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1

Assessment of the rapid test based on an immunochromatography technique for detecting anti-Treponema pallidum antibodies  

Microsoft Academic Search

SUMMARY A rapid test based on an immunochromatography assay - Determine? Syphilis TP (Abbott Lab.) for detecting specific antibodies to Treponema pallidum was evaluated against serum samples from patients with clinical, epidemiological and serological diagnosis of syphilis, patients with sexually transmitted disease other than syphilis, and individuals with negative serology for syphilis. The Determine? test presented the sensitivity of 93.6%,

Neuza Satomi Sato; Carmen Silvia de Melo; Lia C. M. S. Zerbini; Edilene P. R. Silveira; Luiz Jorge Fagundes; Mirthes Ueda

2003-01-01

2

Treponema pallidum Major Sheath Protein Homologue Tpr K Is a Target of Opsonic Antibody and the Protective Immune Response  

Microsoft Academic Search

Summary We have identified a family of genes that code for targets for opsonic antibody and protective immunity in T. pallidum subspecies pallidum using two different approaches, subtraction hybrid- ization and differential immunologic screening of a T. pallidum genomic library. Both ap- proaches led to the identification of a polymorphic multicopy gene family with predicted amino acid homology to the

Arturo Centurion-Lara; Christa Castro; Lynn Barrett; Caroline Cameron; Maryam Mostowfi; Wesley C. Van Voorhis; Sheila A. Lukehart

3

Characterization of the 35-kilodalton Treponema pallidum subsp. pallidum recombinant lipoprotein TmpC and antibody response to lipidated and nonlipidated T. pallidum antigens.  

PubMed Central

The gene encoding the 35-kDa immunogenic Treponema pallidium subsp. pallidum (T. pallidum) membrane protein C, TmpC, was cloned, sequenced, and expressed in Escherichia coli. The deduced amino acid sequence carries on N-terminal signal sequence with a four-amino-acid motif, which is characteristic for bacterial lipoproteins. Metabolic labeling with radioactive palmitic acid of E. coli expressing TmpC revealed incorporation of the fatty acid into the antigen. The antigen was overproduced, purified to near homogeneity and used in an enzyme-linked immunosorbent assay (ELISA) to evaluate its potential for the serodiagnosis of syphilis. Although all sera from untreated secondary syphilis patients were reactive in this TmpC ELISA, only a minority of the serum samples from untreated patients in the primary or early latent stage of the disease contained significant anti-TmpC antibodies. To study the influence of the lipid moiety on the antigenic properties of the TmpC, TmpA, and TpD lipoproteins, plasmids encoding nonlipidated forms of these antigens were constructed. In addition, a plasmid expressing a lipidated form of the otherwise non-lipid-modified antigen TmpB was constructed. Immunization and absorption experiments with these lipidated and nonlipidated antigens showed that antibodies against the lipid moiety of lipoproteins could not be detected on immunoblots, neither in sera from infected rabbits nor in sera from animals immunized with the lipoproteins. In addition, we were unable to demonstrate cross-reactivity between antibodies against the T. pallidum lipoproteins and those reactive to the Venereal Diseases Research Laboratories test, suggesting that antibodies reactive to the Venereal Diseases Research Laboratories test are unrelated to antilipoprotein antibodies. Images PMID:1894360

Schouls, L M; van der Heide, H G; van Embden, J D

1991-01-01

4

Validation of the INNO-LIA Syphilis Kit as a Confirmatory Assay for Treponema pallidum Antibodies  

Microsoft Academic Search

The commercially available diagnostic tests for syphilis are mostly based on the use of extracted antigens of Treponema pallidum. Pronounced cross-reactivities with other spirochete antigens are often reported. The aim of this study was to validate a novel multiparametric assay (the assay performed with the kit) INNO-LIA Syphilis for the confirmation of syphilis antibodies in a set of 840 documented

ANNE EBEL; LIES VANNESTE; MARTINE CARDINAELS; ERWIN SABLON; ISABELLE SAMSON; KATRIEN DE BOSSCHERE; FRANK HULSTAERT; MAAN ZREIN

5

Evaluation of an Enzyme Immunoassay Technique for Detection of Antibodies against Treponema pallidum  

Microsoft Academic Search

In the present study, the performance of an enzyme-linked immunosorbent assay (ELISA) technique (Eti-syphilis-G and Eti-syphilis-M; DiaSorin) for detection of Treponema pallidum immunoglobulin M (IgM) and IgG antibodies for the laboratory diagnosis of syphilis was evaluated. Four hundred forty-one samples were studied. The sensitivity and specificity of the ELISA were 100 and 93%, respectively, compared with the results of a

Rita Castro; Emília S. Prieto; Irene Santo; Jacinta Azevedo; L. Exposto

2003-01-01

6

The TprK Protein of Treponema pallidum Is Periplasmic and Is Not a Target of Opsonic Antibody or Protective Immunity  

Microsoft Academic Search

The finding that Treponema pallidum , the syphilis spirochete, contains 12 orthologs of the Trepo- nema denticola outer membrane major sheath protein has engendered speculation that members of this T. pallidum repeat (Tpr) family may be similarly surface exposed. In this regard, the TprK protein was reported to be a target of opsonic antibody and protective immunity and subject to

Karsten R. O. Hazlett; Timothy J. Sellati; Tung T. Nguyen; David L. Cox; Michael L. Clawson; Melissa J. Caimano; Justin D. Radolf

7

IgG and IgM antibody reactivity to antigens of Treponema pallidum after treatment of syphilis.  

