Argentinean dairy ones. PMID:23047073

  16. Monoclonal Antibodies for the Treatment of Cancer

    PubMed Central

    Shuptrine, Casey; Surana, Rishi; Weiner, Louis M.

    2012-01-01

    Over the past decade, the clinical utility of monoclonal antibodies has been realized and antibodies are now a mainstay for the treatment of cancer. Antibodies have the unique capacity to target and kill tumor cells while simultaneously activating immune effectors to kill tumor cells through the complement cascade or antibody-dependent cellular cytotoxicity (ADCC). This multifaceted mechanism of action combined with target specificity underlies the capacity of antibodies to elicit anti-tumor responses while minimizing the frequency and magnitude of adverse events. This review will focus on mechanisms of action, clinical applications and putative mechanisms of resistance to monoclonal antibody therapy in the context of cancer. PMID:22245472

  17. Monoclonal Antibodies in Diagnosis and Therapy

    NASA Astrophysics Data System (ADS)

    Waldmann, Thomas A.

    1991-06-01

    Monoclonal antibodies have been applied clinically to the diagnosis and therapy of an array of human disorders, including cancer and infectious diseases, and have been used for the modulation of immune responses. Effective therapy using unmodified monoclonal antibodies has, however, been elusive. Recently, monoclonal antibody-mediated therapy has been revolutionized by advances such as the definition of cell-surface structures on abnormal cells as targets for effective monoclonal antibody action, genetic engineering to create less immunogenic and more effective monoclonal antibodies, and the arming of such antibodies with toxins or radionuclides to enhance their effector function.

  18. B cell responses to HIV and the development of human monoclonal antibodies.

    PubMed Central

    Boyd, J E; James, K

    1992-01-01

    In this review B cell responses in HIV-infected individuals are summarized together with the techniques used to date to produce human monoclonals to HIV and the properties of these antibodies. Profound disturbances in B cell responses are apparent both in vivo and in vitro. While there is evidence in vivo of marked polyclonal B cell activation, primary and secondary antibody responses are impaired. Similarly these cells exhibit spontaneous immunoglobulin secretion upon in vitro culture but do not readily respond to B cell mitogens and recall antigens including HIV. Furthermore, certain of these defects can be reproduced in normal B cells in vitro by incubation with HIV or HIV coded peptides. Individuals infected with HIV develop antibodies to HIV structural proteins (e.g. p17, p24, gp41 and gp120) and regulatory proteins (e.g. vif, nef, RT). Autoantibodies against a number of immunologically important molecules are also frequently observed. The anti-HIV antibodies are predominantly of the IgG1 isotype and exhibit a variety of effects on the virus in vitro. To date, using conventional immortalization strategies, an appreciable number of human monoclonals to HIV have been developed. These have been specific for gp41, gp120 and gag with antibodies of the former specificity predominating. The majority of these antibodies have been of the IgG1 isotype. Only a small number of the antibodies neutralize virus in vitro and most of these react with gp120. The neutralizing antibodies recognize conformational and carbohydrate epitopes or epitopes in amino acid positions 306-322. The predominant epitopes recognized by the anti-gp41 antibodies were in amino acid positions 579-620 and 644-662. A high percentage (congruent to 25%) of these antibodies enhance viral growth in vitro. The problems relating to the production of human monoclonals to HIV are discussed together with strategies that could be used in the future. PMID:1572084

  19. Controlling chemical reactivity with antibodies

    SciTech Connect

    Hsieh, L.C.; Schultz, P.G. (Univ. of California, Berkeley (United States)); Yonkovich, S.; Kochersperger, L. (Affymax Research Inst., Palo Alto, CA (United States))

    1993-04-16

    The remarkable specificity of an antibody molecule has been used to accomplish highly selective functional group transformations not attainable by current chemical methods. An antibody raised against an amine-oxide hapten catalyzes the reduction of a diketone to a hydroxyketone with greater than 75:1 regioselectivity for one of two nearly equivalent ketone moieties. The antibody-catalyzed reaction is highly stereoselective, affording the hydroxyketone in high enantiomeric excess. Similarly, the reduction of ketones containing branched and aryl substituents, including the highly symmetrical 1-nitrophenyl-3-phenyl-2-propanone, was enantioselective. The simple strategy presented herein may find general applicability to the regio- and stereoselective reduction of a broad range of compounds. 18 refs., 2 figs., 1 tab.

  20. Molecular-specific urokinase antibodies

    NASA Technical Reports Server (NTRS)

    Atassi, M. Zouhair (Inventor); Morrison, Dennis R. (Inventor)

    2009-01-01

    Antibodies have been developed against the different molecular forms of urokinase using synthetic peptides as immunogens. The peptides were synthesized specifically to represent those regions of the urokinase molecules which are exposed in the three-dimensional configuration of the molecule and are uniquely homologous to urokinase. Antibodies are directed against the lysine 158-isoleucine 159 peptide bond which is cleaved during activation from the single-chain (ScuPA) form to the bioactive double chain (54 KDa and 33 KDa) forms of urokinase and against the lysine 135 lysine 136 bond that is cleaved in the process of removing the alpha-chain from the 54 KDa form to produce the 33 KDa form of urokinase. These antibodies enable the direct measurement of the different molecular forms of urokinase from small samples of conditioned medium harvested from cell cultures.

  1. Antibodies to watch in 2013

    PubMed Central

    Reichert, Janice M

    2013-01-01

    The transitions of antibody therapeutics to late-stage clinical development, regulatory review and the market are proceeding at a rapid pace in 2013. Since late 2012, two monoclonal antibody (mAb) therapeutics (itolizumab, trastuzumab emtansine) received their first approvals, first marketing applications for three mAbs (vedolizumab, ramucirumab, obinutuzumab) were submitted to regulatory agencies, and five mAbs (brodalumab, MABp1, moxetumomab pasudotox, tildrakizumab, rilotumumab) entered their first Phase 3 studies. The current total of commercially-sponsored antibody therapeutics undergoing evaluation in late-stage studies is 30. Recently announced study results for farletuzumab, naptumomab estafenatox, and tabalumab indicate that clinical endpoints were not met in some Phase 3 studies of these product candidates. PMID:23727858

  2. Entanglement model of antibody viscosity.

    PubMed

    Schmit, Jeremy D; He, Feng; Mishra, Shradha; Ketchem, Randal R; Woods, Christopher E; Kerwin, Bruce A

    2014-05-15

    Antibody solutions are typically much more viscous than solutions of globular proteins at equivalent volume fraction. Here we propose that this is due to molecular entanglements that are caused by the elongated shape and intrinsic flexibility of antibody molecules. We present a simple theory in which the antibodies are modeled as linear polymers that can grow via reversible bonds between the antigen binding domains. This mechanism explains the observation that relatively subtle changes to the interparticle interaction can lead to large changes in the viscosity. The theory explains the presence of distinct power law regimes in the concentration dependence of the viscosity as well as the correlation between the viscosity and the charge on the variable domain in our antistreptavidin IgG1 model system. PMID:24758234

  3. Characterization of antibodies directed against platelet cryptantigens detected during the immunological study of 356 consecutive patients with presumed autoimmune thrombocytopenia (AITP).

    PubMed

    Muñiz-Diaz, E; Madoz, P; Pujol-Moix, N; Pastoret, C; Arilla, M; Ibañez, M; Guanyabens, C; Domingo-Albós, A

    1995-09-01

    Cryptic antigens are detected by antibodies present in a wide spectrum of patients with or without thrombocytopenia, and even in healthy individuals. They are produced for unknown reasons and do not react with antigens of native platelets, but only with altered platelets. Cryptantigen antibodies may not only result in spuriously low platelet counts, but also in 'falsely' positive tests for platelet antibodies. We report our experience in the characterization of the different types of antibodies directed against cryptantigens of platelets: EDTA-dependent antibodies, PFA-dependent antibodies, EDTA-PFA-dependent antibodies and cold agglutinins. These antibodies were detected in the course of the serological study of 37 patients from a group of 356 (10%) whose blood was sent to our laboratory for platelet antibody testing. Pseudothrombocytopenia was diagnosed in 24 cases. Twenty-one of these showed EDTA-dependent or EDTA-PFA-dependent platelet agglutination and three were due to the presence of cold agglutinins. In 13 patients the thrombocytopenia was genuine. Eleven of these presented EDTA-dependent or EDTA-PFA-dependent antibodies in their serum and in the two remaining cases PFA-dependent antibodies were found. Cryptantigen antibodies were also detected in 9 out of 228 (4%) blood donors who were used as healthy controls in the platelet immunofluorescence test. In the light of the results obtained we put forward some guidelines to detect the presence of these antibodies and establish an accurate serological and clinical diagnosis of the autoimmune thrombocytopenias. PMID:8593522

  4. Antibody microarrays utilizing site-specific antibody-oligonucleotide conjugates.

    PubMed

    Wold, Erik D; McBride, Ryan; Axup, Jun Y; Kazane, Stephanie A; Smider, Vaughn V

    2015-05-20

    Protein arrays are typically made by random absorption of proteins to the array surface, potentially limiting the amount of properly oriented and functional molecules. We report the development of a DNA encoded antibody microarray utilizing site-specific antibody-oligonucleotide conjugates that can be used for cell immobilization as well as the detection of genes and proteins. This technology allows for the facile generation of antibody microarrays while circumventing many of the drawbacks of conventionally produced antibody arrays. We demonstrate that this method can be used to capture and detect SK-BR-3 cells (Her2+ breast cancer cells) at concentrations as low as 10(2) cells/mL (which is equivalent to 10 cells per 100 ?L array) without the use of microfluidics, which is 100- to 10(5)-fold more sensitive than comparable techniques. Additionally, the method was shown to be able to detect cells in a complex mixture, effectively immobilizing and specifically detecting Her2+ cells at a concentration of 10(2) SK-BR-3 cells/mL in 4 × 10(6) white blood cells/mL. Patients with a variety of cancers can have circulating tumor cell counts of between 1 and 10(3) cells/mL in whole blood, well within the range of this technology. PMID:25884500

  5. Antiganglioside antibody in patients with Guillain-Barré syndrome who show bulbar palsy as an initial symptom

    PubMed Central

    Koga, M.; Yuki, N.; Hirata, K.

    1999-01-01

    OBJECTIVES—To identify valuable antiganglioside antibodies that support the diagnosis of Guillain-Barré syndrome (GBS) and its variants in patients showing bulbar palsy as an initial symptom.?METHODS—Medical records of 602 patients with GBS or its variants were reviewed. Fifteen patients had bulbar palsy as an initial symptom. Serum antibodies against GM1, GM1b, GD1a, GalNAc-GD1a, GT1a, and GQ1b were examined in 13 of them.?RESULTS—Serum antiganglioside antibodies were positive in 11 (85%) patients. IgG anti-GT1a (n=8; 62%) and anti-GM1b (n=7; 54%) antibodies were often present, whereas all the patients had low or no anti-GM1 antibody activity. High anti-GD1a and anti-GQ1b IgG antibody titres were also present in some patients, but most had higher IgG antibody titres to GM1b or GT1a. All five patients with high IgG antibody titre to GM1b or GT1a only had had antecedent diarrhoea. Some patients with pharyngeal-cervical-brachial weakness (PCB) had IgG antibody to GT1a which did not cross react with GQ1b. Other patients with PCB had antibody to GT1a which cross reacted with GQ1b or antibody to GM1b, but anti-GM1b and anti-GT1a antibodies were not associated with the presence of bulbar palsy. All the patients who had no IgG antiganglioside antibodies recovered completely.?CONCLUSIONS—Measurement of serum IgG anti-GT1a and anti-GM1b antibodies gives helpful support for the diagnosis of GBS and its variants when there is early involvement of the oropharyngeal function independently of other neurological findings which appear as the illness progresses.?? PMID:10201426

  6. Mass screening for coeliac disease using antihuman transglutaminase antibody assay

    Microsoft Academic Search

    A Tommasini; V Kiren; V Baldas; D Santon; C Trevisiol; I Berti; E Neri; T Gerarduzzi; I Bruno; A Lenhardt; E Zamuner; A Spano?; S Crovella; S Martellossi; G Torre; D Sblattero; R Marzari; A Bradbury; G Tamburlini; A Ventura

    2004-01-01

    Aims: To determine coeliac disease prevalence by an anti-transglutaminase antibody assay in a large paediatric population; to evaluate acceptance of the screening programme, dietary compliance, and long term health effects.Methods: Cross-sectional survey of 3188 schoolchildren (aged 6–12) and prospective follow up of diagnosed cases. Main outcome measures were: prevalence of coeliac disease defined by intestinal biopsy or positivity to both

  7. A monoclonal antibody against a new differentiation antigen of thymocytes

    Microsoft Academic Search

    Charles L. Sidman; Luciana Forni; G K ohler; Jean Langhorne; Kirsten Fischer Lindahl

    1983-01-01

    B14-2-14 is a monoclonal cytotoxic IgM antibody which reacts with thymocytes of all mouse strains tested. The fraction of positive cells (by visual immunofluorescence) varies between strains from about 25-45% in A.CA to 65-85% in C57BL\\/6, and high levels are dominant in F1 hybrids. In the periphery, the antigen is found on a few percent of lymph node and not

  8. Glycine receptor antibodies in PERM and related syndromes: characteristics, clinical features and outcomes

    PubMed Central

    Carvajal-González, Alexander; Leite, M. Isabel; Waters, Patrick; Woodhall, Mark; Coutinho, Ester; Balint, Bettina; Lang, Bethan; Pettingill, Philippa; Carr, Aisling; Sheerin, Una-Marie; Press, Raomand; Lunn, Michael P.; Lim, Ming; Maddison, Paul; Meinck, H.-M.; Vandenberghe, Wim

    2014-01-01

    The clinical associations of glycine receptor antibodies have not yet been described fully. We identified prospectively 52 antibody-positive patients and collated their clinical features, investigations and immunotherapy responses. Serum glycine receptor antibody endpoint titres ranged from 1:20 to 1:60 000. In 11 paired samples, serum levels were higher than (n = 10) or equal to (n = 1) cerebrospinal fluid levels; there was intrathecal synthesis of glycine receptor antibodies in each of the six pairs available for detailed study. Four patients also had high glutamic acid decarboxylase antibodies (>1000 U/ml), and one had high voltage-gated potassium channel-complex antibody (2442 pM). Seven patients with very low titres (<1:50) and unknown or alternative diagnoses were excluded from further study. Three of the remaining 45 patients had newly-identified thymomas and one had a lymphoma. Thirty-three patients were classified as progressive encephalomyelitis with rigidity and myoclonus, and two as stiff person syndrome; five had a limbic encephalitis or epileptic encephalopathy, two had brainstem features mainly, two had demyelinating optic neuropathies and one had an unclear diagnosis. Four patients (9%) died during the acute disease, but most showed marked improvement with immunotherapies. At most recent follow-up, (2–7 years, median 3 years, since first antibody detection), the median modified Rankin scale scores (excluding the four deaths) decreased from 5 at maximal severity to 1 (P < 0.0001), but relapses have occurred in five patients and a proportion are on reducing steroids or other maintenance immunotherapies as well as symptomatic treatments. The glycine receptor antibodies activated complement on glycine receptor-transfected human embryonic kidney cells at room temperature, and caused internalization and lysosomal degradation of the glycine receptors at 37°C. Immunoglobulin G antibodies bound to rodent spinal cord and brainstem co-localizing with monoclonal antibodies to glycine receptor-?1. Ten glycine receptor antibody positive samples were also identified in a retrospective cohort of 56 patients with stiff person syndrome and related syndromes. Glycine receptor antibodies are strongly associated with spinal and brainstem disorders, and the majority of patients have progressive encephalomyelitis with rigidity and myoclonus. The antibodies demonstrate in vitro evidence of pathogenicity and the patients respond well to immunotherapies, contrasting with earlier studies of this syndrome, which indicated a poor prognosis. The presence of glycine receptor antibodies should help to identify a disease that responds to immunotherapies, but these treatments may need to be sustained, relapses can occur and maintenance immunosuppression may be required. PMID:24951641

  9. Evaluation of antibodies reactive with Campylobacter jejuni in Egyptian diarrhea patients.

    PubMed Central

    Ismail, T F; Wasfy, M O; Oyofo, B A; Mansour, M M; El-Berry, H M; Churilla, A M; Eldin, S S; Peruski, L F

    1997-01-01

    Serum and stool samples were collected from 128 individuals: 96 diarrhea patients and 32 apparently healthy controls. Stool specimens were cultured for enteric bacterial pathogens, while sera were screened by enzyme-linked immunosorbent assay for Campylobacter jejuni-reactive antibodies. Of 28 diarrhea patients who demonstrated C. jejuni-reactive antibodies (titers, > 100), 14 were culture positive for this organism. The 32 healthy controls showed significantly lower antibody titers (P < 0.05) with the exception of 10 subjects who were culture positive for C. jejuni and had reactive immunoglobulin M (IgM) (6 subjects) and IgG (7 subjects). IgA was not detected in those 10 individuals (asymptomatic). Avidity was expressed as the thiocyanate ion concentration required to inhibit 50% of the bound antibodies. The avidity was higher in symptomatic patients than asymptomatic healthy controls. IgG was less avid (0.92 M) compared to IgM (0.1 M) and IgA (1.1 M), with no correlation between antibody titer and avidity. However, the thiocyanate ion concentration required for the complete inhibition of IgG (5 M)-bound antibodies was higher than that of IgA (2 M) and IgM (3 M). This study also shows that C. jejuni antibodies were variably cross-reactive with Escherichia coli, Shigella flexneri, Shigella sonnei, and Neisseria meningitidis in addition to Campylobacter coli and Campylobacter rectus. PMID:9302201

  10. Isotype specific ELISAs to detect antibodies against swine vesicular disease virus and their use in epidemiology.

    PubMed Central

    Dekker, A.; van Hemert-Kluitenberg, F.; Baars, C.; Terpstra, C.

    2002-01-01

    Isotype specific ELISAs to detect antibodies against swine vesicular disease, which may help to estimate the moment of infection, were developed and validated on sera from pigs experimentally infected with four different isolates of swine vesicular disease virus. Virus specific IgM antibodies could be detected from days 3-49 and occasionally up to day 91 after infection. IgG1 antibodies were first detected at day 8 and IgG2 at day 11. IgA antibodies coincided with IgG1 antibodies, but antibody titres varied widely. From the results obtained with the sera from the experimentally infected pigs, we calculated the day at which 50% of the pigs had become positive (D50). A D50 of 5, 4, 12, 12 and 24 days was calculated, respectively, for the appearance of antibodies in the virus neutralization test, the IgM, total IgG, IgG1 and IgG2 ELISA. A D50 of 49 days was calculated for the disappearance of IgM antibodies. The isotype specific ELISAs proved to be valuable tools to study the epidemiology of the disease. PMID:12002546

  11. Fine specificities of anti-LM1 IgG antibodies in Guillain-Barré syndrome.

    PubMed

    Susuki, Keiichiro; Yuki, Nobuhiro; Hirata, Koichi; Kuwabara, Satoshi

    2002-03-30

    We investigated the prevalence of anti-LM1 IgG antibody and its fine specificity in Guillain-Barré syndrome (GBS). Anti-LM1 IgG and IgM antibodies from sera of 47 patients with GBS--19 with acute inflammatory demyelinating polyneuropathy (AIDP), 27 with acute motor axonal neuropathy (AMAN), and 1 with acute motor-sensory axonal neuropathy (AMSAN)--were tested. Anti-LM1 IgG antibody was detected in only one patient with AIDP, whereas it was present in seven with AMAN and in one with AMSAN. Sera from the eight IgG anti-LM1-positive patients with AMAN/AMSAN also had IgG activity against the gangliosides GM1, GM1b, GD1a, GalNAc-GD1a, GD1b, or GQ1b. Anti-LM1 IgG antibodies from the AMAN/AMSAN patients cross-reacted with other gangliosides, whereas IgG antibody from the AIDP patient was monospecific against LM1. Anti-LM1 IgG antibody therefore, cannot be a marker of AIDP. In addition, whether monospecific anti-LM1 IgG antibody is associated with AIDP remains to be concluded. Larger studies are needed to verify whether monospecific anti-LM1 IgG antibody could be a marker of AIDP. PMID:11897245

  12. Evaluation of fluorescent-antibody tests as a means of confirming infant botulism.

    PubMed Central

    Glasby, C; Hatheway, C L

    1984-01-01

    Fluorescent-antibody techniques were evaluated for confirming infant botulism. Seventy-seven stool specimens from suspected cases were examined. All 34 specimens containing viable Clostridium botulinum at time of study gave positive results (29 on direct smears and 34 on enrichments). Two false-positive reactions were observed. Images PMID:6394626

  13. [Research into the antibody detection technology of mink plasmacytosis and its current applications].