PubMed

The persistence or loss of IgG and IgM antibody specificities for individual polypeptides of Treponema pallidum after therapy for syphilis was examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and by the Western blot technique. Both IgG and IgM antibodies to as many as 12 treponemal antigens, including a major 47-kdalton molecule, were evident in plasma from patients with untreated primary syphilis. IgM reactivity declined rapidly and uniformly after therapy, whereas IgG persisted despite some diminution in intensity of staining. Faint-to-moderate IgM and strong IgG antibody reactivities to at least 22 treponemal antigens (12-85 kdaltons) were identified in plasma from patients with untreated secondary and early latent syphilis. Again, IgG antibody declined slightly in staining intensity after treatment but continued to show reactivity against all molecules detected initially. IgM antibody reactivity declined more rapidly and was lost entirely against some determinants, including the 14- and 12-kdalton molecules. Immunofluorescence titers of IgG and IgM antibodies to T. pallidum in sera from these patients generally correlated with results of Western blot analysis. Antibody to the 12-, 14-, and 47-kdalton molecules of T. pallidum may have potential diagnostic applications. PMID:3544253

Baker-Zander, S A; Roddy, R E; Handsfield, H H; Lukehart, S A

1986-01-01

8

Evaluation of a New Competitive Immunoassay (BioElisa Syphilis) for Screening for Treponema pallidum Antibodies at Various Stages of Syphilis  

Microsoft Academic Search

The BioElisa Syphilis, a new competitive enzyme immunoassay (EIA) for Treponema pallidum whole antigen that uses specific human immunoglobulin G (IgG) antibodies as the competitor, was evaluated for potential use in screening for syphilis at various stages. The results obtained by this competitive EIA were compared with those obtained by the fluorescent treponemal antibody absorption (FTA-abs) test and the T.

ANNE EBEL; LOIC BACHELART; JEAN-MICHEL ALONSO

1998-01-01

9

Expression of Treponema pallidum Antigens in Escherichia coli  

NASA Astrophysics Data System (ADS)

Treponema pallidum DNA was cloned in a bacteriophage. Clones were screened for expression of Treponema pallidum antigens by an in situ radio-immunoassay on nitrocellulose, with the use of subsequent reactions with syphilitic serum and radioiodinated Staphylococcus aureus protein A. One clone, which gave a strong signal, codes for at least seven antigens that react specifically with human antibodies to Treponema pallidum.

Walfield, Alan M.; Hanff, Philip A.; Lovett, Michael A.

1982-04-01

10

Protection against Syphilis Correlates with Specificity of Antibodies to the Variable Regions of Treponema pallidum Repeat Protein K  

Microsoft Academic Search

Syphilis has been recognized as a disease since the late 1400s, yet there is no practical vaccine available. One impediment to the development of a vaccine is the lack of understanding of multiple reinfections in humans despite the development of robust immune responses during the first episode. It has been shown that the Treponema pallidum repeat protein K (TprK) differs

Cecilia A. Morgan; Sheila A. Lukehart; Wesley C. Van Voorhis

2003-01-01

11

Treponema pallidum Surface Immunofluorescence Assay for Serologic Diagnosis of Syphilis  

Microsoft Academic Search

A surface immunofluorescence assay (SIFA) using live spirochetes was analyzed and compared with Western blot (WB), fluorescent treponemal antibody absorption (FTA-ABS), microhemagglutination (MHA-TP), and Treponema pallidum immobilization (TPI) assays for detecting serum antibodies to T. pallidum in patients with syphilis, in disease controls, and in healthy subjects. SIFA and WB were 99% sensitive (99 of 100 positive specimens) and specific

ANTONELLA MARANGONI; VITTORIO SAMBRI; ELISA STORNI; A. D'Antuono; M. Negosanti; R. Cevenini

2000-01-01

12

Comparative Evaluation of Nine Different Enzyme-Linked Immunosorbent Assays for Determination of Antibodies against Treponema pallidum in Patients with Primary Syphilis  

Microsoft Academic Search

Nine different enzyme-linked immunosorbent assays (ELISAs) with a sonicate or recombinant proteins of Treponema pallidum as antigen have been evaluated comparatively by testing 52 highly selected sera from patients with primary syphilis, all negative in the microhemagglutination test for T. pallidum (MHA-TP). Eight tests exhibited greater sensitivity (48.5 to 76.9%) than the commonly used Venereal Disease Research Labo- ratory test