    PubMed

    Wan, Hongli; Feng, Erkai; Wu, Hongchao; Yang, Yanling; Ni, Jia; Chen, Lizhi

    2015-01-01

    Mink plasmacytosis, caused by Aleutian Mink Disease Virus (AMDV), poses a threat to the development of the animal fur industry. Neutralizing antibodies against AMDV may result in a persistent infection rather than providing protection for minks. To date,no specific methods to prevent or cure this disease have been developed. In order to eliminate mink plasmacytosis, antibody detection technology has been used globally as a dominant approach to screen for AMDV-positive minks. This paper introduces the classical technology, counterimmunoelectrophoresis and emerging technology in terms of AMDV antibody detection,and provides a glimpse into the future development of these technologies. PMID:25997336

  14. The detection of antibody against peste des petits ruminants virus in Sheep, Goats, Cattle and Buffaloes

    Microsoft Academic Search

    Haider Ali Khan; Muhammad Siddique; Sajjad-ur-Rahman; Muhammad Abubakar; Muhammad Ashraf

    2008-01-01

    Monoclonal antibody-based competitive ELISA (C-ELISA) has been used for the specific measurement of antibodies to peste des\\u000a petits ruminants (PPR) viruses in sheep, goats, cattle and Buffalo. Serum samples from sheep (n?=?232), goats (n?=?428), cattle\\u000a (n?=?43), buffalo (n?=?89) were tested. The animals had not been vaccinated against rinderpest or PPR. Findings suggested\\u000a that the sero-positive cases were significantly higher in

  15. Human Glioma-Mesenchymal Extracellular Matrix Antigen Defined by Monoclonal Antibody

    Microsoft Academic Search

    Mario A. Bourdon; Carol J. Wikstrand; Heinz Furthmayr; Thomas J. Matthews; Dareil D. Bigner

    1983-01-01

    The distribution and localization of a glioma-associated antigen defined by monoclonal antibody 81C6 has been examined using human cultured cell lines and tissues. Monoclonal antibody 81C6 was selected from a hybridoma fusion of spleen cells of mice immunized with the glial fibrillary acidic protein-positive human glioma cell line U-251 MG. Results of cell surface radioimmu- noassay and absorption analysis demonstrated

  16. Two-Component Direct Fluorescent-Antibody Assay for Rapid Identification of Bacillus anthracis

    Microsoft Academic Search

    Barun K. De; Sandra L. Bragg; Gary N. Sanden; Kathy E. Wilson; Lois A. Diem; Chung K. Marston; Alex R. Hoffmaster; Gwen A. Barnett; Robbin S. Weyant; Teresa G. Abshire; John W. Ezzell; Tanja Popovic

    A two-component direct fluorescent-antibody (DFA) assay, using fluorescein-labeled monoclonal antibodies specific to the Bacillus anthracis cell wall (CW-DFA) and capsule (CAP-DFA) antigens, was evaluated and validated for rapid identification of B. anthracis. We analyzed 230 B. anthracis isolates; 228 and 229 were positive by CW-DFA and CAP-DFA assays, respectively. We also tested 56 non-B. anthracis strains; 10 B. cereus and

  17. A monoclonal blocking-ELISA for detection of orthopoxvirus antibodies in feline sera

    Microsoft Academic Search

    Claus-Peter Czerny; Karin Wagner; Kurt Gessler; Anton Mayr; Oskar-Rüger Kaaden

    1996-01-01

    A double sandwich blocking-ELISA using a genus-specific neutralizing monoclonal antibody (MAb) against the vaccinia virus 32 kD adsorption protein (D8L open reading frame; ORF) was developed to detect orthopoxvirus (OPV) antibodies in sera. A collection of 2173 feline serum samples was examined in an epidemiological study. The blocking-ELISA revealed 44 (2%) sera with positive titres of 1:2-1:256. ELISA results were

  18. Impaired Antibody-dependent Cellular Cytotoxicity Mediated by Herceptin in Patients with Gastric Cancer1

    Microsoft Academic Search

    Koji Kono; Akihiro Takahashi; Fumiko Ichihara; Hidemitsu Sugai; Hideki Fujii; Yoshirou Matsumoto

    2002-01-01

    The humanized monoclonal antibody Herceptin, which specifically tar- gets HER-2\\/neu, exhibits growth inhibitory activity against HER-2\\/neu- overexpressing tumors and is approved for therapeutic use with proved survival benefit in patients with HER-2\\/neu-positive breast cancer. In the present study, we investigated whether Herceptin could affect the HER- 2\\/neu-overexpressing gastric cancer cells based on antibody-dependent cell-mediated cytotoxicity (ADCC) and compared immune effector

  19. Inhibitory effect of monoclonal antibodies on the growth of Babesia caballi

    Microsoft Academic Search

    Hiromi Ikadai; You Tamaki; Xuenan Xuan; Ikuo Igarashi; Satoru Kawai; Hideyuki Nagasawa; Kozo Fujisaki; Yutaka Toyoda; Naoyoshi Suzuki; Takeshi Mikami

    1999-01-01

    Monoclonal antibodies (mAbs) were produced against Babesia caballi (USDA strain) to define a species-specific antigen for use in diagnosis and vaccine development. Eight positive clones of B. caballi mAbs determined by indirect immunofluorescent antibody test were selected for purification and further characterisation. Confocal laser microscopy showed that the antigens recognised by the mAbs were located on the surface\\/cytoplasm, central part,

  20. Comparison of three serotests for the detection of pseudorabies antibodies in pigs.

    PubMed

    Goyal, S M; Joo, H S; Mourning, J R; McPherson, S W; Goyal, K

    1987-01-01

    The serum-neutralization test (SN), enzyme-linked immunosorbent assay (ELISA) and the radial immunodiffusion enzyme assay (RIDEA) were compared for the detection of pseudorabies (PRV) antibodies in swine sera. A total of 1285 serum samples were tested. All three tests were considered useful in determining the PRV antibody status of swine on a herd basis, but available evidence supports the continued use of SN as the definitive test because of possible false positive reactions associated with ELISA and RIDEA. PMID:2827946

  1. Is a Third-Trimester Antibody Screen in Rh+ Women Necessary?

    Microsoft Academic Search

    Jeffrey M. Rothenberg; Beata Weirermiller; Kelly Dirig; William W. Hurd; Jeanne Schilder; Alan Golichowski

    Objective: To determine the need for routine third-trimester antibody screening in Rh+ women. Study Design: An analytic case-control study. Methods: We identified Rh+ pregnant women who had received prenatal care and retrospectively analyzed their laboratory data. Patients were grouped into those with a positive third-trimester antibody screen (cases) and those with a negative third-trimester screen (controls). Because entry into a

  2. Computer-aided modeling of the binding sites of anti-fluorescein antibodies 

    E-print Network

    Harris, Jonathan S

    1996-01-01

    SPECTROSCOPY The mouse hybridomas producing monoclonal antibodies to the fluorescein hapten conjugated through the 4' position of the xanthene ring (4'-(aminomethyl) fluorescein) to the bovine serum albumin carrier protein were previously produced... of the subclasses has their own set of constant primers used for sequencing the variable domain. To determine the IgG subclass, goat antibodies to mouse immunoglobulins conjugated to biotin were incubated with 2 ml of each hybridoma supernatant for 30 minutes...

  3. Antibodies to islet 37k antigen, but not to glutamate decarboxylase, discriminate rapid progression to IDDM in endocrine autoimmunity.

    PubMed

    Christie, M R; Genovese, S; Cassidy, D; Bosi, E; Brown, T J; Lai, M; Bonifacio, E; Bottazzo, G F

    1994-10-01

    Apart from islet cell antibodies (ICAs), antibodies to glutamate decarboxylase (GAD), insulin autoantibodies (IAAs), and a novel islet antigen (37k antigen) are potential markers for insulin-dependent diabetes mellitus (IDDM). GAD is also an antigen in stiff-man syndrome (SMS), and both SMS and IDDM are associated with ICAs and autoimmunity to other endocrine organs. We investigated possible links between antibody responses to islet antigens with autoimmunity to other endocrine organs and determined which specific antibodies can identify individuals who progress to IDDM. Antibodies to GAD were detected in > or = 90% of both diabetic and nondiabetic patients with ICAs and other endocrine autoimmunity, in 59% of ICA-positive IDDM patients without endocrine autoimmunity, in all patients with SMS, but in only 1-3% of healthy (nondiabetic) and autoimmune disease control subjects. GAD antibody levels were increased in ICA-positive IDDM patients with polyendocrine autoimmunity compared with those without. In contrast, antibodies to 37k antigen were only detected in patients who developed acute-onset IDDM. IAAs were also associated with IDDM. Thus, certain factors enhance antibody responses to GAD in polyendocrine autoimmunity, but this does not necessarily lead to development of IDDM or SMS. Antibodies to 37k antigen are strongly associated with acute-onset IDDM and are useful serological markers for disease. PMID:7926297

  4. Detection of Campylobacter species using monoclonal antibodies

    NASA Astrophysics Data System (ADS)

    Young, Colin R.; Lee, Alice; Stanker, Larry H.

    1999-01-01

    A panel of species specific monoclonal antibodies were raised to Campylobacter coli, Campylobacter jejuni and Campylobacter lari. The isotypes, and cross-reactivity profiles of each monoclonal antibody against an extensive panel of micro- organisms, were determined.

  5. Development of a Human-Murine Chimeric Immunoglobulin M Antibody for Use in the Serological Detection of Human Flavivirus Antibodies

    Microsoft Academic Search

    Brett A. Thibodeaux; John T. Roehrig

    2009-01-01

    Current diagnosis of human flaviviral infections relies heavily on serological techniques such as the immu- noglobulin M (IgM) antibody capture enzyme-linked immunosorbent assay (MAC-ELISA). Broad application of this assay is hindered by a lack of standardized human positive-control sera that react with the wide variety of flaviviruses that can cause human disease, e.g., dengue virus (DENV), West Nile virus (WNV),

  6. Anti-insulin antibody test

    MedlinePLUS

    Insulin antibodies - serum; Insulin Ab test ... This test may be performed if you have or are at risk for type 1 diabetes . It also may ... different laboratories. Some labs use different measurements or test different samples. Talk to your doctor about the ...

  7. Anti-acetylcholine receptor antibodies.

    PubMed Central

    Vincent, A; Newsom Davis, J

    1980-01-01

    Early suggestions that a humoral factor might be implicated in the disorder of neuromuscular transmission in myasthenia gravis have been confirmed by the detection of anti-AChR antibody in 85-90% of the patients with generalised disease and in 75% of cases with restricted ocular myasthenia. Plasma exchange reveals that serum anti-AChR usually has an inverse relationship to muscle strength and present evidence indicates that patients responding to thymectomy and immunosuppressive durg treatment usually show a consistent decline in serum anti-AChR titres. The antibody is heterogeneous and can lead to a loss of muscle AChR by several mechanisms. Anti-AChR is produced in the thymus in relatively small amounts. Anti-AChR antibody synthesis by thymic lymphocytes and pokeweed stimulated peripheral lymphocytes in culture provides a means of studying the effect of different lymphocyte populations in vitro. Analysis of clinical, immunological and HLA antigen characteristics in MG suggest that more than one mechanism may underlie the breakdown in tolerance to AChR, leading to the production of anti-AChR antibodies. PMID:7400823

  8. Positioning apparatus

    DOEpatents

    Vogel, Max A. (Kennewick, WA); Alter, Paul (Richland, WA)

    1986-01-01

    An apparatus for precisely positioning materials test specimens within the optimum neutron flux path emerging from a neutron source located in a housing. The test specimens are retained in a holder mounted on the free end of a support pivotably mounted and suspended from a movable base plate. The support is gravity biased to urge the holder in a direction longitudinally of the flux path against the housing. Means are provided for moving the base plate in two directions to effect movement of the holder in two mutually perpendicular directions normal to the axis of the flux path.

  9. Therapeutic antibodies for autoimmunity and inflammation

    Microsoft Academic Search

    Andrew C. Chan; Paul J. Carter

    2010-01-01

    The development of therapeutic antibodies has evolved over the past decade into a mainstay of therapeutic options for patients with autoimmune and inflammatory diseases. Substantial advances in understanding the biology of human diseases have been made and tremendous benefit to patients has been gained with the first generation of therapeutic antibodies. The lessons learnt from these antibodies have provided the

  10. Monoclonal antibodies as neural cell surface markers

    Microsoft Academic Search

    O. K. Langley; M. S. Ghandour; G. Gombos; M. Hirn; C. Goridis

    1982-01-01

    A comparison is made of the immunohistochemistry at the ultrastructural level of three monoclonal antibodies directed against surface components of CNS cells. Hybridomas secreting these antibodies were obtained from two cell fusions of a rat myeloma cell line and immune splenocytes derived from rats immunized either with primary mouse brain cultured cells or membrane components. In cultures one antibody, anti-BSP-2

  11. Anti-DNA antibodies in SLE

    SciTech Connect

    Voss, E.W.

    1988-01-01

    This book contains 8 chapters. Some of the titles are: Anti-DNA Antibodies in SLE: Historical Perspective; Specificity of Anti-DNA Antibodies in Systemic Lupus Erythematosus; Monoclonial Autoimmune Anti-DNA Antibodies; and Structure--Function Analyses of Anti-DNA Autoantibodies.

  12. Immunohistochemical expression of p53 in animal tumors: a methodological study using four anti-human p53 antibodies.

    PubMed

    Albaric, O; Bret, L; Amardeihl, M; Delverdier, M

    2001-01-01

    Mutations in the p53 tumor suppressor gene are the most common genetic alterations in human cancers. These mutations usually lead to strongly enhanced protein stabilization and allow detection by immunohistochemistry. Two monoclonal (DO-7 and PAb-240) and two polyclonal (Ab-7 and CM-1) antibodies were evaluated by standard immunoperoxidase method in domestic animal tumors, chiefly squamous cell carcinomas (SCC), and osteosarcomas as positive controls. Immunoreactivity was detected in SCC of cattle, sheep, horse and cat as well as in feline actinic keratosis, with PAb-240 and CM-1 antibodies. One polyclonal antibody (Ab-7) did not give positive result at all, whereas DO-7 monoclonal antibody did not react in dogs and cats. Immunodetection of p53 protein is thus possible in all domestic species tested, especially with CM-1 and PAb-240 antibodies, and p53 alterations seem to occur early in carcinogenesis of feline SCC as in comparable human lesions. PMID:11193185

  13. Incidence of anti-HTLV-1 antibody in liver disease

    Microsoft Academic Search

    Hitoshi Nakano; Michio Sata; Kazuhiko Hino; Tomoki Aritaka; Naoto Maruyama; Teruko Hino; Hirohiko Abe; Kyuichi Tanikawa; Masashi Mizokami

    1992-01-01

    Summary  Positivity for serum anti-HTLV-1 antibody (anti-HTLV-1) in 171 patients with various chronic liver diseases and 22 asymptomatic\\u000a hepatitis B virus (HBV) carriers was compared with that of 200 healthy controls in the Chikugo district of Japan. The rate\\u000a of anti-HTLV-1 positivity in patients with liver disease was 8.1% (14\\/171) and was higher than that (3.5%: 7\\/200) in healthy\\u000a controls, but

  14. Anti-cardiolipin antibodies in sera from patients with periodontitis.

    PubMed

    Schenkein, H A; Berry, C R; Burmeister, J A; Brooks, C N; Barbour, S E; Best, A M; Tew, J G

    2003-11-01

    Antiphospholipid antibodies are commonly found in patients with systemic lupus erythematosus or the antiphospholipid syndrome, and a subset of such antibodies is associated with prothrombotic events such as stroke and with adverse pregnancy outcomes and fetal loss. We examined sera from 411 patients who were clinically characterized as to their periodontal disease status for serum levels of beta2-glycoprotein I-dependent anti-cardiolipin autoantibodies (anti-CL). The prevalence of patients with chronic periodontitis (CP) and generalized aggressive periodontitis (GAgP) positive for anti-CL (16.2% and 19.3%, respectively) was greater than that in healthy controls (NP) and localized aggressive periodontitis (LAgP) patients (6.8% and 3.2%). Patients with these autoantibodies demonstrated increased pocket depth and attachment loss compared with patients lacking the antibodies. Analysis of the data indicates that patients with generalized periodontitis have elevated levels of autoantibodies reactive with phospholipids. These antibodies could be involved in elevated risk for stroke, atherosclerosis, or pre-term birth in periodontitis patients. PMID:14578506

  15. Bouquet variety of antiphospholipid antibodies in recurrent pregnancy loss

    PubMed Central

    Karagyozova, Zhivka P.; Nikolova-Vlahova, Milena K.; Nikolov, Krasimir V.; Nikolov, Petar K.

    2014-01-01

    Seventy-six female patients with two or more recurrent pregnancy losses (RPL) during the 1st trimester were studied. Based on the results of the aCL and aB2GPI antibodies testing, patients were divided in two groups: 22 patients with RPL and elevated immunoglobulin (Ig) G/IgM aCL and/or aB2GPI [RPL + antiphospholipid syndrome (APS)] and 54 patients with RPL alone (without high antibodies). Immunoglobulin G aPS and IgG a-AnV in patients with RPL + APS were higher than in controls and IgG aPS were higher in RPL + APS than in RPL alone. Additionally IgG a-AnV and IgM aPE are higher in RPL alone than in controls. In 18/22 (81%) patients with RPL + APS and 29/54 (54%) patients with RPL alone, there were one or more positive antibodies: aPS, aPT, a-AnV or aPE. These results raise a question whether or not these antiphospholipid antibodies should be routinely tested in women with RPL and especially in the context of the so-called “seronegative APS”.

  16. Mesenchymal stem cell transplantation for diffuse alveolar hemorrhage in SLE

    Microsoft Academic Search

    Jun Liang; Fei Gu; Hong Wang; Bingzhu Hua; Yayi Hou; Songtao Shi; Liwei Lu; Lingyun Sun

    2010-01-01

    Background. A 19-year-old girl was diagnosed with systemic lupus erythematosus, based on findings of arthritis, malar rash, positive antinuclear antibody test and high levels of antibodies to double-stranded DNA. Two months after diagnosis, the patient presented with a sudden drop in blood hemoglobin level. Several days later, she developed bloody sputum, rapidly progressive dyspnea and hypoxemia. High-resolution CT showed diffuse

  17. Serum antibodies to Borrelia burgdorferi, Anaplasma phagocytophilum, and Babesia microti in recaptured white-footed mice.

    PubMed

    Magnarelli, Louis A; Williams, Scott C; Norris, Steven J; Fikrig, Erol

    2013-04-01

    A mark-release-recapture study was conducted during 2007 through 2010 in six, tick-infested sites in Connecticut, United States to measure changes in antibody titers for Borrelia burgdorferi sensu stricto, Anaplasma phagocytophilum, and Babesia microti in Peromyscus leucopus (white-footed mice). There was an overall recapture rate of 40%, but only four tagged mice were caught in ?2 yr. Sera from 561 mice were analyzed for total antibodies to B. burgdorferi and A. phagocytophilum by using whole-cell or recombinant (VlsE or protein 44) antigens in a solid-phase enzyme-linked immunosorbent assay or to whole-cell B. microti by indirect fluorescent antibody staining. Antibody prevalences were highly variable for B. burgdorferi (from 56% to 98%), A. phagocytophilum (from 11% to 85%), and B. microti (from 11% to 84%) depending on the site and time of sampling. Of 463 mice with antibodies, 206 (45%) had antibodies to all three pathogens. Changes in antibody status for some mice from negative to positive (117 seroconversions) or from positive to negative (55 reversions) were observed. Seroconversions were observed in 10.1% of 417 mice for B. burgdorferi, 18.0% of 306 mice for A. phagocytophilum, and 6.6% of 304 mice for B. microti; reversion rates were 5.3, 5.9, and 4.9%, respectively. Antibodies to all pathogens persisted in some mice over several weeks while, in others, there were marked declines in titration end points to negative status. The latter may indicate elimination of a certain pathogen, such as A. phagocytophilum, or that mouse immune systems ceased to produce antibodies despite an existing patent infection. PMID:23568904

  18. Identification of a linear epitope recognized by a monoclonal antibody directed to the heterogeneous nucleoriboprotein A2.

    PubMed

    Tronstrøm, Julie; Draborg, Anette Holck; Hansen, Paul Robert; Houen, Gunnar; Trier, Nicole Hartwig

    2014-01-01

    Rheumatoid arthritis (RA) is a chronic autoimmune disorder, characterized by progressive joint destruction and disability. Classical autoantibodies of RA are rheumatoid factors and citrulline antibodies. Patients positive for these autoantibodies are usually associated with a progressive disease course. A subgroup of RA patients does not express citrulline antibodies, instead are approximately 35% of these anti-citrulline-negative patients reported to express autoantibodies to the heterogeneous nucleoriboprotein A2, a ribonucleoprotein involved in RNA transport and processing also referred to as RA33. In the absence of citrulline antibodies, RA33 antibodies have been suggested to be associated with a milder disease course. In this study we screened the reactivity of a monoclonal antibody to RA33-derived peptides by modified enzyme-linked immunosorbent assays (ELISA). Terminally truncated resin-bound peptides were applied for determination of the functional epitope necessary for antibody recognition. In addition, screening of substituted peptides by modified ELISA identified amino acids necessary for antibody reactivity. A potential epitope was identified in the region 71-79 (PHSIDGRVV), where the amino acids Ser, Ile and Asp were found to be essential for antibody reactivity. These amino acids were found to contribute to the antibody-antigen interface through side-chain interactions, possibly in combination with a positively charged amino acid in position 77. Moreover, the amino acids in the N-terminal end (Pro and His) were found to contribute to the interface through backbone contributions. No notable reactivity was found with RA-positive patient sera, thus screening of RA33 antibodies does not seem to be a supplementary for the diagnosis of RA. PMID:23919377

  19. [Autoantibodies detected in acetylcholine receptor antibody-negative myasthenia gravis].

    PubMed

    Ohta, Rie; Motomura, Masakatsu

    2014-03-01

    Myasthenia gravis (MG) is caused by the failure of neuromuscular transmission mediated by pathogenic autoantibodies (Abs) against the acetylcholine receptor (AChR), muscle-specific receptor tyrosine kinase (MuSK), and unknown autoantibodies. The seropositivity rates for routine AChR binding Ab and MuSK Ab in MG are 85% and a few % for MG patients in Japan, respectively. The autoimmune target in the remaining patients is unknown. In 2001, Hoch et al. reported that a proportion of AChR-Ab-negative MG patients had serum IgG antibodies against MuSK, shedding new light on the pathogenesis of the disease. This idea has been recently supported by many clinical studies, including neonatal myasthenic syndrome and animal model studies. In 2011, autoantibodies against low-density lipoprotein receptor-related protein 4(Lrp4) were identified in Japanese MG patients and, thereafter, have been reported in Germany and the USA. We developed a simple technique termed Gaussia luciferase immunoprecipitation for detecting antibodies to Lrp4. As a result, nine generalized MG patients out of 300 lacking AChR Ab were found to be positive for Lrp4 antibodies. Thymoma was not observed in any of these patients. These antibodies inhibit the binding of Lrp4 to its ligand and are predominantly of the IgG1 subclass. In other reports of Lrp4 ab, Lrp4 ab-positive sera inhibited the agrin-induced aggregation of AChRs in cultured myotubes, suggesting a pathogenic role regarding the dysfunction of the neuromuscular endplate. These results indicate that Lrp4 is the third autoantigen in patients with MG, and anti-Lrp4 autoantibodies may be pathogenic. Further studies including neuromuscular junction biopsy are needed to clarify the pathomechanism of Lrp4 ab-positive MG. PMID:24800501

  20. Review on modeling anti-antibody responses to monoclonal antibodies.

    PubMed

    Gómez-Mantilla, José David; Trocóniz, Iñaki F; Parra-Guillén, Zinnia; Garrido, María J

    2014-10-01

    Monoclonal antibodies (mAbs) represent a therapeutic strategy that has been increasingly used in different diseases. mAbs are highly specific for their targets leading to induce specific effector functions. Despite their therapeutic benefits, the presence of immunogenic reactions is of growing concern. The immunogenicity identified as anti-drug antibodies (ADA) production due to the continuous administration of mAbs may affect the pharmacokinetics (PK) and/or the pharmacodynamics (PD) of mAbs administered to patients. Therefore, the immunogenicity and its clinical impact have been studied by several authors using PK modeling approaches. In this review, the authors try to present all those models under a unique theoretical mechanism-based framework incorporating the main considerations related to ADA formation, and how ADA may affect the efficacy or toxicity profile of some therapeutic biomolecules. PMID:25027160

  1. Twist Positivity

    NASA Astrophysics Data System (ADS)

    Jaffe, Arthur

    1999-11-01

    We study a heat kernel e-?H defined by a self-adjoint Hamiltonian H acting on a Hilbert space H, and a unitary representation U(g) of a symmetry group G of H, normalized so that the ground vector of H is invariant under U(g). The triple {H, U(g), H} defines a twisted partition function Zg and a twisted Gibbs expectation <·>g, Zg=TrH(U(g-1) e-?H) and <·>g=TrH(U(g-1)·e-?H)/TrH(U(g-1) e-?H). We say that {H, U(g), H} is twist positive if Zg>0. We say that {H, U(g), H} has a Feynman-Kac representation with a twist U(g), if one can construct a function space and a probability measure d?g on that space yielding (in the usual sense on products of coordinates) <·>g=?·d?g. Bosonic quantum mechanics provides a class of specific examples that we discuss. We also consider a complex bosonic quantum field ?(x) defined on a spatial s-torus Ts and with a translation-invariant Hamiltonian. This system has an (s+1)-parameter abelian twist group Ts×R that is twist positive and that has a Feynman-Kac representation. Given ??Ts and ??R, the corresponding paths are random fields ?(x, t) that satisfy the twist relation?(x, t+?)=ei???(x-?, t). We also utilize the twist symmetry to understand some properties of "zero-mass" limits, when the twist ?, ? lies in the complement of a set ?sing of singular twists.

  2. Development of Broadly Neutralizing Antibodies from Autologous Neutralizing Antibody Responses

    PubMed Central

    Derdeyn, Cynthia A.; Moore, Penny L.; Morris, Lynn

    2014-01-01

    Purpose of Review Detailed genetic and structural characterization has revealed that broadly neutralizing antibodies (bnAbs) against HIV-1 have unusually high levels of somatic hypermutation, long CDRH3 domains, and the ability to target one of four sites of vulnerability on the HIV-1 envelope (Env) glycoproteins. A current priority is to understand how bnAbs are generated during natural infection, and translate this information into immunogens that can elicit bnAb following vaccination. Recent Findings Strain-specific neutralizing antibodies (nAb) can acquire broad neutralizing capacity when the transmitted/founder Env or a specific Env variant is recognized by an unmutated rearranged germline that has the capacity to develop bnAb like features. This could be a relatively infrequent event, as only certain germlines appear to possess inherent features needed for bnAb activity. Furthermore, the glycosylation pattern and diversity of circulating HIV-1 Envs, as well as the state of the B cell compartment, may influence the activation and maturation of certain antibody lineages. Summary Collectively, studies over the last year suggest that the development of HIV-1 Env immunogens that bind and activate bnAb-like germlines is feasible. However, more information about the features of Env variants and the host factors that lead to breadth during natural infection is needed to elicit bnAbs through immunization. PMID:24662931

  3. Decreased serum antibody responses to recombinant pneumocystis antigens in HIV-infected and uninfected current smokers.

    PubMed

    Crothers, Kristina; Daly, Kieran R; Rimland, David; Goetz, Matthew Bidwell; Gibert, Cynthia L; Butt, Adeel A; Justice, Amy C; Djawe, Kpandja; Levin, Linda; Walzer, Peter D

    2011-03-01

    Serologic studies can provide important insights into the epidemiology and transmission of Pneumocystis jirovecii. Exposure to P. jirovecii can be assessed by serum antibody responses to recombinant antigens from the major surface glycoprotein (MsgC), although factors that influence the magnitude of the antibody response are incompletely understood. We determined the magnitudes of antibody responses to P. jirovecii in comparison to adenovirus and respiratory syncytial virus (RSV) in HIV-infected and uninfected patients and identified predictors associated with the magnitude of the response. We performed a cross-sectional analysis using serum samples and data from 153 HIV-positive and 92 HIV-negative subjects enrolled in a feasibility study of the Veterans Aging Cohort 5 Site Study (VACS 5). Antibodies were measured using an enzyme-linked immunosorbent assay (ELISA). Independent predictors of antibody responses were determined using multivariate Tobit regression models. The results showed that serum antibody responses to P. jirovecii MsgC fragments were significantly and independently decreased in current smokers. Antibodies to P. jirovecii also tended to be lower with chronic obstructive pulmonary disease (COPD), hazardous alcohol use, injection drug use, and HIV infection, although these results were not statistically significant. These results were specific to P. jirovecii and did not correlate with adenovirus. Antibody responses to RSV were in the inverse direction. Thus, current smoking was independently associated with decreased P. jirovecii antibody responses. Whether smoking exerts an immunosuppressive effect that affects the P. jirovecii antibody response, colonization, or subsequent risk for disease is unclear; prospective, longitudinal studies are needed to evaluate these findings further. PMID:21191078

  4. Anti-class II antibodies in AIDS patients and AIDS-risk groups.

    PubMed Central

    de la Barrera, S; Fainboim, L; Lugo, S; Picchio, G R; Muchinik, G R; de Bracco, M M

    1987-01-01

    The specificity of anti-lymphocyte antibodies was evaluated in AIDS patients and in individuals at risk of AIDS [R-AIDS: male homosexuals (Ho) and haemophiliacs (He)]. Antibodies capable of inducing antibody-dependent cell-mediated cytotoxicity (ADCC) against non-T cells and lymphoblastoid cell lines (P3HR-1K and Raji) were detected in AIDS patients and in R-AIDS with positive or negative human immune deficiency virus (HIV) serology. Anti-class II antigen specificity was revealed by experiments in which class II antigens on target cells were blocked with monoclonal anti-class II antibody (DA6,231) and the cytotoxic reaction induced by patient's sera was abolished. In contrast, ADCC was not impaired by preincubating the target cells with anti-class I monoclonal antibody (W6/32). Prevalence of antibodies to non-T cells was confirmed by standard C-mediated microlymphocytotoxicity. However, with this technique anti-T lymphocyte cytotoxicity was also observed in three AIDS patients with haemophilia. R-AIDS peripheral blood mononuclear cells (PBMC) were also cytotoxic against autologous non-T cells, and lysis was slightly increased by sensitization of the target cells with autologous serum. In addition to ADCC and C-mediated cytotoxicity, the specificity of anti-lymphocyte antibodies was assayed by their ability to interfere the binding of fluorescein-labelled anti-class II (HLA-DR) and anti-class I (W6/32) monoclonal antibodies to PBMC, non-T cells, P3HR-1K and Raji. Anti-class II specificity was confirmed, and antibody titres tended to be higher in Ho than in He R-AIDS, using non-T cells and Raji as targets. Higher titres of anti-class II antibodies in the Ho group could play a role in the different susceptibility of HIV-infected Ho when compared to HIV (+) He to develop AIDS. PMID:3501399

  5. Mechanistic Insights into the Neutralization of Cytotoxic Abrin by the Monoclonal Antibody D6F10

    PubMed Central

    Bagaria, Shradha; Ponnalagu, Devasena; Bisht, Shveta; Karande, Anjali A.

    2013-01-01

    Abrin, an A/B toxin obtained from the Abrus precatorius plant is extremely toxic and a potential bio-warfare agent. Till date there is no antidote or vaccine available against this toxin. The only known neutralizing monoclonal antibody against abrin, namely D6F10, has been shown to rescue the toxicity of abrin in cells as well as in mice. The present study focuses on mapping the epitopic region to understand the mechanism of neutralization of abrin by the antibody D6F10. Truncation and mutational analysis of abrin A chain revealed that the amino acids 74–123 of abrin A chain contain the core epitope and the residues Thr112, Gly114 and Arg118 are crucial for binding of the antibody. In silico analysis of the position of the mapped epitope indicated that it is present close to the active site cleft of abrin A chain. Thus, binding of the antibody near the active site blocks the enzymatic activity of abrin A chain, thereby rescuing inhibition of protein synthesis by the toxin in vitro. At 1?10 molar concentration of abrin:antibody, the antibody D6F10 rescued cells from abrin-mediated inhibition of protein synthesis but did not prevent cell attachment of abrin. Further, internalization of the antibody bound to abrin was observed in cells by confocal microscopy. This is a novel finding which suggests that the antibody might function intracellularly and possibly explains the rescue of abrin’s toxicity by the antibody in whole cells and animals. To our knowledge, this study is the first report on a neutralizing epitope for abrin and provides mechanistic insights into the poorly understood mode of action of anti-A chain antibodies against several toxins including ricin. PMID:23922965

  6. Anti-HLA antibody profile of Turkish patients with end-stage renal disease.

    PubMed

    Karahan, G E; Seyhun, Y; Oguz, F; Kekik, C; Onal, E; Caliskan, Y; Bakkaloglu, H; Yazici, H; Turkmen, A; Aydin, A E; Sever, M S; Eldegez, U; Carin, M N

    2009-11-01

    Exposure to human leukocyte antigens (HLA) via blood transfusions, pregnancies, and previous transplantations can result in anti-HLA antibody production. The presence of anti-HLA antibodies in recipient sera before transplantation is an important risk factor. To demonstrate the anti-HLA antibody status of Turkish end-stage renal disease (ESRD) patients, 674 patients (mean age, 40.35 +/- 13.15 years; female/male, 328/346) were enrolled into the study. Anti-HLA antibody screening and identification tests were performed using an enzyme-linked immunosorbent assay (ELISA) method. The panel-reactive antibody (PRA)-negative group consisted of 564 (83.6%) and the PRA-positive group consisted of 110 (17.3%) patients. Of the 110 (17.3%) PRA-positive patients, 43 (6.4%) were class I (+) and class II (-); 19 (2.8%) were class I (-) and class II (+); 48 (7.1%) were both class I and II (+). The most frequent antibodies were directed against the A2 crossreactive group (CREG) and the A10 CREG with less frequent reactions against the B7 CREG, indicating antibodies to both frequent (members of A2 CREG) and relatively rare (members of A10 CREG and B7 CREG antigens). These data also suggested that some antibodies occur at greater than expected frequency because of shared epitopes. Our findings confirmed the significant correlation between female gender, pregnancy, failed graft history, long dialysis duration, and blood transfusions with PRA positivity (P < .05). PMID:19917361

  7. Engineered antibodies accelerating drug discovery and development.

    PubMed

    Zhu, Z

    2001-12-01

    This meeting, organized by the Strategic Research Institute, reviewed advances in both engineering technologies and clinical development of antibody products. A panel of speakers, mainly from the biotechnology and pharmaceutical industries, covered several important issues in the development of antibody therapeutics, including new display technologies, human antibody generation, high-level expression of antibodies in both mammalian cells and in transgenic plants, and the latest data from ongoing clinical trials of antibody products. In the case of human antibody generation, Abgenix Inc's XenoMax technology combines the powers of the transgenic XenoMouse and in vitro B cell culture and selection to enable the robust identification of antibodies with rare properties and very high affinity. MAbstract from Crucell NV provides a unique approach to the identification of human antibodies directed against tumor-specific glycosylation variants and activation epitopes of otherwise normal cellular molecules. Regarding antibody production, the use of transgenic plants is gaining more interest among biopharmaceutical industrials. Promising results were also reported on several antibody-based therapeutic, including antibody-drug conjugates and prodrugs. PMID:15931562

  8. Antibody dissociation rates are predictive of neutralizing antibody (NAb) course: a comparison of interferon beta-1b-treated Multiple Sclerosis (MS) patients with transient versus sustained NAbs.

    PubMed

    Gibbs, Ebrima; Karim, Mohammad Ehsanul; Oger, Joel

    2015-03-01

    A proportion of multiple sclerosis (MS) patients treated with interferon-? (IFN?) develop neutralizing antibodies (NAbs), which can reduce therapeutic efficacy. In the Betaseron/Betaferon in Newly Emerging MS for Initial Treatment (BENEFIT) study, 88/277 patients developed NAbs, 48 having transient positivity and 29 having sustained positivity. This study aimed to investigate the antibody binding characteristics of serial sera in a subset of these two patient groups. Using Biacore™, a surface plasmon resonance-based technology that monitors biomolecular interactions in real time, we immobilized pure IFN?-1b and analyzed antibody binding responses and dissociation rates of these sera. NAb titers correlated directly with binding responses and inversely with dissociation rates, and sera from sustained NAb patients demonstrated significantly higher binding responses and slower dissociation rates than sera from transient NAb patients. Thus, transient and sustained NAbs are quantitatively and qualitatively different, and interestingly, binding responses and dissociation rates at month 12 could predict the NAb course. PMID:25543089

  9. Validation of a monoclonal antibody-based ELISA to detect antibodies directed against swine vesicular disease virus.

    PubMed

    Chénard, G; Bloemraad, M; Kramps, J A; Terpstra, C; Dekker, A

    1998-11-01

    A simple, rapid and sensitive competitive monoclonal antibody-based ELISA for the detection of antibodies directed against swine vesicular disease virus (SVDV) was developed. The ELISA was validated using field sera originating from SVDV-infected and non-infected Dutch pig herds, reference sera obtained from the Community Reference Laboratory for Swine Vesicular Disease at the Institute for Animal Health, Pirbright Laboratory, UK, and sera from animals infected experimentally. When testing 4277 sera originating from non-infected Dutch pig herds and collected as part of the national screening program, this ELISA had only 0.6% false positive results, whereas approximately 2% of false positive results were obtained with a conventional blocking ELISA used until recently. A sensitivity relative to the virus neutralisation test of > 97% was achieved when testing sera collected from Dutch pig farms where an outbreak of SVDV had occurred. All international reference sera scored consistently correct. Sera collected sequentially from pigs experimentally infected with SVDV isolates representing all currently recognized antigenic groups, were scored positive slightly earlier by the ELISA compared to the virus neutralisation test. This monoclonal antibody-based competitive ELISA for SVDV antibodies designated the Ceditest ELISA for SVDV-Ab, is as sensitive but more specific than the ELISA used until recently. Because sera are tested at a single dilution (1:5), incubations are carried out at room temperature and test results are available within 3 h, this ELISA is simple, easy to automate and therefore very suitable for screening large numbers of serum samples. PMID:9820579

  10. The functional repertoire of rabbit antibodies and antibody discovery via next-generation sequencing

    PubMed Central

    Kodangattil, Sreekumar; Huard, Christine; Ross, Cindy; Li, Jian; Gao, Huilan; Mascioni, Alessandro; Hodawadekar, Santosh; Naik, Snehal; Min-debartolo, Jessica; Visintin, Alberto; Almagro, Juan C

    2014-01-01

    To gain insight into the functional antibody repertoire of rabbits, the VH and VL repertoires of bone marrow (BM) and spleen (SP) of a naïve New Zealand White rabbit (NZW; Oryctolagus cuniculus) and that of lymphocytes collected from a NZW rabbit immunized (IM) with a 16-mer peptide were deep-sequenced. Two closely related genes, IGHV1S40 (VH1a3) and IGHV1S45 (VH4), were found to dominate (~90%) the VH repertoire of BM and SP, whereas, IGHV1S69 (VH1a1) contributed significantly (~40%) to IM. BM and SP antibodies recombined predominantly with IGHJ4. A significant proportion (~30%) of IM sequences recombined with IGHJ2. The VK repertoire was encoded by nine IGKV genes recombined with one IGKJ gene, IGKJ1. No significant bias in the VK repertoire of the BM, SP and IM samples was observed. The complementarity-determining region (CDR)-H3 and -L3 length distributions were similar in the three samples following a Gaussian curve with average length of 12.2 ± 2.4 and 11.1 ± 1.1 amino acids, respectively. The amino acid composition of the predominant CDR-H3 and -L3 loop lengths was similar to that of humans and mice, rich in Tyr, Gly, Ser and, in some specific positions, Asp. The average number of mutations along the IGHV/KV genes was similar in BM, SP and IM; close to 12 and 15 mutations for VH and VL, respectively. A monoclonal antibody specific for the peptide used as immunogen was obtained from the IM rabbit. The CDR-H3 sequence was found in 1,559 of 61,728 (2.5%) sequences, at position 10, in the rank order of the CDR-H3 frequencies. The CDR-L3 was found in 24 of 11,215 (0.2%) sequences, ranking 102. No match was found in the BM and SP samples, indicating positive selection for the hybridoma sequence. Altogether, these findings lay foundations for engineering of rabbit V regions to enhance their potential as therapeutics, i.e., design of strategies for selection of specific rabbit V regions from NGS data mining, humanization and design of libraries for affinity maturation campaigns. PMID:24481222

  11. Antiphospholipid antibody effects on monocytes

    Microsoft Academic Search

    Alisa S. Wolberg

    2007-01-01

    Although the presence of autoantibodies is known to increase the risk of thrombosis in the antiphospholipid syndrome, the\\u000a mechanism by which these antibodies exert their effects is poorly understood. Several studies suggest that autoantibody-mediated\\u000a dysregulation of monocytes is one pathobiologic mechanism of this disease. Recent studies have focused on extra-and intracellular\\u000a interactions involved in monocyte activation and expression of procoagulant

  12. Cytokine-neutralizing therapeutic antibodies

    Microsoft Academic Search

    Amanda Suitters; Roly Foulkes

    \\u000a Monoclonal antibodies (Mabs) offer the potential as useful therapeutic agents because of their high affinity and selectivity\\u000a for the target antigen. The standard approach in making Mabs has been to immunise rodents, usually mice, with the desired\\u000a human protein and generate a mouse anti-human IgG through hybridoma technology. Indeed mouse Mabs are used clinically in acute\\u000a diseases, such as the

  13. Therapeutic monoclonal antibodies in ophthalmology

    Microsoft Academic Search

    Eduardo B. Rodrigues; Michel E. Farah; Maurício Maia; Fernando M. Penha; Caio Regatieri; Gustavo B. Melo; Marcelo M. Pinheiro; Carlos R. Zanetti

    2009-01-01

    Monoclonal antibodies (mAbs) can be used therapeutically by binding to molecular targets with high specificity. Therefore, they have excellent therapeutic applications in ophthalmology. This manuscript presents four aspects of the therapeutic use of mAbs in ophthalmology: the scientific rationale, the unique characteristics of selected mAbs, the current state-of-the-art application, and relevant therapeutic mAbs for future applications in ophthalmology. We identified

  14. Bispecific Antibodies for Diagnostic Applications

    Microsoft Academic Search

    Archana Parashar; Susmita Sarkar; Advaita Ganguly; Sai Kiran Sharma; Mavanur R. Suresh

    \\u000a Bispecific monoclonal antibodies (BsMAb) are unique engineered macromolecules that have two different pre-determined binding\\u000a specificities. Their ability to simultaneously bind to a specific antigen and a given detection moiety enables them to function\\u000a as excellent bifunctional immunoprobes in diagnostic assays. BsMAb are being exploited for the development of simple, rapid,\\u000a and highly sensitive immunoassays for diagnosis of bacterial and viral

  15. Evaluation of recombinant vaccinia virus--measles vaccines in infant rhesus macaques with preexisting measles antibody.

    PubMed

    Zhu, Y d; Rota, P; Wyatt, L; Tamin, A; Rozenblatt, S; Lerche, N; Moss, B; Bellini, W; McChesney, M

    2000-10-10

    Immunization of newborn infants with standard measles vaccines is not effective because of the presence of maternal antibody. In this study, newborn rhesus macaques were immunized with recombinant vaccinia viruses expressing measles virus hemagglutinin (H) and fusion (F) proteins, using the replication-competent WR strain of vaccinia virus or the replication-defective MVA strain. The infants were boosted at 2 months and then challenged intranasally with measles virus at 5 months of age. Some of the newborn monkeys received measles immune globulin (MIG) prior to the first immunization, and these infants were compared to additional infants that had maternal measles-neutralizing antibody. In the absence of measles antibody, vaccination with either vector induced neutralizing antibody, cytotoxic T cell (CTL) responses to measles virus and protection from systemic measles infection and skin rash. The infants vaccinated with the MVA vector developed lower measles-neutralizing antibody titers than those vaccinated with the WR vector, and they sustained a transient measles viremia upon challenge. Either maternal antibody or passively transferred MIG blocked the humoral response to vaccination with both WR and MVA, and the frequency of positive CTL responses was reduced. Despite this inhibition of vaccine-induced immunity, there was a reduction in peak viral loads and skin rash after measles virus challenge in many of the infants with preexisting measles antibody. Therefore, vaccination using recombinant vectors such as poxviruses may be able to prevent the severe disease that often accompanies measles in infants. PMID:11022008

  16. Serosurvey for antibody to deerpox virus in five cervid species in Oregon, USA.

    PubMed

    Jin, Ling; McKay, Alison; Green, Richard; Xu, Linzhi; Bildfell, Rob

    2013-01-01

    Five cervid species in Oregon, USA were tested with a serum neutralization assay for antibody to deerpox virus (DPV). None of the 50 elk (Cervus elaphus ssp. roosevelti and nelsonii) had detectable antibody. Prevalence of antibody to DPV in the remaining species was: 52% (n=55) in black-tailed deer (Odocoileus hemionus columbianus), 32% (n= 59) in mule deer (O. hemionus hemionus), and 36% (n=50) in Columbian white-tailed deer (O. virginianus leucurus), with an overall antibody prevalence of 40.2% (n=164) for Odocoileus spp. Antibody-positive animals were identified throughout the state with no statistically significant differences among geographic regions. No statistically significant gender or age-related differences in antibody prevalence were demonstrated at either the genus or species level. This serosurvey indicates that exposure to DPV is common in Odocoileus populations in Oregon. Given the low rates of observed DPV-related disease, this high antibody prevalence suggests a pathogen of low virulence. PMID:23307387

  17. An Anti-Ubiquitin Antibody Response in Transitional Cell Carcinoma of the Urinary Bladder

    PubMed Central

    Ardelt, Peter U.; Ebbing, Jan; Adams, Fabian; Reiss, Cora; Arap, Wadih; Pasqualini, Renata; Bachmann, Alexander; Wetterauer, Ulrich; Riedmiller, Hubertus; Kneitz, Burkhard

    2015-01-01

    Background To use combinatorial epitope mapping (“fingerprinting”) of the antibody response to identify targets of the humoral immune response in patients with transitional cell carcinoma (TCC) of the bladder. Methods A combinatorial random peptide library was screened on the circulating pool of immunoglobulins purified from an index patient with a high risk TCC (pTa high grade plus carcinoma in situ) to identify corresponding target antigens. A patient cohort was investigated for antibody titers against ubiquitin. Results We selected, isolated, and validated an immunogenic peptide motif from ubiquitin as a dominant epitope of the humoral response. Patients with TCC had significantly higher antibody titers against ubiquitin than healthy donors (p<0.007), prostate cancer patients (p<0.0007), and all patients without TCC taken together (p<0.0001). Titers from superficial tumors were not significantly different from muscle invasive tumors (p = 0.0929). For antibody response against ubiquitin, sensitivity for detection of TCC was 0.44, specificity 0.96, positive predictive value 0.96 and negative predictive value 0.41. No significant titer changes were observed during the standard BCG induction immunotherapy. Conclusions This is the first report to demonstrate an anti-ubiquitin antibody response in patients with TCC. Although sensitivity of antibody production was low, a high specificity and positive predictive value make ubiquitin an interesting candidate for further diagnostic and possibly immune modulating studies. PMID:25742283

  18. PREVALENCE OF ANTIBODIES AGAINST INFLUENZA VIRUS IN NON-VACCINATED EQUINES FROM THE BRAZILIAN PANTANAL

    PubMed Central

    Silva, Lucas Gaíva E; Borges, Alice Mamede Costa Marques; Villalobos, Eliana Monteforte Cassaro; Lara, Maria do Carmo Custodio Souza Hunold; Cunha, Elenice Maria Siquetin; de Oliveira, Anderson Castro Soares; Braga, Ísis Assis; Aguiar, Daniel Moura

    2014-01-01

    The prevalence of antibodies against Equine Influenza Virus (EIV) was determined in 529 equines living on ranches in the municipality of Poconé, Pantanal area of Brazil, by means of the hemagglutination inhibition test, using subtype H3N8 as antigen. The distribution and possible association among positive animal and ranches were evaluated by the chi-square test, spatial autoregressive and multiple linear regression models. The prevalence of antibodies against EIV was estimated at 45.2% (95% CI 30.2 - 61.1%) with titers ranging from 20 to 1,280 HAU. Seropositive equines were found on 92.0% of the surveyed ranches. Equine from non-flooded ranches (66.5%) and negativity in equine infectious anemia virus (EIAV) (61.7%) were associated with antibodies against EIV. No spatial correlation was found among the ranches, but the ones located in non-flooded areas were associated with antibodies against EIV. A negative correlation was found between the prevalence of antibodies against EIV and the presence of EIAV positive animals on the ranches. The high prevalence of antibodies against EIV detected in this study suggests that the virus is circulating among the animals, and this statistical analysis indicates that the movement and aggregation of animals are factors associated to the transmission of the virus in the region. PMID:25351542

  19. Prevalence of antibodies to Toxoplasma gondii and Trichinella spiralis in 509 pigs from 31 farms in Oahu, Hawaii.

    PubMed

    Dubey, J P; Gamble, H R; Rodrigues, A O; Thulliez, P

    1992-06-01

    Serum samples from 509 pigs from 31 farms in Oahu, Hawaii were examined for antibodies to Toxoplasma gondii using the agglutination test in dilutions of 1:25, 1:50, and 1:500 and for Trichinella spiralis using the enzyme-linked immunosorbent assay. Toxoplasma gondii antibodies were found in 48.5% of pigs. Antibody titers were: 5.1% positive at 1:25 dilution, 28.6% positive at 1:50 dilution and 14.7% positive at 1:500 dilution. The prevalence of Toxoplasma gondii antibodies in garbage-fed pigs (67.3% of 199 pigs) was higher than in grain-fed pigs (33.8% of 180 pigs). Antibodies to Trichinella spiralis were found on seven of 31 farms. On five of these farms only a single serum was positive for Trichinella spiralis antibodies. The two remaining farms each had three positive sera (three of ten and three of 25 pigs tested); both of these farms fed garbage to pigs. PMID:1496803

  20. Relative seroprevalence of cysticercus antigens and antibodies and antibodies to Taenia ova in a population sample in south India suggests immunity against neurocysticercosis.

    PubMed

    Jayaraman, T; Prabhakaran, V; Babu, P; Raghava, M Venkata; Rajshekhar, V; Dorny, P; Muliyil, J; Oommen, A

    2011-03-01

    We evaluated the exposure of a community in Vellore district of south India to Taenia solium infection and its relationship to the prevalence of neurocysticercosis (NCC) causing active epilepsy. Seroprevalence of Taenia cysticercus antigens and antibodies were determined in 1064 randomly chosen asymptomatic individuals, antibodies to T. solium ova in 197 selected sera, and prevalence of taeniasis by a coproantigen test in 729 stool samples. The prevalence of NCC causing active epilepsy in Vellore district was determined in a population of 50 617. Coproantigens were detected in 0.8% (6 samples), Taenia cysticercus antigens in 4.5% (48 sera) and cysticercus IgG antibodies in 15.9% (169 sera) of the population. Cysticercus antibodies were directed against relatively low molecular weight cyst glycoprotein antigens in 14.9% (158 sera) of the population. IgG antibodies to Taenia ova were found in 81 (41.1%) of the selected samples. Prevalence of NCC causing active epilepsy was 1.3 per 1000 population. These results show high exposure of the population to the parasite and a relatively high prevalence of active infections (4.5% antigen positives) but a low prevalence of NCC causing active epilepsy (0.13%). These findings may indicate that the population is protected against developing neurocysticercosis. IgG antibodies directed against Taenia ova and low molecular weight cyst antigens may contribute to protection. PMID:21216417

  1. [Environmental and immunological survey of sensitization with Japanese cedar (Cryotomeria japonica) pollen in children. Part 2. The clinical significance of Japanese cedar (Cryptomeria japonica) pollen specific IgE antibody and IgG4 antibody].

    PubMed

    Takagi, M

    1992-11-01

    The clinical manifestations and Japanese cedar (sugi in Japanese) pollen specific antibodies were studied in 340 children who had not received specific hyposensitization. Specific IgE antibodies were measured by radioallergosorbent test (RAST) and IgG4 antibody by enzyme linked immunosorbent assay (ELISA). There were 52 children (15%) with RAST scores of over 2, and 44 children (13%) with ELISA of over 101 mu/ml. The specific IgG4 level was significantly higher in the RAST positive group than in the negative group. The incidence of pollinosis among children with RAST negative and IgG4 below 100 mu/ml was 13.3%, while combinations of RAST negative and higher ELISA, RAST positive and low ELISA, and RAST positive and higher ELISA were 50.0+-62.5%. There were 5 children with RAST scores of 4, and 6 children with ELISA of over 201 mu/ml. Symptoms of pollinosis manifested in all of them. Since the children with low IgE and high IgG4 antibodies had comparatively more symptoms, the IgG4 antibodies might be reaginic. It was concluded that measurement of sugi specific IgG4 antibodies as well as IgE antibodies could provide useful data on pollinosis in children. PMID:1492787

  2. Antibodies to a streptococcal antigen in gingival capillary and crevicular fluid washings, collected from man and non-human primates.

    PubMed

    Lehner, T; Harty, W J; Doubleday, B

    1987-01-01

    The local gingival antibody response was examined in actively-immunized rhesus monkeys and in human natural immunization to the cell-surface streptococcal antigen (SA I/II). A technique was developed to collect gingival capillary and crevicular fluid washings (GCCFW) from monkeys and human subjects with clinically normal gingiva. The thin crevicular epithelium and the adjacent capillary plexus are pierced with a probe and the resulting mixture of capillary blood, tissue fluid and crevicular fluid are collected by repeated washing of the gingival crevice. Antibodies measured by a solid-phase radioimmunoassay revealed that the IgG class of anti-SA I/II antibodies in GCCFW from the entire gingiva were positively correlated with the corresponding serum antibodies. Antibody levels from six defined dento-gingival units showed a progressive increase in the anti-SA I/II antibody level from the incisor to the premolar and molar units. A significant negative correlation was found between the antibody level of the total GCCFW and past caries experience. A similar negative correlation was found between antibodies from the GCCFW and the DMFS index of the molar teeth. The results are consistent with the hypothesis that there is an independent local immune response in the dento-gingival unit, which is superimposed on the common circulating serum antibodies and sensitized lymphocytes. PMID:3310975

  3. Associations between Mycobacterium avium subsp. paratuberculosis antibodies in bulk tank milk, season of sampling and protocols for managing infected cows

    PubMed Central

    2013-01-01

    Background The objective of this study was to identify associations between the concentration of Mycobacterium avium subsp. paratuberculosis (MAP) antibodies in bulk milk and potential risk factors in herd management and herd characteristics, explaining high MAP antibody titers in milk. An extensive questionnaire was administered to 292 organic and conventional dairy farms from New York, Wisconsin and Oregon. Bulk milk samples were taken from each farm for MAP enzyme-linked immunosorbent assay (ELISA). A general linear model was constructed with MAP ELISA value as the outcome variable and the management factors and herd characteristics as independent variables, while at the same time controlling for the study design variables of state, herd size, and production system (organic or conventional). High bulk tank MAP ELISA value may be due to either a high prevalence of MAP in a herd with many cows contributing to the antibody titer or due to a few infected cows that produce large quantities of antibodies. Results Results of the regression models indicated that bulk milk ELISA value was associated with season of sampling and the presence or absence of protocols for managing MAP-positive cows. The concentration of MAP antibodies in bulk milk varied seasonally with a peak in the summer and low concentrations in the winter months. When compared to farms that had never observed clinical Johne’s disease, keeping MAP-positive cows or only culling them after a period of delay was associated with an increase in optical density. Conclusions The seasonal variation in MAP antibody titers, with a peak in the summer, may be due to a seasonal increase in MAP-bacterial load. Additionally, seasonal calving practices may contribute to seasonal fluctuations in MAP antibody titers in bulk tank milk. Keeping MAP-positive cows increases the antibody titer in bulk milk, likely due to direct antibody production in the infected cow and indirect triggering of antibody production in herdmates. PMID:24283287

  4. Bands, interbands and puffs in native Drosophila polytene chromosomes are recognized by a monoclonal antibody to an epitope in the carboxy-terminal tail of histone H1

    Microsoft Academic Search

    Ronald J. Hill; Fujiko Watt; Catherine M. Wilson; Theodora Fifis; P. Anne Underwood; Gordon Tribbick; H. Mario Geysen; Jean O. Thomas

    1989-01-01

    A monoclonal antibody was raised against Drosophila melanogaster histone H1. Immunoscreening of proteolytic cleavage fragments of H1 and of a set of all possible overlapping synthetic octapeptides corresponding to the amino acid sequence of H1, revealed that the antibody recognizes an epitope within the sequence 207VTAAKPKA214 near the centre of the carboxy-terminal tail. This antibody gives positive immunofluorescence over the

  5. Avian influenza in ovo vaccination with replication defective recombinant adenovirus in chickens: vaccine potency, antibody persistence, and maternal antibody transfer.

    PubMed

    Mesonero, Alexander; Suarez, David L; van Santen, Edzard; Tang, De-Chu C; Toro, Haroldo

    2011-06-01

    Protective immunity against avian influenza (AI) can be elicited in chickens in a single-dose regimen by in ovo vaccination with a replication-competent adenovirus (RCA)-free human adenovirus serotype 5 (Ad)-vector encoding the AI virus (AIV) hemagglutinin (HA). We evaluated vaccine potency, antibody persistence, transfer of maternal antibodies (MtAb), and interference between MtAb and active in ovo or mucosal immunization with RCA-free recombinant Ad expressing a codon-optimized AIV H5 HA gene from A/turkey/WI/68 (AdTW68.H5(ck)). Vaccine coverage and intrapotency test repeatability were based on anti-H5 hemagglutination inhibition (HI) antibody levels detected in in ovo vaccinated chickens. Even though egg inoculation of each replicate was performed by individuals with varying expertise and with different vaccine batches, the average vaccine coverage of three replicates was 85%. The intrapotency test repeatability, which considers both positive as well as negative values, varied between 0.69 and 0.71, indicating effective vaccination. Highly pathogenic (HP) AIV challenge of chicken groups vaccinated with increasing vaccine doses showed 90% protection in chickens receiving > or = 10(8) ifu (infectious units)/bird. The protective dose 50% (PD50) was determined to be 10(6.5) ifu. Even vaccinated chickens that did not develop detectable antibody levels were effectively protected against HP AIV challenge. This result is consistent with previous findings ofAd-vector eliciting T lymphocyte responses. Higher vaccine doses significantly reduced viral shedding as determined by AIV RNA concentration in oropharyngeal swabs. Assessment of antibody persistence showed that antibody levels of in ovo immunized chickens continued to increase until 12 wk and started to decline after 18 wk of age. Intramuscular (IM) booster vaccination with the same vaccine at 16 wk of age significantly increased the antibody responses in breeder hens, and these responses were maintained at high levels throughout the experimental period (34 wk of age). AdTW68.H5(ch)-immunized breeder hens effectively transferred MtAb to progeny chickens. The level of MtAb in the progenies was consistent with the levels detected in the breeders, i.e., intramuscularly boosted breeders transferred higher concentrations of antibodies to the offspring. Maternal antibodies declined with time in the progenies and achieved marginal levels by 34 days of age. Chickens with high maternal antibody levels that were vaccinated either in ovo or via mucosal routes (ocular or spray) did not seroconvert. In contrast, chickens without MtAb successfully developed specific antibody levels after either in ovo or mucosal vaccination. These results indicate that high levels of MtAb interfered with active Ad-vectored vaccination. PMID:21793447

  6. Naturally acquired anthrax antibodies in a cheetah (Acinonyx jubatus) in Botswana.

    PubMed

    Good, Kyle M; Houser, Annmarie; Arntzen, Lorraine; Turnbull, Peter C B

    2008-07-01

    An outbreak of anthrax in the Jwana Game Reserve in Jwaneng, Botswana, was first observed when three cheetahs (Acinonyx jubatus) died of the disease in November 2004. In the aftermath of this event, banked serum samples collected from 23 wild-caught cheetahs were examined, by the inhibition enzyme-linked immunoassay (ELISA), for antibodies to the protective antigen (PA) of Bacillus anthracis. Of the 23 cheetahs, 16 regularly accessed the reserve. Antibodies to PA were detected in one cheetah collected in May 2004, indicating the disease was occurring well before it was first noticed. This appears to be the first demonstration of naturally acquired anthrax antibodies in cheetahs. The finding of one antibody-positive animal amongst at least 16 potentially exposed individuals is consistent with existing reports that it is uncommon for cheetahs to develop natural immunity to anthrax. PMID:18689661

  7. Diagnosis of intestinal amebiasis using salivary IgA antibody detection.

    PubMed

    del Muro, R; Acosta, E; Merino, E; Glender, W; Ortiz-Ortiz, L

    1990-12-01

    This investigation sought to determine whether detection of salivary IgA antibodies to Entamoeba histolytica could identify intestinal amebic infections among 223 school children. Four groups of children were identified through coproparasitoscopic examination: E. histolytica as other parasites only (20%); and parasite-free (25%). The diagnostic accuracy of salivary IgA antibodies to an E. histolytica membrane extract was 91.5% (sensitivity, 85%; specificity, 98%), maintaining high predictive value at different prevalences. Also, a positive correlation (r = .753, P less than .001) was observed between fecal E. histolytica membrane antigen levels and salivary IgA antibody activity. Measurement of IgA antibodies in saliva may be useful in diagnosing intestinal infections with E. histolytica within a wide range of prevalences. Moreover, sampling of saliva may be a useful non invasive test for immunoepidemiologic surveys. PMID:2230266

  8. [Antibodies to alpha-2A interferon in children with juvenile respiratory papillomatosis].

    PubMed

    Kol'tsov, V D; Onufrieva, E K; Chireshkin, D G; Malinovskaia, V V; Ershov, F I

    1995-01-01

    Follow-up of 36 JRP children and 12 controls (as shown by solid-phase enzyme immunoassay) has revealed that antibodies to IFN-alpha 2a with titers 1:20 to 1:1280 were present in 73.7% (14 of 19) of patients after interferon therapy and in 5.8% (1 of 17) of those who have not received interferon. None of the controls had the antibodies. Among the patients who have received only recombinant IFN-alpha 2a 88.8% carried the antibodies. In leukocytic interferon-treated group this number made up 20%. The primary results evidence that there is no negative effect of INF antibodies on the treatment results in the doses and schemes used. Positive results of INF in JRP were not reported either. PMID:7502448

  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. A survey of hantavirus antibody in small-mammal populations in selected United States National Parks.

    PubMed

    Mills, J N; Johnson, J M; Ksiazek, T G; Ellis, B A; Rollin, P E; Yates, T L; Mann, M O; Johnson, M R; Campbell, M L; Miyashiro, J; Patrick, M; Zyzak, M; Lavender, D; Novak, M G; Schmidt, K; Peters, C J; Childs, J E

    1998-04-01

    Hantavirus activity in 39 National Parks in the eastern and central United States was surveyed by testing 1,815 small mammals of 38 species for antibody reactive to Sin Nombre virus. Antibody-positive rodents were found throughout the area sampled, and in most biotic communities. Antibody was detected in 7% of 647 deer mice (Peromyscus maniculatus), 2% of 590 white-footed mice (P. leucopus), 17% of 12 rice rats (Oryzomys palustris), 3% of 31 cotton rats (Sigmodon hispidus), and 33% of 18 western harvest mice (Reithrodontomys megalotis). Antibody was also found in three of six species of voles, and in one of 33 chipmunks (Tamias minimus). Prevalence among Peromyscus was highest in the northeast. Although few cases of hantavirus pulmonary syndrome have been identified from the eastern and central regions, widespread infection in reservoir populations indicates that potential exists for human infection throughout much of the United States. PMID:9574803

  11. Bispecific antibodies that mediate killing of cells infected with human immunodeficiency virus of any strain.

    PubMed Central

    Berg, J; Lötscher, E; Steimer, K S; Capon, D J; Baenziger, J; Jäck, H M; Wabl, M

    1991-01-01

    Although AIDS patients lose human immunodeficiency virus (HIV)-specific cytotoxic T cells, their remaining CD8-positive T lymphocytes maintain cytotoxic function. To exploit this fact we have constructed bispecific antibodies that direct cytotoxic T lymphocytes of any specificity to cells that express gp120 of HIV. These bispecific antibodies comprise one heavy/light chain pair from an antibody to CD3, linked to a heavy chain whose variable region has been replaced with sequences from CD4 plus a second light chain. CD3 is part of the antigen receptor on T cells and is responsible for signal transduction. In the presence of these bispecific antibodies, T cells of irrelevant specificity effectively lyse HIV-infected cells in vitro. Images PMID:1905015

  12. Bead Arrays for Antibody and Complement Profiling Reveal Joint Contribution of Antibody Isotypes to C3 Deposition

    PubMed Central

    Ayoglu, Burcu; Szarka, Eszter; Huber, Krisztina; Orosz, Anita; Babos, Fruzsina; Magyar, Anna; Hudecz, Ferenc; Rojkovich, Bernadette; Gáti, Tamás; Nagy, György; Schwenk, Jochen M.; Sármay, Gabriella; Prechl, József

    2014-01-01

    The development of antigen arrays has provided researchers with great tools to identify reactivities against self or foreign antigens from body fluids. Yet, these approaches mostly do not address antibody isotypes and their effector functions even though these are key points for a more detailed understanding of disease processes. Here, we present a bead array-based assay for a multiplexed determination of antigen-specific antibody levels in parallel with their properties for complement activation. We measured the deposition of C3 fragments from serum samples to reflect the degree of complement activation via all three complement activation pathways. We utilized the assay on a bead array containing native and citrullinated peptide antigens to investigate the levels of IgG, IgM and IgA autoantibodies along with their complement activating properties in serum samples of 41 rheumatoid arthritis patients and 40 controls. Our analysis revealed significantly higher IgG reactivity against the citrullinated fibrinogen ? and filaggrin peptides as well as an IgA reactivity that was exclusive for citrullinated fibrinogen ? peptide and C3 deposition in rheumatoid arthritis patients. In addition, we characterized the humoral immune response against the viral EBNA-1 antigen to demonstrate the applicability of this assay beyond autoimmune conditions. We observed that particular buffer compositions were demanded for separate measurement of antibody reactivity and complement activation, as detection of antigen-antibody complexes appeared to be masked due to C3 deposition. We also found that rheumatoid factors of IgM isotype altered C3 deposition and introduced false-positive reactivities against EBNA-1 antigen. In conclusion, the presented bead-based assay setup can be utilized to profile antibody reactivities and immune-complex induced complement activation in a high-throughput manner and could facilitate the understanding and diagnosis of several diseases where complement activation plays role in the pathomechanism. PMID:24797804

  13. Prevalence of vesicular exanthema of swine antibodies among feral mammals associated with the southern California coastal zones.

    PubMed

    Smith, A W; Latham, A B

    1978-02-01

    Serum-neutralizing antibodies to both vesicular exanthema of swine virus (VESV) and San Miguel sea lion virus (SMSV) were found in a number of animal species having an association with the southern California coastal zones. California sea lions (Zalophus californianus) had antibodies to 9 VESV types (A48, C52, D53, E54, F54, G55, I55, J56, and K56). Fur seals (Callorhinus ursinus) and elephant seal pups (Mirounga angustirostris) were tested for antibodies to 6 VESV types and all were negative. California gray whales (Eschrichtius robustus) were tested for antibodies to 9 VESV types, and sperm whale (Physeter catodon), finback whale (Balaenoptera physalus), and sei whale (Balaenoptera borealis) were tested for 6 VESV types. Among the last 4 species, antibodies were present for each VESV type except C52, and all species were positive for antibodies to 2 or more VESV types. Feral swine from both Santa Cruz Island and Santa Catalina Island were tested and antibodies were present for 8 of 9 VESV types and all SMSV types except SMSV-4. One donkey from San Miguel Island was positive for VESV I55 and 2 were positive for SMSV-2. PMID:629463

  14. Development of Rapid Immunochromatographic Test with Recombinant NcSAG1 for Detection of Antibodies to Neospora caninum in Cattle

    PubMed Central

    Liao, Min; Zhang, Shoufa; Xuan, Xuenan; Zhang, Guohong; Huang, Xiaohong; Igarashi, Ikuo; Fujisaki, Kozo

    2005-01-01

    An immunochromatographic test (ICT) with recombinant surface antigen 1 of Neospora caninum (NcSAG1) was developed for the rapid detection of antibodies to N. caninum in cattle. The ICT was used to clearly discriminate between immunofluorescent-antibody test (IFAT)-positive bovine sera and IFAT-negative bovine sera. Serum samples collected from cattle in Yanbian, China, were examined by the ICT. Of the 96 serum samples, 23 (24.0%) were positive by the ICT, and 19 (19.8%) samples were positive by a previously developed enzyme-linked immunosorbent assay (ELISA). Eighteen of 19 ELISA-positive samples were positive according to the ICT. A good agreement was found between the results of the ICT and the ELISA. The results presented here suggest that the ICT with recombinant truncated NcSAG1 fused to glutathione S-transferase is a useful and reliable method for the detection of antibodies to N. caninum in cattle. PMID:16002641

  15. Development of rapid immunochromatographic test with recombinant NcSAG1 for detection of antibodies to Neospora caninum in cattle.

    PubMed

    Liao, Min; Zhang, Shoufa; Xuan, Xuenan; Zhang, Guohong; Huang, Xiaohong; Igarashi, Ikuo; Fujisaki, Kozo

    2005-07-01

    An immunochromatographic test (ICT) with recombinant surface antigen 1 of Neospora caninum (NcSAG1) was developed for the rapid detection of antibodies to N. caninum in cattle. The ICT was used to clearly discriminate between immunofluorescent-antibody test (IFAT)-positive bovine sera and IFAT-negative bovine sera. Serum samples collected from cattle in Yanbian, China, were examined by the ICT. Of the 96 serum samples, 23 (24.0%) were positive by the ICT, and 19 (19.8%) samples were positive by a previously developed enzyme-linked immunosorbent assay (ELISA). Eighteen of 19 ELISA-positive samples were positive according to the ICT. A good agreement was found between the results of the ICT and the ELISA. The results presented here suggest that the ICT with recombinant truncated NcSAG1 fused to glutathione S-transferase is a useful and reliable method for the detection of antibodies to N. caninum in cattle. PMID:16002641

  16. Isolated IgA Anti-?2 Glycoprotein I Antibodies in Patients with Clinical Criteria for Antiphospholipid Syndrome

    PubMed Central

    Ruiz-García, Raquel; Serrano, Manuel; Ángel Martínez-Flores, José; Mora, Sergio; Morillas, Luis; Martín-Mola, María Ángeles; Morales, José M.; Paz-Artal, Estela; Serrano, Antonio

    2014-01-01

    Seronegative antiphospholipid syndrome (SNAPS) is an autoimmune disease present in patients with clinical manifestations highly suggestive of Antiphospholipid Syndrome (APS) but with persistently negative consensus antiphospholipid antibodies (a-PL). IgA anti-?2 Glycoprotein I (aB2-GPI) antibodies are associated with APS. However, they are not currently considered to be laboratory criteria due to the heterogeneity of published works and the use of poor standardized diagnostic systems. We have aimed to assess aPL antibodies in a group of patients with clinical manifestations of APS (C-APS) to evaluate the importance of the presence of IgA aB2GPI antibodies in APS and its relation with other aPL antibodies. Only 14% of patients with C-APS were positive for any consensus antibody, whereas the presence of isolated IgA aB2GPI antibodies was found in 22% of C-APS patients. In patients with arterial thrombosis IgA aB2GPI, antibodies were the only aPL antibodies present. Serologic profile in primary APS (PAPS) is different from systemic autoimmune disorders associated APS (SAD-APS). IgA aB2GPI antibodies are more prevalent in PAPS and IgG aB2GPI antibodies are predominant in SAD-APS. The analysis of IgA aB2GPI antibodies in patients with clinical manifestations of PAPS might avoid underdiagnosed patients and provide a better diagnosis in patients with SAD-APS. Laboratory consensus criteria might consider including analysis of IgA aB2GPI for APS diagnosis. PMID:24741618

  17. Residue-Level Prediction of HIV-1 Antibody Epitopes Based on Neutralization of Diverse Viral Strains

    PubMed Central

    Chuang, Gwo-Yu; Acharya, Priyamvada; Schmidt, Stephen D.; Yang, Yongping; Louder, Mark K.; Zhou, Tongqing; Kwon, Young Do; Pancera, Marie; Bailer, Robert T.; Doria-Rose, Nicole A.; Nussenzweig, Michel C.; Mascola, John R.; Kwong, Peter D.

    2013-01-01

    Delineation of antibody epitopes at the residue level is key to understanding antigen resistance mutations, designing epitope-specific probes for antibody isolation, and developing epitope-based vaccines. Ideally, epitope residues are determined in the context of the atomic-level structure of the antibody-antigen complex, though structure determination may in many cases be impractical. Here we describe an efficient computational method to predict antibody-specific HIV-1 envelope (Env) epitopes at the residue level, based on neutralization panels of diverse viral strains. The method primarily utilizes neutralization potency data over a set of diverse viral strains representing the antigen, and enhanced accuracy could be achieved by incorporating information from the unbound structure of the antigen. The method was evaluated on 19 HIV-1 Env antibodies with neutralization panels comprising 181 diverse viral strains and with available antibody-antigen complex structures. Prediction accuracy was shown to improve significantly over random selection, with an average of greater-than-8-fold enrichment of true positives at the 0.05 false-positive rate level. The method was used to prospectively predict epitope residues for two HIV-1 antibodies, 8ANC131 and 8ANC195, for which we experimentally validated the predictions. The method is inherently applicable to antigens that exhibit sequence diversity, and its accuracy was found to correlate inversely with sequence conservation of the epitope. Together the results show how knowledge inherent to a neutralization panel and unbound antigen structure can be utilized for residue-level prediction of antibody epitopes. PMID:23843642

  18. Identification and characterization of an anti-isoaspartic acid monoclonal antibody.

    PubMed

    Lehrman, S R; Hamlin, D M; Lund, M E; Walker, G A

    1992-12-01

    The deamidation and rearrangement of protein-bound asparagine residues occurs when peptides and proteins are exposed to acidic or alkaline aqueous media. Asn99 of bovine growth hormone (bGH) is readily modified via these mechanisms. We have generated a monoclonal antibody (MAb) that interacts with a bGH fragment that contains an isoaspartyl residue. To obtain this antibody, CAF1/J mice were immunized with [isoaspartyl99]-bGH(96-112) conjugated to BSA. Using a competitive ELISA assay, the interaction of this MAb to [isoaspartyl99]-bGH(96-112) has been observed to have an apparent Km of 150 nM. The corresponding native peptide and other bGH fragments do not bind to this antibody with high affinity. For example, the binding affinities of [Asp99]-bGH(96-112) and [Glu99]-bGH(96-112) to this antibody are 54- and 78-fold lower than the corresponding isoaspartyl peptide. The antibody also binds to bGH that is enriched in isoaspartic acid at position 99, but not to the unmodified protein. The binding epitope of the peptide has been further characterized by comparing the binding of bGH(96-112) analogues to the MAb. Alanine substitution at residues 99, 100, 101, and 103 reduce binding affinity to the antibody by more than 10(3)-fold. Replacement of valine with alanine at position 102 has much less impact on antibody affinity. Further experiments suggest that the relative insensitivity to this substitution is due to the structural similarity of these sidechains. Other isoaspartic acid-containing peptides not derived from the bGH sequence do not bind to the antibody. We conclude that the epitope binding site of this MAb is highly specific for 99-103 of [isoaspartyl99]-bGH (96-112).(ABSTRACT TRUNCATED AT 250 WORDS) PMID:1281635

  19. [Comparison of eight screening tests for ant-HCV antibody].

    PubMed

    Deguchi, Matsuo; Kagita, Masanori; Yamashita, Naoko; Nakano, Takasi; Tahara, Kazuko; Asari, Seishi; Iwatani, Yoshinori

    2002-09-01

    We compared eight HCV screening tests for detection of anti-HCV antibody; Ortho Quick Chaser HCV Ab (QC), Ortho HCV Ab ELISA III (ELISA), Ortho HVC Ab PA test III (PA), Lumipulse II Ortho HCV (LUMI), IMx HCV.DAINAPACKII (IMx), ARCHITECT HCV (ARCH), Immucheck.F-HCV C50 Ab (Immu), RANREAM HCV Ab Ex II (RAN). Sera from six hundred patients were examined by these eight screening tests. The positive rates of the eight screening tests were from 9.0% to 13.2%. Forty-five sera showed discrepant results between the eight screening tests, and about half of them showed weak positive reaction and/or false positive. Twenty-five of the forty-five sera were negative for ant-HCV antibody in the CHIRON RIBA III confirmatory test, and forty-four of them were negative for HCV-RNA in the PCR method. The agreement rates between the two reagents were from 95.5% to 99.2%, but were not always high between the two reagents that used similar antigen. The specificities and sensitivities evaluated by using the RIBA III confirmatory test were excellent in ELISA, LUMI, IMx, ARCH and Immu. Three BBI seroconversion panels were used to compare the positive readings in the initial stage of HCV infection by eight screening tests. ELISA and ARCH showed the earliest positive readings, and then IMx, LUMI = RAN, PA, QC and Immu in this order. These findings indicate that ELISA and ARCH were the most excellent in the sensitivity, specificity and early diagnosis of HCV infection. However, we must pay attention to the weak positive reaction in the screening tests, because there is a possibility of "false positive". PMID:12391674

  20. Antibodies to laminin in Chagas' disease

    PubMed Central

    1982-01-01

    We have found that sera from humans with Chagas' disease and Rhesus monkeys infected with Trypanosoma cruzi contain IgM and IgG antibodies, which react with structures in a variety of connective tissues. These antibodies react with laminin but not with various other purified connective tissue components like collagen types I, III, IV, and V, fibronectin, heparan sulfate (BM-1) proteoglycan, or chondronectin. The tissue-reacting antibodies were isolated by absorption to a laminin- Sepharose column. The bound fraction contained all the tissue-reacting antibodies. These antibodies strongly stained trypomastigotes and amastigotes, but weakly stained epimastigotes. These studies show that sera from T. cruzi-infected primates contain antilaminin antibodies, which may be produced by those host in response to a laminin-like molecule present in the parasite. PMID:6801186

  1. Detection of Human Anti-Flavivirus Antibodies with a West Nile Virus Recombinant Antigen Microsphere Immunoassay

    PubMed Central

    Wong, Susan J.; Demarest, Valerie L.; Boyle, Rebekah H.; Wang, Tian; Ledizet, Michel; Kar, Kalipada; Kramer, Laura D.; Fikrig, Erol; Koski, Raymond A.

    2004-01-01

    We report a new, suspended-microsphere diagnostic test to detect antibodies to West Nile (WN) virus in human serum and cerebrospinal fluid (CSF). The microsphere immunofluorescence assay can be performed in less than 3 h on specimens of ?30 ?l. A recombinant WN virus envelope (E) protein antigen is covalently coupled to fluorescent polystyrene microspheres. After incubation with diluted serum or CSF, antibodies bound to the E protein antigen are detected with fluorescently labeled anti-human immunoglobulin antibody and flow analysis in a dual-laser Luminex 100 instrument. Retrospective testing of 833 sera from New York patients with suspected viral encephalitis demonstrated concordance with results obtained with the traditional enzyme-linked immunosorbent assay for immunoglobulin G (IgG) antibodies to WN virus (kappa = 0.85). One hundred eighty-eight (22.4%) of the samples, which were collected from June to November 2002, tested positive for antibodies to WN virus in the microsphere assay. Specimens depleted of IgG with anti-IgG antibody were reassayed to measure anti-E protein IgM antibodies and to provide an indication of current or recent WN virus infection. The assay also detects antibodies to E proteins from related flaviviruses, including St. Louis encephalitis, Japanese encephalitis, and dengue viruses. The new microsphere immunoassay provides a sensitive and rapid alternative to traditional enzyme-linked immunosorbent assays that detect antibodies to flavivirus E proteins. This assay can aid physicians and public health workers in the management of outbreaks of WN virus and related flaviviruses. PMID:14715733

  2. Detection of human anti-flavivirus antibodies with a west nile virus recombinant antigen microsphere immunoassay.

    PubMed

    Wong, Susan J; Demarest, Valerie L; Boyle, Rebekah H; Wang, Tian; Ledizet, Michel; Kar, Kalipada; Kramer, Laura D; Fikrig, Erol; Koski, Raymond A

    2004-01-01

    We report a new, suspended-microsphere diagnostic test to detect antibodies to West Nile (WN) virus in human serum and cerebrospinal fluid (CSF). The microsphere immunofluorescence assay can be performed in less than 3 h on specimens of antibodies bound to the E protein antigen are detected with fluorescently labeled anti-human immunoglobulin antibody and flow analysis in a dual-laser Luminex 100 instrument. Retrospective testing of 833 sera from New York patients with suspected viral encephalitis demonstrated concordance with results obtained with the traditional enzyme-linked immunosorbent assay for immunoglobulin G (IgG) antibodies to WN virus (kappa = 0.85). One hundred eighty-eight (22.4%) of the samples, which were collected from June to November 2002, tested positive for antibodies to WN virus in the microsphere assay. Specimens depleted of IgG with anti-IgG antibody were reassayed to measure anti-E protein IgM antibodies and to provide an indication of current or recent WN virus infection. The assay also detects antibodies to E proteins from related flaviviruses, including St. Louis encephalitis, Japanese encephalitis, and dengue viruses. The new microsphere immunoassay provides a sensitive and rapid alternative to traditional enzyme-linked immunosorbent assays that detect antibodies to flavivirus E proteins. This assay can aid physicians and public health workers in the management of outbreaks of WN virus and related flaviviruses. PMID:14715733

  3. Gangliosides, Ab1 and Ab2 antibodies

    Microsoft Academic Search

    Alejandro López-Requena; Marco Bestagno; Cristina Mateo de Acosta; Michela Cesco-Gaspere; Ana María Vázquez; Rolando Pérez; Oscar R. Burrone

    2007-01-01

    P3 mAb is an IgM monoclonal antibody specific for N-glycolyl-containing gangliosides. The immunogenicity of the P3 idiotype has been previously described by immunizing syngeneic BALB\\/c mice with the purified murine IgM or the mouse-human chimeric IgG antibody. In the present work we study the antibody response against the idiotype of P3 mAb through immunization with DNA. We used small immune

  4. Reconciling the Structural Attributes of Avian Antibodies*

    PubMed Central

    Conroy, Paul J.; Law, Ruby H. P.; Gilgunn, Sarah; Hearty, Stephen; Caradoc-Davies, Tom T.; Lloyd, Gordon; O'Kennedy, Richard J.; Whisstock, James C.

    2014-01-01

    Antibodies are high value therapeutic, diagnostic, biotechnological, and research tools. Combinatorial approaches to antibody discovery have facilitated access to unique antibodies by surpassing the diversity limitations of the natural repertoire, exploitation of immune repertoires from multiple species, and tailoring selections to isolate antibodies with desirable biophysical attributes. The V-gene repertoire of the chicken does not utilize highly diverse sequence and structures, which is in stark contrast to the mechanism employed by humans, mice, and primates. Recent exploitation of the avian immune system has generated high quality, high affinity antibodies to a wide range of antigens for a number of therapeutic, diagnostic and biotechnological applications. Furthermore, extensive examination of the amino acid characteristics of the chicken repertoire has provided significant insight into mechanisms employed by the avian immune system. A paucity of avian antibody crystal structures has limited our understanding of the structural consequences of these uniquely chicken features. This paper presents the crystal structure of two chicken single chain fragment variable (scFv) antibodies generated from large libraries by phage display against important human antigen targets, which capture two unique CDRL1 canonical classes in the presence and absence of a non-canonical disulfide constrained CDRH3. These structures cast light on the unique structural features of chicken antibodies and contribute further to our collective understanding of the unique mechanisms of diversity and biochemical attributes that render the chicken repertoire of particular value for antibody generation. PMID:24737329

  5. Reconciling the structural attributes of avian antibodies.

    PubMed

    Conroy, Paul J; Law, Ruby H P; Gilgunn, Sarah; Hearty, Stephen; Caradoc-Davies, Tom T; Lloyd, Gordon; O'Kennedy, Richard J; Whisstock, James C

    2014-05-30

    Antibodies are high value therapeutic, diagnostic, biotechnological, and research tools. Combinatorial approaches to antibody discovery have facilitated access to unique antibodies by surpassing the diversity limitations of the natural repertoire, exploitation of immune repertoires from multiple species, and tailoring selections to isolate antibodies with desirable biophysical attributes. The V-gene repertoire of the chicken does not utilize highly diverse sequence and structures, which is in stark contrast to the mechanism employed by humans, mice, and primates. Recent exploitation of the avian immune system has generated high quality, high affinity antibodies to a wide range of antigens for a number of therapeutic, diagnostic and biotechnological applications. Furthermore, extensive examination of the amino acid characteristics of the chicken repertoire has provided significant insight into mechanisms employed by the avian immune system. A paucity of avian antibody crystal structures has limited our understanding of the structural consequences of these uniquely chicken features. This paper presents the crystal structure of two chicken single chain fragment variable (scFv) antibodies generated from large libraries by phage display against important human antigen targets, which capture two unique CDRL1 canonical classes in the presence and absence of a non-canonical disulfide constrained CDRH3. These structures cast light on the unique structural features of chicken antibodies and contribute further to our collective understanding of the unique mechanisms of diversity and biochemical attributes that render the chicken repertoire of particular value for antibody generation. PMID:24737329

  6. Specificity of secretory antibodies to bacterial immunogens.

    PubMed

    Ebersole, J L; Molinari, J A

    1976-01-01

    The present investigation examined the specificity of the salivary immune response of axenic and conventional mice to topically administered Salmonella typhi, S. gallinarum, and S. typhimurium. Specific antibacterial antibodies were determined by passive hemagglutination and bacterial agglutination. Reciprocal antibody titers up to 320 were detected in saliva from mice immunized and assayed with homologous antigens. Antibodies to heterologous immunogens exhibited lower mean titers of 10 to 20 under identical conditions. High concentrations of specific antibodies to the somatic (O) antigen were detected in the saliva of mice administered these microorganisms; however, no significant differences in serum antibody levels were detectable after oral immunization. Only low levels of specific antiflagellar (H) antibodies were demonstrated in the saliva of immunized mice, whereas mean reciprocal titers of 20 were observed in the serum. Antibodies to the Vi antigen of S. typhi were detected in the saliva and serum of only those mice administered formalin-treated S. typhi. Examination of the classes of antibody elicited by these organisms indicated that immunoglobulin A (IgA) was the predominant class in saliva against the O antigens. The salivary response to the H antigens was comprised of both IgG and IgA, whereas the specific serum immunoglobulins were consistent with a primary humoral immune reaction. Local antibodies formed in response to the Vi antigen were exclusively IgG. Serum immunoglobulins produced after peroral administration of the somatic and virulence antigens were limited to the IgM class. PMID:765284

  7. [Managing patients with therapeutic antibodies in odontostomatology].

    PubMed

    Demoersman, J; Soueidan, A; Corre, P; Pers, J O

    2014-06-01

    Immunotherapies, particularly therapeutic antibodies, are increasingly used in the treatment of many autoimmune or oncological diseases. Patients treated with therapeutic antibodies may present with an increased risk of infection or of osteonecrosis of the jaws (ONJ). There is currently no consensus on the management of patients treated with therapeutic antibodies. These treatments are mainly used in hospitals, but they have been increasingly prescribed in ambulatory treatment for patients undergoing oral care. It is therefore important to establish therapeutic precautions for these patients. We had for aim to describe these antibody therapies, their indications, their potentially adverse effects in the oral cavity and to review the latest recommendations. PMID:24797731

  8. Preparation of astatine-labeled monoclonal antibodies

    SciTech Connect

    Milesz, S.; Norseev, Yu.V.; Szucs, Z. [Joint Inst. for Nuclear Research, Dubna (Russian Federation)]|[Central Inst. of Physical Research, Budapest (Hungary)

    1995-07-01

    In the cationic state astatine forms a stable complex with diethylenetriaminepentaacetic acid. Thanks to this complex, astatine can be bound to monoclonal antibodies of the RYa{sub 1} type. The most favorable conditions for preparing astatine-labeled antibodies are established. The chromatographic analysis and electromigration experiments showed that astatine is firmly linked to a biomolecule in vitro and it did not escape from labeled monoclonal antibodies even under treatment with such highly effective astatine-complexing agent as thiourea. The immune activity of astatine-labeled antibodies did not change even after 20 h.

  9. Plant cardiac glycosides and digoxin Fab antibody.

    PubMed

    Cheung, K; Urech, R; Taylor, L; Duffy, P; Radford, D

    1991-10-01

    The potential application of the Digoxin Fab antibody (Wellcome Digibind) in the clinical management of plant poisoning was investigated. The cardiac glycoside contents of various Australian plants were studied using immunoassay techniques. The cross-reactions of the Fab antibody and two digoxin assay antibodies against extracts of these plants were also studied. Results obtained indicated that the Digibind antibody cross-reacted with a wide range of glycosides contained in Australian plants and therefore could be of use in the treatment of life-threatening plant poisoning. PMID:1931226

  10. Monoclonal Antibody Therapy for Cancer

    Microsoft Academic Search

    Christoph Rader

    \\u000a Since the approval of rituximab (Rituxan®) for the treatment of B-cell non-Hodgkin’s lymphoma (B-NHL) in 1997, nine additional monoclonal antibodies (mAbs) have been\\u000a approved by the FDA for cancer therapy. Currently, more than 1,300 clinical studies registered at ClinicalTrials.gov investigate\\u000a mAb therapy of cancer, including more than 150 phase III clinical trials. In concert with their clinical acceptance, mAbs\\u000a in

  11. Camel Single-domain Antibodies as Modular Building Units in Bispecific and Bivalent Antibody Constructs

    Microsoft Academic Search

    Katja Els Conrath; Mark Lauwereys; Lode Wyns; Serge Muyldermans

    2001-01-01

    Single-domain antibodies against various antigens are isolated from the unique heavy-chain antibodies of immunized camels and llamas. These minimal sized binders are very robust and bind the antigen with high affinity in a monomeric state. We evaluated the feasibil- ity to produce soluble, functional bispecific and bivalent antibodies in Escherichia coli with camel single-domain antibody fragments as building blocks. Two

  12. Transplacental transfer of immune antibodies in the mouse demonstrated by antibody labeled in vivo with tritium

    E-print Network

    McKinney, Hubert Eugene

    1971-01-01

    TRANSPLACENTAL TRANSFER OF IMMUNE ANTIBODIES IN THE MOUSE DEMONSTRATED BY ANTIBODY LABELED IN VIVO WITH TRITIUM A Thesis by HUBERT EUGENE MCKINNEY Submitted to the Graduate College of Texas ASM University in partial fulfillment... of the requirement for the degree of MASTER OF SCIENCE August 1971 Major Sub] ect: Laboratory Animal Medicine TRAYSPLACENTAL TRANSFER OF IMMUNE ANTIBODIES IN THE MOUSE DEMONSTRATED BY ANTIBODY LABELED IN VIVO WITH TRITIUM A Thesis by HUBERT EUGENE MCKINNEY...

  13. Engineered antibody fragments and the rise of single domains

    Microsoft Academic Search

    Philipp Holliger; Peter J Hudson

    2005-01-01

    With 18 monoclonal antibody (mAb) products currently on the market and more than 100 in clinical trials, it is clear that engineered antibodies have come of age as biopharmaceuticals. In fact, by 2008, engineered antibodies are predicted to account for >30% of all revenues in the biotechnology market. Smaller recombinant antibody fragments (for example, classic monovalent antibody fragments (Fab, scFv))

  14. Position Announcement Fellowship/Contract Position

    E-print Network

    Yamamoto, Keith

    Position Announcement Fellowship/Contract Position Epidemiologist ­ Monitoring & Evaluation. This 2-year contract position will be located in the Epidemiology and Strategic Information Branch (ESIB Conduct a review of existing tools, draft minimum data elements, draft data collection tools, recommend

  15. Prevalence of antibodies to Legionella pneumophila in animal populations.

    PubMed Central

    Collins, M T; Cho, S N; Reif, J S

    1982-01-01

    We examined more than 2,800 human and animal sera for antibodies to four serogroups of Legionella pneumophila by using the microagglutination test. Antibody titers of greater than or equal to 1:64 were considered positive. The occurrence of positive equine sera (31.4%) was significantly higher than the occurrence of positive sera in cattle (5.1%), swine (2.9%), sheep (1.9%), dogs (1.9%), goats (0.5%), wildlife (0%), and humans (0.4%). The highest titer measured in horses was 1:512. The occurrence of positive sera in horses was related directly to age. In horses less than or equal to 1, 2 to 3, 4 to 7, 8 to 12, and greater than or equal to 13 years old, the percentages of positive sera were 0, 10.1, 30.3, 44.9 and 58.1%, respectively. When we compared age-specific serogroup-specific rates in horses from Colorado and Pennsylvania, we found differences. With horses 8 to 12 and greater than or equal to 13 years old, there was a significantly higher (P less than 0.05) occurrence of sera that reacted to serogroups II and III in horses from Pennsylvania. Of 242 positive sera, 43.8% reacted to a single serogroup (serogroup III or I most commonly), and 56.2% reacted to multiple serogroups (serogroups II and III or serogroups I, II, and III most commonly). A high percentage of seropositive horses suggested that horses are commonly infected with L. pneumophila or related organisms, and the age-specific rates of occurrence indicated that infection was related directly to duration of exposure. A definitive demonstration of equine infection will depend on isolation of the agent and repetition of this serological study with antigens obtained from organisms isolated from horses. PMID:7186901

  16. Monoclonal Antibodies Identify Novel Neural Antigens

    NASA Astrophysics Data System (ADS)

    Hawkes, Richard; Niday, Evelyn; Matus, Andrew

    1982-04-01

    Monoclonal antibodies (Mabs) were raised against synaptic plasma membranes from rat cerebellum. The hybridomas were screened with a solid-phase immunoassay, the positive lines were characterized by their immunoperoxidase staining pattern on cerebellum, and the specific polypeptide antigens were identified on protein blots. Among the Mabs described are some that stain only neurons or only glia and others that react with specific parts of cells, such as axons, dendrites, and synapses. Many Mabs reveal novel relationships between antigens and the cells in which they occur. For example, a Mab designated 7D5 reacts with a family of > 30 proteins but stains only glial cells. Several Mabs stain punctate sites of synaptic size and distribution in the cerebellar cortex but each reacts with a different subset of polypeptides. One of the most restricted cytological staining patterns is given by 12D5, which stains punctate sites in the granular layer of the cerebellar cortex and reacts with a single polypeptide band of apparent Mr 270,000. These results illustrate the feasibility of raising Mabs that can be used to follow the expression of specific gene products during brain development.

  17. Positioning Therapy for Ulcerative Colitis.

    PubMed

    Grinspan, Ari; Kornbluth, Asher

    2015-08-01

    Ulcerative colitis (UC) is a chronic inflammatory condition of the colon, characterized by diffuse mucosal inflammation, bloody diarrhea, and urgency. The mainstay of treatment has been mesalamine agents, steroids, thiopurines, and anti-tumor necrosis factor alpha (TNF-?) antibodies. Over the past several years, new therapies have emerged which have provided clinicians new treatment options as well as new challenges in deciding which treatment is best for their patient at given points in their disease course. These agents include budesonide-Multi-Matrix System (MMX), adalimumab, golimumab, and vedolizumab. In addition, randomized controlled trials have investigated a combination therapy of infliximab and azathioprine and a controlled trial of infliximab versus cyclosporine for intravenous steroid refractory UC. This review will focus on where these agents may be optimally positioned in treatment algorithms for UC. PMID:26143627

  18. Structural aspects of human leukocyte antigen class I epitopes detected by human monoclonal antibodies.

    PubMed

    Duquesnoy, Rene J; Marrari, Marilyn; Mulder, Arend; Claas, Frans H J; Mostecki, Justin; Balazs, Ivan

    2012-03-01

    This study addresses the concept that human leukocyte antigen (HLA) class I-specific alloantibodies are specific for epitopes that correspond to HLAMatchmaker-defined eplets. Eplets are essential parts of so-called structural epitopes that make contact with the 6 complementarity determining regions of an antibody. From published molecular models of crystallized protein antigen-antibody complexes, we have calculated that contact residues on structural HLA epitopes should reside within a 15-Å radius of a mismatched eplet. This study addresses the structural basis of high-frequency HLA class I epitopes reacting with human monoclonal antibodies (mAbs) derived from women sensitized during pregnancy. All mAbs were tested in Luminex assays with single HLA allele panels. The HLAMatchmaker algorithm was used to determine their specificity in context with eplet sharing between the immunizing allele and antibody-reactive alleles. To assess the autoreactive B cell origin of these antibodies, we have applied the recently developed nonself-self paradigm of epitope immunogenicity to analyze residue differences between the immunizer and the alleles of the antibody producer. A total of 9 mAbs were specific for epitopes associated with the 41T, 80NRG, 163LW, 69AA, or 80ERILR eplets. In each case, the immunizing allele had within 15 Å of the mismatched eplet, no residue differences with 1 of the alleles of the antibody producer. This observation is consistent with the concept that these mAbs originated from B cells with self HLA immunoglobulin receptors. Eplet-carrying alleles exhibited different levels of reactivity, which, when compared with the immunizing allele, ranged from high to intermediate to very low. In many cases, lower reactivities were associated with differences from self to nonself residues in surface locations within 15 Å of the specific eplet. Apparently, such locations may serve as critical contact sites for the antibody. In other cases, other residue differences did not appear to affect binding with the antibody, suggesting that these locations do not play a major role in antibody binding. For these mAbs we did not obtain convincing evidence that residue differences in hidden positions below the molecular surface had significant effects on antibody binding. These findings have increased our understanding of the structural basis of the immunogenicity and antigenicity of HLA class I epitopes and provide a basis for interpreting HLA antibody reactivity patterns in Luminex assays with single alleles. PMID:22227099

  19. Thyroid hormone antibodies and Hashimoto's thyroiditis in mongrel dogs

    SciTech Connect

    Rajatanavin, R.; Fang, S.L.; Pino, S.; Laurberg, P.; Braverman, L.E.; Smith, M.; Bullock, L.P.

    1989-05-01

    Abnormally elevated serum T3 concentrations measured by RIA were observed in 19 clinically euthyroid or hypothyroid mongrel dogs. The serum T4 concentrations in these sera were low, normal, or high. Measurement of the intensity of thyroid hormone binding to serum proteins was determined by equilibrium dialysis. A marked decrease in the percent free T3 was observed in these abnormal sera. Polyacrylamide gel electrophoresis, pH 7.4, of normal dog serum enriched with tracer /sup 125/I-labeled thyroid hormones demonstrated binding of (/sup 125/I)T4 to transthyretin, thyroid hormone-binding globulin, and albumin and of (/sup 125/I)T3 primarily to thyroid hormone-binding globulin. In all abnormal sera, polyacrylamide gel electrophoresis demonstrated strikingly higher binding of T3 to immunoglobulin (Ig). Eleven of 16 abnormal sera had minimal to moderate binding of T4 to Ig. The percent free T4 was lower only in dogs whose sera demonstrated markedly increased binding of T4 to Ig. All abnormal sera tested had positive antithyroglobulin antibodies, consistent with the diagnosis of autoimmune lymphocytic thyroiditis. As in humans, antibodies to thyroid hormones in dogs are more common in the presence of Hashimoto's thyroiditis and should be considered when elevated serum thyroid hormone concentrations are observed in the absence of clinical thyrotoxicosis. When an antibody to only one thyroid hormone is present, a marked discrepancy in the serum concentrations of T3 and T4 will be observed.

  20. STAINING TOXOPLASMA GONDII WITH FLUORESCEIN-LABELLED ANTIBODY

    PubMed Central

    Goldman, Morris

    1957-01-01

    A new serologic test for antibodies to Toxoplasma is described, which is based upon inhibition of specific staining with fluorescent antibody. In performing the test, a mixture of the test serum and known fluorescein-labelled antiserum is added to a dried smear of toxoplasms for 1 hour at 37°C. The smear is then rinsed and examined with a fluorescence microscope. Reduction in the brightness of fluorescence, as compared to that of a negative control slide, indicates the presence of antibody in the test serum. A comparison of the results of this test with those of the methylene blue dye test showed a strong parallelism between the two sets of results. On the other hand, the complement-fixation test for toxoplasmosis did not yield nearly as many positives as the inhibition test. The specificity of the new test was studied by comparing it with dye test results and clinical histories in human patients, and by testing a group of animals immunized with a variety of non-Toxoplasma antigens. No evidence of cross-reactions was obtained in the latter series. Some advantages and disadvantages of the inhibition test are discussed. PMID:13428924

  1. Neutralising antibodies for Mayaro virus in Pantanal, Brazil

    PubMed Central

    Pauvolid-Corrêa, Alex; Juliano, Raquel Soares; Campos, Zilca; Velez, Jason; Nogueira, Rita Maria Ribeiro; Komar, Nicholas

    2015-01-01

    The Pantanal hosts diverse wildlife species and therefore is a hotspot for arbovirus studies in South America. A serosurvey for Mayaro virus (MAYV), eastern (EEEV), western (WEEV) and Venezuelan (VEEV) equine encephalitis viruses was conducted with 237 sheep, 87 free-ranging caimans and 748 equids, including 37 collected from a ranch where a neurologic disorder outbreak had been recently reported. Sera were tested for specific viral antibodies using plaque-reduction neutralisation test. From a total of 748 equids, of which 264 were immunised with vaccine composed of EEEV and WEEV and 484 had no history of immunisation, 10 (1.3%) were seropositive for MAYV and two (0.3%) for VEEV using criteria of a ? 4-fold antibody titre difference. Among the 484 equids without history of immunisation, 48 (9.9%) were seropositive for EEEV and four (0.8%) for WEEV using the same criteria. Among the sheep, five were sero- positive for equine encephalitis alphaviruses, with one (0.4%) for EEEV, one (0.4%) for WEEV and three (1.3%) for VEEV. Regarding free-ranging caimans, one (1.1%) and three (3.4%), respectively, had low titres for neutralising antibodies to VEEV and undetermined alphaviruses. The neurological disorder outbreak could not be linked to the alphaviruses tested. Our findings represent strong evidence that MAYV and all equine encephalitis alphaviruses circulated in the Pantanal. PMID:25742272

  2. Comparison of an indirect fluorescent antibody test with Western blot for the detection of serum antibodies against Encephalitozoon cuniculi in cats.

    PubMed

    Künzel, Frank; Peschke, Roman; Tichy, Alexander; Joachim, Anja

    2014-12-01

    Current clinical research indicates that Encephalitozoon (E.) cuniculi infections in cats may be underdiagnosed, especially in animals with typical ocular signs (cataract/anterior uveitis). Although molecular detection of the pathogen in tissue appears promising, serology remains the major diagnostic tool in the living animal. While serological tests are established for the main host of E. cuniculi, the rabbit, the routine serological diagnosis for cats still needs validation. The aim of the study was to evaluate the consistency of indirect fluorescence antibody test (IFAT) and Western blot (WB) for the detection of IgG antibodies against E. cuniculi in the serum of 84 cats. In addition, PCR of liquefied lens material or intraocular fluid was performed in those of the cats with a suspected ocular E. cuniculi infection. Twenty-one cats with positive PCR results were considered as a positive reference group. Results obtained by IFAT and WB corresponded in 83/84 serum samples, indicating a very good correlation between both serological methods. Using WB as the standard reference, sensitivity and specificity for the detection of antibodies against E. cuniculi by the IFAT were 97.6 and 100%, respectively. The positive and negative predictive values for the IFAT were 100 and 97.7%, respectively. The accuracy (correct classified proportion) for the detection of IgG antibodies against E. cuniculi in cats was 98.8%. The comparison of both serological methods with the PCR results also revealed a good agreement as 20 out of 21 PCR-positive samples were seropositive both in IFAT and WB. Both tests can be considered as equally reliable assays to detect IgG antibodies against E. cuniculi in cats. As the IFAT is quicker and easier to perform, it is recommended for routine use in the diagnosis of feline encephalitozoonosis. PMID:25199557

  3. Detection of anticentromere antibodies using cloned autoantigen CENP-B.

    PubMed

    Rothfield, N; Whitaker, D; Bordwell, B; Weiner, E; Senecal, J L; Earnshaw, W

    1987-12-01

    A solid-phase enzyme-linked immunosorbent assay has been established using a cloned fusion protein, CtermCENP-B [beta-gal], as antigen. The fusion protein carries the major epitope of CENP-B, the major centromeric autoantigen. The enzyme-linked immunosorbent assay was more sensitive than immunofluorescence techniques in detecting anticentromere antibodies in patients with scleroderma or Raynaud's disease, and was weakly positive in 3% of normal controls and in 3% of 70 patients with other connective tissue diseases. PMID:3435569

  4. Monoclonal antibodies to the cell surface and a soluble form of the human nerve growth factor receptor

    SciTech Connect

    Clagett-Dame, M.; Chung, C.; Chao, M.V.; DiStefano, P.S. (Univ. of Wisconsin, Madison (USA))

    1990-12-01

    Monoclonal antibodies (designated IIIG5, VIID1, VIIIC8, and XIF1) have been produced that bind to the human nerve growth factor receptor (NGF-R) as well as to a soluble, truncated form of the receptor (NGF-Rt). The antibodies were generated against partially purified NGF-Rt from the conditioned medium of E9b cells, a transfected mouse fibroblast cell line (Ltk-) that expresses large numbers of the low affinity form of the human NGF-R on its cell surface. Hybridomas were screened by radiometric immunosorbent assay (RISA) and by immunoprecipitation of solubilized cell surface receptor covalently cross-linked to {sup 125}I-NGF. Four positive lines were cloned by limiting dilution and were found to secrete monoclonal antibodies of the IgGl,k subclass. All monoclonal antibodies bound to both NGF-R and NGF-Rt. Two monoclonal antibodies (VIID1, XIF1) immunoblotted the NGF-R from E9b cell preparations resolved on non-reducing sodium dodecyl sulfate (SDS)-polyacrylamide gels. The antibodies immunoprecipitated NGF-R from both E9b cells and from SH-SY5Y human neuroblastoma cells. The monoclonal antibodies bound to monkey (rhesis and cynomolgus) NGF-Rt, but did not cross-react with NGF-R from chick or rat. Results of antibody competition studies demonstrated that three antibodies bound to a similar or overlapping epitope on the NGF-Rt and one monoclonal antibody (IIIG5) recognized a distinct receptor epitope. Antibodies that bound to different sites on the receptor were used to develop a sensitive 2-site RISA. The 2-site RISA can be used to rapidly quantitate NGF-R and NGF-Rt in large numbers of biological samples in the absence of added {sup 125}I-labeled NGF.

  5. Antibodies Act Jointly to Promote Inflammation in Rheumatoid Arthritis

    MedlinePLUS

    ... ease the symptoms. Molecules called antibodies usually help combat infections by binding to foreign molecules like viral ... the antibodies to bind to their normal human targets, creating what are known as complexes. These antibody- ...

  6. Radiohalogenated half-antibodies and maleimide intermediate therefor

    DOEpatents

    Kassis, A.I.; Khawli, L.A.

    1991-02-19

    N-(m-radiohalophenyl) maleimide can be conjugated with a reduced antibody having a mercapto group to provide a radiolabeled half-antibody having immunological specific binding characteristics of whole antibody. No Drawings

  7. Bioconjugation of antibodies to horseradish peroxidase (hrp)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The bioconjugation of an antibody to an enzymatic reporter such as horseradish peroxidase (HRP) affords an effective mechanism by which immunoassay detection of a target antigen can be achieved. The use of heterobifunctional cross—linkers to covalently link antibodies to HRP provides a simple and c...

  8. Complement in antibody-based tumor therapy.

    PubMed

    Derer, Stefanie; Beurskens, Frank J; Rosner, Thies; Peipp, Matthias; Valerius, Thomas

    2014-01-01

    Monoclonal antibodies constitute a major treatment option for many tumor patients. Due to their specific recognition sites in their constant Fc regions, antibodies are able to trigger antibody-dependent cell-mediated cytotoxicity (ADCC) or complement-dependent cytotoxicity (CDC). While the contribution of ADCC to clinical efficacy has been strengthened by observations that patients with favorable Fc? receptor polymorphisms display better response rates to therapeutic antibodies, the contribution of CDC to their clinical efficacy remains controversial. In the background of high expression of complement-regulatory proteins on tumor cells as well as of the fact that some therapeutic antibodies lack the capacity to trigger efficient CDC, strategies have been implemented to improve either the capacity of antibodies to initiate the complement cascade or to interfere with tumor cells' resistance mechanisms. Although both strategies have demonstrated therapeutic benefit in vitro and in murine models, CDC-enhanced antibodies-to the best of our knowledge-have not been clinically tested, and evidence for the potential of CDC-optimizing approaches has yet to be generated in humans. Hence, the potency of complement activation and its impact on the clinical efficacy of therapeutic antibodies still remains to be elucidated in clinical trials encompassing novel complement-enhancing molecules. PMID:24941073

  9. Induction and detection of antibodies to squalene

    Microsoft Academic Search

    Gary R Matyas; Mangala Rao; Carl R Alving

    2002-01-01

    An improved high throughput assay for measuring murine antibodies to squalene (SQE) is described. The assay is highly reproducible and sensitive and can detect 80 ng\\/ml of antibody to SQE. The assay, an ELISA, is similar to our previously described assay in which plates containing PVDF membranes were used [J. Immunol. Methods 245 (2000) 1]. The PVDF plates worked well

  10. Structure and specificity of lamprey monoclonal antibodies

    E-print Network

    Ronquist, Fredrik

    anthracis spores. The recombinant VLR-B antibodies possess 8­10 uniform subunits that collectively bind describe the production of recombinant VLR-B antibodies specific for BclA, a major coat protein of Bacillus abortus, and human red blood cells (6­10). More recently, we demonstrated that immunization with Bacillus

  11. Original article Production of monoclonal antibodies against

    E-print Network

    Paris-Sud XI, Université de

    Original article Production of monoclonal antibodies against equine influenza : application 1988) Summary ― Monoclonal antibodies (Mo Abs) were prepared against influenza/A/equine/Prague/1. These monoclonals were tested against the 2 reference strains, 8 field strains of equine influenza virus, 3 human

  12. How to successfully patent therapeutic antibodies.

    PubMed

    Lahrtz, Fritz

    2015-04-01

    Therapeutic antibodies have become an established class of drugs for the treatment of a variety of diseases, especially cancer and autoimmune/inflammatory disorders, and a sufficient patent protection is a prerequisite for their successful commercialization. As monoclonal antibodies and their therapeutic potential have been well known for decades, the mere production of yet another therapeutic antibody is in many jurisdictions not considered a patentable invention. In contrast, antibodies with novel structural features and/or improved properties may be patentable. When drafting the claims, care should be taken to obtain a broad patent scope that protects both the antibody of interest and related antibodies having the same functional features, thereby preventing competitors from marketing a functionally equivalent antibody. Furthermore, the application should contain experimental evidence showing the improved properties of the claimed antibody. After the filing of a priority patent application, patent protection should be initiated at least in countries that are of particular commercial importance. Subsequent inventions relating to novel uses, formulations, dosage regimens, and combinations with other treatment modalities should be protected by further patent applications to extend patent term. PMID:25614506

  13. Monoclonal antibodies specific for mercuric ions.

    PubMed Central

    Wylie, D E; Lu, D; Carlson, L D; Carlson, R; Babacan, K F; Schuster, S M; Wagner, F W

    1992-01-01

    Monoclonal antibodies (mAbs) that react with soluble mercuric ions have been produced by injection of BALB/c mice with a hapten-carrier complex designed to maximize exposure of the metal to the immune system. Three hybridomas producing antibodies that reacted with bovine serum albumin (BSA)-glutathione-HgCl, but not with BSA-glutathione, were isolated from the spleen of a mouse given multiple injections with glutathione-HgCl conjugated to keyhole limpet hemocyanin. Stable subclones were established from two of these antibodies, designated mAb 4A10 and mAb 1F10. The binding of both antibodies to immobilized BSA-glutathione-HgCl was inhibited by soluble HgCl2, and dissociation constants for mercuric chloride binding were 2.3 and 3.7 nM for mAbs 4A10 and 1F10, respectively. Both antibodies bound mercuric acetate with similar affinities, demonstrating that the antibodies were capable of binding to mercuric ions in the presence of a different counterion than the one used in the immunogen. Reactions were not observed with other metal cations by either antibody. These data demonstrate the successful induction of antibodies that react very specifically with mercuric ions in solution regardless of the presence of a carrier. PMID:1570337

  14. Cytolytic Antibodies to Melanocytes in Vitiligo

    Microsoft Academic Search

    Jian Cui; Yuko Arita; Jean-Claude Bystryn

    1993-01-01

    Patients with vitiligo have been found to have circulating antibodies to pigment cells. To evaluate the functional activity of these antibodies, a highly sensitive europium release assay was used to compare complement-mediated cytolysis of human melanocytes by sera of 56 patients with vitiligo (20 with active disease, 25 with inactive disease, 11 with unidentified disease activity) and 47 control individuals.

  15. DNA Repair, Antibody Diversity, and Aging

    Microsoft Academic Search

    R. C. Johnson; A. C. Wang

    1985-01-01

    Proposed relationships of DNA repair, mutation, and the process of generation of antibody diversity allow new insights into the mechanism of aging. Pathways of antibody development are reviewed with special attention to steps which generate diversity. The normal process of combinatorial fusion of V region gene segments (i.e. V, D, and J) coding for the entire V region, plus the

  16. Longitudinal Monitoring of the Development of Antifilarial Antibodies and Acquisition of Wuchereria bancrofti in a Highly Endemic Area of Haiti

    PubMed Central

    Hamlin, Katy L.; Moss, Delynn M.; Priest, Jeffrey W.; Roberts, Jacquelin; Kubofcik, Joseph; Gass, Katherine; Streit, Thomas G.; Nutman, Thomas B.; Eberhard, Mark L.; Lammie, Patrick J.

    2012-01-01

    Antifilarial antibody testing has been established as a sensitive and specific method of diagnosing lymphatic filariasis. However, the development of serological responses to specific filarial antigens and their relationship to acquisition of infection is poorly understood. In order to evaluate whether the development of antigen specific antifilarial antibodies precedes microfilaremia and antigenemia, we compared the antibody responses of serum samples collected between 1990 and 1999 from a cohort of 142 Haitian children followed longitudinally. Antigen status was determined using the Og4C3 ELISA and the presence of microfilaremia was detected using microscopy. Antibody responses to Wb123, a Wuchereria bancrofti L3 antigen, were measured using a Luciferase Immunoprecipitation System (LIPS) assay. Antibody responses to Bm14 and Bm33, Brugia malayi antigens and to a major surface protein (WSP) from Wolbachia were analyzed using a multiplex bead assay. Over follow-up, 80 (56%) of the children became antigen-positive and 30 (21%) developed microfilaremia. Detectable antibody responses to Bm14, Bm33, Wb123, and WSP developed in 95%, 100%, 92%, and 29% of children, respectively. With the exception of WSP, the development of antibody responses generally preceded detection of filarial antigen. Our results show that antifilarial antibody responses can serve as an important epidemiological indicator in a sentinel population of young children and thus, may be valuable as tool for surveillance in the context of lymphatic filariasis elimination programs. PMID:23236534

  17. THE DETECTION OF ROTAVIRUS SPECIFIC ANTIBODY IN COLOSTRUM AND MILK BY ELISA

    E-print Network

    Paris-Sud XI, Université de

    THE DETECTION OF ROTAVIRUS SPECIFIC ANTIBODY IN COLOSTRUM AND MILK BY ELISA D.J. ELLENS P.W. DE, The Netherlands Résumé DETECTION DES ANTICORPS SPECIFIQUES DU ROTAVIRUS DANS LE COLOSTRUM ET LE LAIT PAR LA spécifiques du rotavirus dans le colostrum. Les résultats obtenus étaient en corrélation positive avec ceux d

  18. ANTIBODY DYNAMICS IN HOLSTEIN FRIESIAN HEIFERS VACCINATED WITH BRUCELLA ABORTUS STRAIN 19, USING SEVEN SEROLOGICAL TESTS

    Microsoft Academic Search

    N. P. Aguirre; V. R. Vanzini; S. Torioni de Echaide; B. S. Valentini; G. De Lucca; C. Aufranc; A. Canal; A. Vigliocco; K. Nielsen

    2002-01-01

    The serological response induced by Brucella abortus strain 19 was evaluated in 52 Holstein females from a brucellosis-free herd using seven serological tests. Each calf was vaccinated at an age of 4 and 8 months old with 3 × 10 CFU B. abortus S19 and the antibody response was determined as the proportion of positive results to each test. The

  19. The Statistical Precision of Medical Screening Procedures: Application to Polygraph and AIDS Antibodies Test Data

    Microsoft Academic Search

    Joseph L. Gastwirth

    1987-01-01

    The increased use of screening tests for drug use or antibodies to the HTLV-III (AIDS) virus, as well as pre-employment polygraph testing, has raised concerns about the reliability of the results of these procedures. This paper reviews the mathematical model underlying the analysis of data from screening tests. In addition to the known formulas for the proportion of positive (negative)

  20. Cross-reaction of an anti-Cryptosporidium monoclonal antibody with sporocysts of Monocystis species.

    PubMed

    Bull, S; Chalmers, R; Sturdee, A P; Curry, A; Kennaugh, J

    1998-06-15

    The non-specific cross-reaction of a fluorescently labelled anti-Cryptosporidium monoclonal antibody was observed microscopically when testing faecal specimens from small mammals. The reactive particles were identified as sporocysts of the Gregarine family Monocystidae, and indicate that considerable care should be taken so that false positives are not recorded. PMID:9746290

  1. Anti–LAMP-2 Antibodies Are Not Prevalent in Patients With Antineutrophil Cytoplasmic Autoantibody Glomerulonephritis

    PubMed Central

    Brown, Michael C.; Smith, Rex Neal; Badhwar, Anshul K.; Parente, Oscar; Chung, Hyun chul; O’Dell, Donna; Bunch; McGregor, JulieAnne G.; Hogan, Susan L.; Hu, Yichun; Yang, Jia-Jin; Berg, Elisabeth A.; Niles, John; Jennette, J. Charles; Preston, Gloria A.; Falk, Ronald J.

    2012-01-01

    Lysosomal membrane protein 2 (LAMP-2) is a target of antineutrophil cytoplasmic autoantibodies (ANCA) in addition to the more commonly known targets proteinase 3 and myeloperoxidase. The prevalence of anti–LAMP-2 antibodies and their relationship to disease in ANCA glomerulonephritis are not well described. We measured anti–LAMP-2 reactivity in 680 sera samples (two academic centers) from patients with ANCA glomerulonephritis (n=329); those with ANCA-negative glomerulonephritis (n=104); those with fimbriated, gram-negative Escherichia coli urinary tract infection (n=104); disease controls (n=19); and healthy volunteers (n=124). With levels in healthy controls used to define a reference range, anti–LAMP-2 reactivity was present in 21% of ANCA sera from two of the centers; reactivity was present in 16% of the control group with urinary tract infection. Western blotting and immunofluorescence microscopy did not verify positivity. Titers of anti-myeloperoxidase and anti–proteinase 3 antibodies were 1500-fold and 10,000-fold higher than anti–LAMP-2 titers, respectively. There was no correlation between anti–LAMP-2 antibodies and disease activity. Furthermore, Wistar Kyoto rats injected with anti–LAMP-2 antibodies did not develop glomerulonephritis. In conclusion, antibodies that react with LAMP-2 may exist at very low titers in a minority of patients with ANCA disease. These data do not support a mechanistic relationship between anti–LAMP-2 antibodies and ANCA glomerulonephritis. PMID:22021709

  2. Agents of equine viral encephalomyelitis: correlation of serum and cerebrospinal fluid antibodies.

    PubMed

    Keane, D P; Little, P B; Wilkie, B N; Artsob, H; Thorsen, J

    1988-04-01

    A survey was conducted by testing 115 paired equine serum and cerebrospinal fluid samples by hemagglutination-inhibition for antibodies to Powassan and snowshoe hare viruses, and by virus neutralization for antibodies to equine herpesvirus type 1. Twenty-five samples were from horses with spontaneous neurological disease and the remainder from horses euthanized because of various nonneurological disorders. All sera and cerebrospinal fluids were negative for antibodies to Powassan virus. Fifty-one sera (44.3%) and 15 cerebrospinal fluids (13.0%) had antibodies to snowshoe hare virus. Ninety-eight sera (85.2%) and four cerebrospinal fluids (3.5%) were positive for antibodies to equine herpesvirus type 1. Powassan virus was inoculated intracerebrally into one, and intravenously into four ponies. Neurological signs associated with a nonsuppurative encephalomyelitis occurred in three ponies. Antibodies to Powassan virus were detected in sera of all animals but in cerebrospinal fluids of only two. Powassan virus was isolated from brain and spinal cord of only the intracerebrally inoculated animal. PMID:2836046

  3. Influenza virus-specific neutralizing IgM antibodies persist for a lifetime.

    PubMed

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

    2014-11-01

    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

  4. 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

  5. Survival of patient with late onset hepatic failure by living-related liver transplantation from maternal donor with incompatible blood type

    Microsoft Academic Search

    Akira Kojima; Hitoshi Takagi; Takehiko Abe; Seiji Sakurai; Naondo Sohara; Satoru Kakizaki; Takeaki Nagamine; Masatomo Mori; Masatoshi Matsunami; Toshihiko Ikegami; Yasuhiko Hashikura; Seiji Kawasaki; Masatoshi Makuuchi

    1998-01-01

    :   A 14-year-old girl with blood type B with late onset hepatic failure (LOHF) of unknown cause has survived through living-related\\u000a liver transplantation (LRLT). No hepatitis virus, including HAV, HBV, HCV, and HGV, was positive at the onset of LOHF. Autoimmune\\u000a hepatitis was thought to be the cause because of positive results for serum anti-nuclear antibody at 80 times dilution

  6. Comparison of the western blot assay with the neutralizing-antibody and enzyme-linked immunosorbent assays for measuring antibody to verocytotoxin 1.

    PubMed Central

    Reymond, D; Karmali, M A; Clarke, I; Winkler, M; Petric, M

    1997-01-01

    A Western blot (immunoblot) assay (WBA) was developed to detect immunoglobulin G (IgG) antibodies against Escherichia coli Verocytotoxin 1 (VT1) by using a chemiluminescence detection system. The assay was compared with a VT1-neutralizing-antibody (VT1-NAb) assay and an anti-VT1 IgG enzyme-linked immunosorbent assay (ELISA). When four human serum samples that were known to be positive by VT1-NAb assay and ELISA were titrated to the endpoint by the three assays, the WBA gave endpoint titers that were up to 8-fold higher than those by ELISA and up to 256-fold higher than those by the VT1-NAb assay. Of 32 serum samples that were known to be positive by VT1-NAb assay and ELISA, 31 (97%) were positive by WBA; the one sample with a discrepant result gave borderline results by the VT1-NAb assay and ELISA. Of 52 serum samples that were known to be negative by the VT1-NAb assay and ELISA, 50 (96%) were negative and 2 (4%) were positive by WBA. Of 44 serum samples that gave discrepant results by the VT1-NAb assay and ELISA, neither of the latter correlated with the results of WBA. In an investigation of 19 pairs of acute- and convalescent-phase serum samples from patients with hemolytic-uremic syndrome, 10 pairs that were positive by the VT1-NAb assay were also WBA positive, while 9 pairs that were NAb negative were also WBA negative. The WBA is inherently more specific and sensitive than either the NAb assay or the ELISA and may be used as a "gold standard" to detect IgG antibodies to VT1. Like the NAb assay and the ELISA for detecting antibodies to VT1, the WBA has little to offer in the diagnostic setting but is expected to play an important role in seroepidemiological studies. PMID:9041398

  7. Synthetic Antibodies for Reversible Cell Recognition

    NASA Astrophysics Data System (ADS)

    Zhou, Jing Zhou

    2011-12-01

    Antibody-mediated cell recognition plays a critical role in various biological and biomedical applications. However, strong antibody-cell interactions can lead to the difficulty of separating antibodies from the bound cells in a simple and non-destructive manner, which is often necessary to numerous applications such as cell sorting or separation. Thus, this thesis research is aimed to create an antibody-like nanomaterial with the function of reversible cell recognition It was hypothesized that nucleic acid aptamer and dendrimer could be used as fundamental structural components to develop an antibody-like nanomaterial. The aptamer functions as the binding site of an antibody; the dendrimer is used as a robust, defined nano-scaffold to support the aptamer and to carry small molecules (e.g., fluorophores). To test this hypothesis, a novel method was first developed to discover the essential nucleotides of full-length aptamers to mimic the binding sites of antibodies. The essential nucleotides were further conjugated with a dendrimer to synthesize a monovalent aptamer-dendrimer nanomaterial. The results clearly showed that the essential nucleotides could maintain high affinity and specificity after tethered on dendrimer surface. To further test the hypothesis that antibody-like nanomaterials can be rationally designed to acquire the capability of reversible cell recognition, an aptamer that was selected at 0 °C was used as a model to synthesize a "Y-shaped" nanomaterial by conjugating two aptamers to the same dendrimer. The results showed that the nanomaterial-cell interaction could be affected by the distance between two binding aptamers. In addition, the "Y-shaped" antibody-like nanomaterial could bind target cells more strongly than its monovalent control. Importantly, the strong cell-nanomaterial interaction could be rapidly reversed when the temperature was shifted from 0 °C to 37 °C. In summary, we developed a synthetic antibody that can not only mimic the functional structure and cell recognition capability of antibodies, but also possess specific features that natural antibodies do not possess. This study has opened a new avenue for developing synthetic antibodies and has also advanced the understanding of the functionality of multivalent nanomaterials. This novel synthetic antibody holds great potential for various biological and biomedical applications such as cell separation.

  8. The cancer recognition (CARE) antibody test.

    PubMed

    Thornthwaite, Jerry T; McDuffee, Emily C; Harris, Robert B; Secor McVoy, Julie R; Lane, I W

    2004-12-28

    The cancer recognition (CARE) antibody (Ab) test is a serologic assay for a specific IgM that is elevated in cancer patients. All tests are measured using an indirect enzyme-linked immunosorbent assay (ELISA) of human serum. The target polypeptide in the CARE Ab test is the IgM binding epitope (LT-11) of the CARE antigen (Ag) consisting of a 16 mer structure that has been produced synthetically. The mean relative concentration (MRC) is determined relative to standard, normalized human plasma. Non-parametric analysis showed median MRC values of healthy volunteers (HVs) with no history of cancer (n =47), family history of cancer (n = 126) and a previous cancer history (n = 24) to be 26, 34 and 46, respectively. It was determined that there was no significance found among the medians of the three HV groups (P = 0.53). The specificity of the HV types was between 87 and 98%. Benign/non-cancer surgical patients (n = 27) had a median value of 20 with a specificity of 96%. The cancer patients (n = 61) had a median value of 246 with a sensitivity of 89%. There was a significant difference between the HV and cancer patients (P < 0.0001) as well as between the benign/surgical non-cancerous group and cancer patients (P < 0.0001). The IgM antibody is heat stable at room temperature for two days versus being frozen at -80 degrees C (r2 = 0.97). Either serum or plasma samples may be used in the CARE Ab test (r2 = 0.92). The CARE Ab was almost exclusively IgM with no serum conversion to IgG in sequential measurements of patients with cancer over a six-month period. Preliminary data from patients undergoing post-operative cancer treatment showed that decreasing Ab levels revealed patients negative for residual cancer or undergoing remission, while relapsing patients show an increase in Ab levels. A return to a positive Ab level shortly after treatment is a poor prognostic sign while in advanced cancers the Ab levels may be depressed significantly. PMID:15533599

  9. Functional properties of nonhuman primate antibody to Porphyromonas gingivalis.

    PubMed Central

    Anderson, D M; Ebersole, J L; Novak, M J

    1995-01-01

    The nonhuman primate (NHP) serves as a useful model for examining the host-parasite interactions in Porphyromonas gingivalis-associated periodontal disease. This study determined the influence of NHP sera on (i) the direct killing of P. gingivalis, (ii) P. gingivalis-induced superoxide anion (O2-) release from human polymorphonuclear leukocytes (PMNs), and (iii) the ability of PMNs to bind and phagocytize P. gingivalis. Three types of NHP sera were utilized: (i) normal or baseline sera; (ii) sera obtained after ligature-induced periodontitis; and (iii) sera obtained following active immunization with formalinized P. gingivalis. All assays were performed with or without the addition of human complement. Significantly more (P < 0.01) direct killing of P. gingivalis occurred with immunized sera and complement than with any of the other treatments. The sera from ligature-induced periodontitis NHPs had significantly less (P < 0.03) killing capacity than the baseline sera, which contained natural antibody produced to P. gingivalis colonization. Sera from immunized NHPs were used to opsonize P. gingivalis and caused significantly greater (P < 0.01) levels of O2- release from PMNs. Finally, the sera from immunized NHPs significantly enhanced (P < 0.009) the uptake of P. gingivalis by PMNs, although binding of the bacteria to PMNs was similar among all three serum types. Active immunization of NHPs with P. gingivalis elicited a functional antibody that enhanced direct killing, positively influenced the activation of PMNs, and enhanced the ability of PMNs to phagocytize P. gingivalis. Moreover, antibody produced as a sequela of progressing periodontitis appeared to lack these functions. A wide variability in functional capacity of the sera from individual NHPs, which may contribute to an individual's susceptibility to P. gingivalis-induced disease, was noted. This variability suggested that results from functional tests of serum antibody may aid in predicting host susceptibility to disease and response to therapy. PMID:7642252

  10. Clearance of pathological antibodies using biomimetic nanoparticles

    PubMed Central

    Copp, Jonathan A.; Fang, Ronnie H.; Luk, Brian T.; Hu, Che-Ming J.; Gao, Weiwei; Zhang, Kang; Zhang, Liangfang

    2014-01-01

    Pathological antibodies have been demonstrated to play a key role in type II immune hypersensitivity reactions, resulting in the destruction of healthy tissues and leading to considerable morbidity for the patient. Unfortunately, current treatments present significant iatrogenic risk while still falling short for many patients in achieving clinical remission. In the present work, we explored the capability of target cell membrane-coated nanoparticles to abrogate the effect of pathological antibodies in an effort to minimize disease burden, without the need for drug-based immune suppression. Inspired by antibody-driven pathology, we used intact RBC membranes stabilized by biodegradable polymeric nanoparticle cores to serve as an alternative target for pathological antibodies in an antibody-induced anemia disease model. Through both in vitro and in vivo studies, we demonstrated efficacy of RBC membrane-cloaked nanoparticles to bind and neutralize anti-RBC polyclonal IgG effectively, and thus preserve circulating RBCs. PMID:25197051

  11. An affinity improved single-chain antibody from phage display of a library derived from monoclonal antibodies detects fumonisins by immunoassay.

    PubMed

    Hu, Zu-Quan; Li, He-Ping; Wu, Ping; Li, Ya-Bo; Zhou, Zhu-Qing; Zhang, Jing-Bo; Liu, Jin-Long; Liao, Yu-Cai

    2015-03-31

    Fumonisin B analogs, particularly FB1, FB2, and FB3, are major mycotoxins found in cereals. Single-chain fragment variable (scFv) antibodies represent a promising alternative immunoassay system. A phage-displayed antibody library derived from four monoclonal antibodies (mAbs) generated against FB1 was used to screen high binding affinity scFv antibodies; the best candidate was designated H2. Surface plasmon resonance measurements confirmed that the H2 scFv displayed a 82-fold higher binding affinity than its parent mAb. Direct competitive enzyme-linked immunosorbent assay demonstrated that the H2 antibody could competitively bind to free FB1, FB2, and FB3, with an IC50 of 0.11, 0.04, and 0.10 ?M, respectively; it had no cross-reactivity to deoxynivalenol, nivalenol and aflatoxin. Validation assays with naturally contaminated samples revealed a linear relationship between the H2 antibody-based assay results and chemical analysis results, that could be expressed as y=1.7072x+5.5606 (R(2)=0.8883). Homology modeling of H2 revealed a favorable binding structure highly complementary to the three fumonisins. Molecular docking analyses suggested that the preferential binding of the H2 scFv to FB2 was due to the presence of a hydrogen radical in its R1 position, leading to a proper electrostatic matching and hydrophobic interaction. The H2 scFv antibody can be used for the rapid, accurate, and specific detection of fumonisin contamination in agricultural samples. PMID:25813030

  12. Possible association between anti-Ro antibodies and myocarditis or cardiac conduction defects in adults with systemic lupus erythematosus

    Microsoft Academic Search

    D Logar; T Kveder; B Rozman; J Dobovisek

    1990-01-01

    In view of the association of congenital heart block with maternal antibody to cellular antigen Ro (SSA), and one report linking anti-Ro with myocarditis in a patient with myositis an association between anti-Ro antibodies and cardiac disease was sought in adults with systemic lupus erythematosus (SLE). Among 67 patients with SLE, of whom 36 were anti-Ro positive, a significantly higher

  13. Anti-CCP antibody test predicts the disease course during 3 years in early rheumatoid arthritis (the Swedish TIRA project)

    PubMed Central

    Kastbom, A; Strandberg, G; Lindroos, A; Skogh, T

    2004-01-01

    Objectives: To evaluate the diagnostic sensitivity of antibodies to cyclic citrullinated peptide (CCP) in recent onset rheumatoid arthritis (RA) at diagnosis and 3 years later, and to evaluate anti-CCP antibody as a predictor of the disease course during 3 years. Methods: 242 patients with recent onset (?1 year) RA were followed up regularly during 3 years after inclusion in the Swedish multicentre study "TIRA" 1996–98. Anti-CCP antibodies were analysed by an enzyme immunoassay (EIA). Rheumatoid factors (RFs) were analysed by latex agglutination and two isotype-specific (IgM and IgA) EIAs. Disease activity was assessed by plasma CRP, ESR, 28 joint disease activity score, and the physician's global assessment of disease activity. Functional ability was evaluated by the Health Assessment Questionnaire. Results: Overall, the diagnostic sensitivity of anti-CCP antibodies was 64% and the proportion of positive tests increased with the number of fulfilled classification criteria according to the American College of Rheumatology. The anti-CCP antibody results correlated with RF, but were better than RF as predictor of a more aggressive disease course. After 3 years 5/97 patients had changed anti-CCP status: 2 from negative to positive and 3 from positive to negative. The mean level of anti-CCP antibodies declined by 131 U/ml during the 3 year follow up (95% CI 34 to 228 U/ml). Conclusion: The anti-CCP antibody assay has a similar diagnostic sensitivity to that of RF in early RA, but is better as a predictor of the disease course over 3 years. Although the mean serum level declines, anti-CCP antibody positivity remains essentially unaltered 3 years after diagnosis and start of antirheumatic treatment. PMID:15308517

  14. Randomised controlled trial of aspirin and aspirin plus heparin in pregnant women with recurrent miscarriage associated with phospholipid antibodies (or antiphospholipid antibodies)

    PubMed Central

    Rai, R.; Cohen, H.; Dave, M.; Regan, L.

    1997-01-01

    OBJECTIVE: To determine whether treatment with low dose aspirin and heparin leads to a higher rate of live births than that achieved with low dose aspirin alone in women with a history of recurrent miscarriage associated with phospholipid antibodies (or antiphospholipid antibodies), lupus anticoagulant, and cardiolipin antibodies (or anticardiolipin antibodies). DESIGN: Randomised controlled trial. SETTING: Specialist clinic for recurrent miscarriages. SUBJECTS: 90 women (median age 33 (range 22-43)) with a history of recurrent miscarriage (median number 4 (range 3-15)) and persistently positive results for phospholipid antibodies. INTERVENTION: Either low dose aspirin (75 mg daily) or low dose aspirin and 5000 U of unfractionated heparin subcutaneously 12 hourly. All women started treatment with low dose aspirin when they had a positive urine pregnancy test. Women were randomly allocated an intervention when fetal heart activity was seen on ultrasonography. Treatment was stopped at the time of miscarriage or at 34 weeks' gestation. MAIN OUTCOME MEASURES: Rate of live births with the two treatments. RESULTS: There was no significant difference in the two groups in age or the number and gestation of previous miscarriages. The rate of live births with low dose aspirin and heparin was 71% (32/45 pregnancies) and 42% (19/45 pregnancies) with low dose aspirin alone (odds ratio 3.37 (95% confidence interval 1.40 to 8.10)). More than 90% of miscarriages occurred in the first trimester. There was no difference in outcome between the two treatments in pregnancies that advanced beyond 13 weeks' gestation. Twelve of the 51 successful pregnancies (24%) were delivered before 37 weeks' gestation. Women randomly allocated aspirin and heparin had a median decrease in lumbar spine bone density of 5.4% (range -8.6% to 1.7%). CONCLUSION: Treatment with aspirin and heparin leads to a significantly higher rate of live births in women with a history of recurrent miscarriage associated with phospholipid antibodies than that achieved with aspirin alone. PMID:9022487

  15. Seroprevalence of poliovirus antibodies among 7-month-old infants after 4 doses of oral polio vaccine in Sistan-va-Baluchestan, Islamic Republic of Iran.

    PubMed

    Izadi, S; Shahmahmoodi, S; Zahraei, S M; Dorostkar, F; Majdzadeh, S-R

    2015-02-01

    Despite high coverage rates of polio vaccine in the Islamic Republic of Iran, the seroconversion rates of infants may be inadequate. This study measured seroprevalence of antibodies against poliovirus serotypes 1 to 3 (PV1, PV2 and PV3) in 7-month-old infants who had received at least 4 doses of trivalent oral polio vaccine. A serosurvey was conducted in 2010 in rural areas of Chabahar, Sistan-va-Baluchestan province. Using cluster sampling, 72 eligible infants were tested for antibody against the 3 poliovirus serotypes according to WHO guidelines. Antibody titres ? 1:10 were considered positive. The seropositive rates for antibody against PV1, PV2 and PV3 were 84.7%, 95.8% and 70.8% respectively. Only 63.9% of participants were seropositive for antibodies against all 3 poliovirus serotypes. Except for PV2, the seroprevalence of antibody against the other 2 poliovirus serotypes, especially PV3, was unsatisfactory. PMID:25876819

  16. Iodine-131 labeled anti-CEA polyclonal antibody detection of gastrointestinal cancer

    SciTech Connect

    Nabi, H.A.; Hinkle, G.H.; Olsen, J.O.; Haagensen, D.A.; Thurston, M.O.; Mojzisik, C.; Houchens, D.; Martin, E.W. Jr.

    1984-01-01

    To localize gastrointestinal tumor, 31 patients were injected with 1.7-2.1 mCi I-131 anti-CEA baboon polyclonal antibody. Whole body imaging at 48, 72, and occasionally 96 hrs was performed with a Signa Camera (Technicare) peaked at 364 keV with 20% window. Additional spot views were usually obtained. No subtraction methods were used. All patients had surgical and pathological confirmation of the nuclear medicine studies. Labeled antibody images were positive in 15 (8 recurrent or metastatic colorectal, 2 gastric, 1 pancreatic, 1 primary colon, and 1 breast metastatic to chest wall). In 1, antibody images were positive for metastatic deposits in para-aortic lymph nodes, but negative for primary rectal tumor. True negative images were observed in 6; false negative images in 9 (4 liver metastases, 2 rectal, 1 pancreatic, 1 mesenteric lymph node metastasis, 1 bone metastasis). In all cases, no correlation existed between preoperative CEA serum levels and imaging. I-131 labeled anti-CEA polyclonal antibody imaging proved highly efficient in detecting gastric cancer (2/2) and moderately efficient in detecting recurrent colorectal cancer (8/15). On the other hand, the I-131 labeled polyclonal anti-CEA antibody imaging was of limited value in detecting colon cancer (1/9), pancreatic cancer (1/4) and metastatic liver disease (0/4).

  17. Antibodies to bluetongue viruses in animals imported into United States zoological gardens.

    PubMed Central

    House, J A; Groocock, C M; Campbell, C H

    1982-01-01

    Three hundred forty-five serum samples from 30 zoological animal species which had been imported into the United States were examined retrospectively for the presence of antibody to bluetongue viruses. Ninety eight (28.4%) were positive for antibody to bluetongue group antigen by the bluetongue agar gel immunodiffusion test. Bluetongue antibodies, most of which were against serotypes exotic to the United States, were detected in 13 animal species from Africa not previously reported to be infected by bluetongue virus. The lack of virus neutralizing antibody to any of the 20 known bluetongue virus types in four of the 28 positive serums studied may indicate the existence of new bluetongue virus serotypes, cross reactions with other orbiviruses or a more rapid decline of neutralizing than precipitating antibody. The possibility of recrudescence of bluetongue virus infection from some inapparently infected zoological animals and existence of a known bluetongue vector (Culicoides variipennis) in the United States would suggest that further assessment of bluetongue in zoological animals be made. PMID:6178486

  18. Hepatitis C antibody prevalence in blood donors in different governorates in Egypt.

    PubMed

    Arthur, R R; Hassan, N F; Abdallah, M Y; el-Sharkawy, M S; Saad, M D; Hackbart, B G; Imam, I Z

    1997-01-01

    Markers of hepatitis B virus (HBV) and hepatitis C virus (HCV) infections were sought in serum samples from 2644 blood donors in 24 of Egypt's 26 governorates. Of the 2644 samples, 656 (24.8%) were shown to contain anti-HCV immunoglobulin G antibody by Abbott second generation enzyme immunoassays (EIA). Of 85 EIA-positive samples tested by recombinant immunoblot assay, 72 (85%) were positive. HCV seroprevalence in the governorates ranged from zero to 38%; 15 governorates (62%) had an HCV antibody prevalence greater than 20%, and 6 (25%) greater than 30%. Governorates with higher sero-prevalences were located in the central and north-eastern Nile river delta, and south of Cairo in the Nile river valley. Subjects from areas in and adjoining the Sinai peninsula, in the eastern and western desert, and in southernmost Egypt, had the lowest prevalence of HCV antibody. The large urban governorates of Cairo and Alexandria had antibody prevalences of 19% and 11%, respectively. A total of 39.4% subjects had evidence of HBV infection (and-HBV core antigen total antibody). HCV infections were detected more frequently in donors with markers for HBV infections than in uninfected subjects (36% versus 18%, P < 0.001). PMID:9231192

  19. Antibody responses to natural rattlesnake envenomation and a rattlesnake toxoid vaccine in horses.

    PubMed

    Gilliam, Lyndi L; Carmichael, Robert C; Holbrook, Todd C; Taylor, Jennifer M; Ownby, Charlotte L; McFarlane, Dianne; Payton, Mark E

    2013-05-01

    Antivenom antibody titers following administration of rattlesnake venom for antivenom production in horses are well documented; however, antivenom antibody titers following natural rattlesnake envenomation in horses are not. Antibody titers produced in response to the commercially available rattlesnake venom vaccine are also not published. Our study objectives were to measure antivenom antibody titers in rattlesnake-bitten horses and compare them to titers in horses vaccinated with the rattlesnake venom vaccine. Additionally, titers were compared in pregnant versus nonpregnant horses to assess the affect of pregnancy on vaccine response and were measured pre- and postsuckle in foals of vaccinated mares to detect passive transfer of vaccine immunoglobulins. Blood samples were collected from 16 rattlesnake-bitten horses. Thirty-six horses (11 pregnant mares, 12 nonpregnant mares, 13 geldings) were vaccinated using a Crotalus atrox venom toxoid vaccine. Blood was collected before administering each vaccination and 30 days following the third vaccination. Blood was collected from foals of vaccinated mares pre- and postsuckle. All serum was assayed for anti-Crotalus atrox venom antibodies using an enzyme-linked immunosorbent assay (ELISA). Rattlesnake-bitten horses had higher (P = 0.001) titers than vaccinated horses. There was no significant difference between titers in vaccinated pregnant versus nonpregnant horses. One mare had a positive titer at foaling, and the foals had positive postsuckle titers. Antivenom antibody titer development was variable following natural envenomation and vaccination, and vaccine-induced titers were lower than natural envenomation titers. Further studies are required to determine if natural or vaccine antivenom antibody titers reduce the effects of envenomation. PMID:23515015

  20. EphA2 Targeted Chemotherapy Using an Antibody Drug Conjugate in Endometrial Carcinoma

    PubMed Central

    Lee, Jeong-Won; Stone, Rebecca L.; Lee, Sun Joo; Nam, Eun Ji; Roh, Ju-Won; Nick, Alpa M.; Han, Hee-Dong; Shahzad, Mian M.K.; Kim, Hye-Sun; Mangala, Lingegowda S.; Jennings, Nicholas B.; Mao, Shenlan; Gooya, John; Jackson, Dowdy; Coleman, Robert L.; Sood, Anil K.

    2013-01-01

    Purpose EphA2 overexpression is frequently observed in endometrial cancers, and is predictive of poor clinical outcome. Here, we utilize an antibody drug conjugate (MEDI-547) composed of a fully human monoclonal antibody against both human and murine EphA2 (1C1) and the tubulin polymerization inhibitor, monomethylauristatin F (MMAF). Experimental design EphA2 expression was examined in endometrial cancer cell lines by Western Blot. Specificity of MEDI-547 was examined by antibody degradation and internalization assays. Viability and apoptosis were investigated in endometrial cancer cell lines and orthotopic tumor models. Results EphA2 was expressed in the Hec-1A and Ishikawa cells, but was absent in the SPEC-2 cells. Antibody degradation and internalization assays showed that the antibody drug conjugate decreased EphA2 protein levels and was internalized in EphA2 positive cells (Hec-1A and Ishikawa). Moreover, in vitro cytotoxicity and apoptosis assays demonstrated that the antibody drug conjugate decreased viability and increased apoptosis of Hec-1A and Ishikawa cells. In vivo therapy experiments in mouse orthotopic models with this antibody drug conjugate resulted in 86 to 88% growth inhibition (P < 0.001) in the orthotopic Hec-1A and Ishikawa models compared to controls. Moreover, the mice treated with this antibody drug conjugate had a lower incidence of distant metastasis compared with controls. The anti-tumor effects of the therapy were related to decreased proliferation and increased apoptosis of tumor and associated endothelial cells. Conclusions The preclinical data for endometrial cancer treatment using MEDI-547 demonstrate substantial anti-tumor activity. PMID:20388851