BRUNO L. SCHMIDT; MARZIEH EDJLALIPOUR; ANTON LUGER; Ludwig Boltzmann

2000-01-01

13

Identification of the Treponema pallidum subsp. pallidum glycerophosphodiester phosphodiesterase homologue  

Microsoft Academic Search

To identify potential opsonic targets of Treponema pallidum subsp. pallidum, a treponemal genomic expression library was constructed and differentially screened with opsonic and non-opsonic T. pallidum antisera. This method identified an immunoreactive clone containing an open reading frame encoding a 356 residue protein. Nucleotide sequence analysis demonstrated the translated protein to be a homologue of glycerophosphodiester phosphodiesterase, a glycerol metabolizing

Caroline E. Stebeck; Jeanne M. Shaffer; Thomas W. Arroll; Sheila A. Lukehart; Wesley C. Van Voorhis

1997-01-01

14

The axial filament antigen of Treponema pallidum.  

PubMed Central

Axial filaments (flagella) of Treponema pallidum have been purified in large enough quantities to be analysed electrophoretically. They produced a characteristic linear precipitate in two-dimensional immunoelectrophoresis. Polyacrylamide gel electrophoresis showed three major polypeptides, the most prominent having an apparent molecular weight of 37,000, about 1500 less than the corresponding component of axial filaments of the Reiter treponeme. A doublet of less abundant polypeptides of 33,500-34,000 MW also differed slightly from those of the Reiter treponeme. Western blot analysis showed that the principal polypeptide of the T. pallidum axial filament was strongly antigenic, and antibody to it was prominent in sera from hyperimmunized, as well as acutely infected (orchitic), rabbits, and in soluble fractions from acutely infected rabbit testes from which large numbers of viable treponemes had been extracted. This indicated that antibody to this component was ineffective in eliminating treponemes from the tissue. Images Figure 1 Figure 2 Figure 3 Figure 4 PMID:3884491

Penn, C W; Bailey, M J; Cockayne, A

1985-01-01

15

The Treponema pallidum Genome Database  

NSDL National Science Digital Library

The Institute for Genomic Research (TIGR) has posted the complete gene sequence of Treponema pallidum (published in Science 281:375-388, 1998). The newly available Treponema pallidum genome, directed by Dr. Claire Fraser, is searchable by name, TP number, sequence, and segment. Offerings at the site include a hyperlinked Gene Identification Table, RNA Gene Table, Paralogous gene families of Treponema pallidum, and instructions on how to download data.

Fraser, Claire.

1998-01-01

16

Detection of Treponema pallidum by a Sensitive Reverse Transcriptase PCR  

Microsoft Academic Search

Syphilis is diagnosed by serologic testing or by identification of the causative agent, Treponema pallidum. The bacterium has historically been detected in clinical specimens by dark-field microscopy, immunostaining with polyclonal or monoclonal antibodies, or the rabbit inoculation test (RIT). RIT is considered to be very sensitive and specific, although it is available only in research settings and is not clinically

ARTURO CENTURION-LARA; CHRISTA CASTRO; JEANNE M. SHAFFER; WESLEY C. VAN VOORHIS; CHRISTINA M. MARRA; SHEILA A. LUKEHART

1997-01-01

17

Recombinant treponema pallidum antigens in syphilis serology  

Microsoft Academic Search

Treponema pallidum, the etiological agent of syphilis, is characterized by a paucity of surface exposed outer membrane proteins and a high content of cytoplasma membrane associated lipoproteins. At all stages of infection intense antibody responses against lipoproteins are detectable. In order to provide antigens for syphilis diagnosis the highly immunogenic lipoproteins TpN17, TpN29-35 (TpD), TpN44.5 (TmpA), TpN47, and TpN35 (TmpC)

Annegret Gerber; Siegfried Krell; Joachim Morenz

1997-01-01

18

Plasmonic ELISA for the ultrasensitive detection of Treponema pallidum.  

PubMed

In this report, we have developed a plasmonic ELISA strategy for the detection of syphilis. Plasmonic ELISA is an enzyme-linked immunoassay combined with enzyme-mediated surface plasmon resonance (SPR) of gold nanoparticles (AuNPs). Immune response of the Treponema pallidum (T. pallidum) antibodies triggers the acetylcholinesterase-catalyzed hydrolysis of acetylthiocholine to produce abundant thiocholine. The positive charged thiol, in turn, alters the surface charge distribution the AuNPs and leads to the agglomeration of the AuNPs. The induced strong localized SPR effect of the agglomerate AuNPs can, thus, allow the quantitative assay of T. pallidum antibodies due to the remarkable color and absorption spectral response changes of the reaction system. The plasmonic ELISA exhibited a quasilinear response to the logarithmic T. pallidum antibody concentrations in the range of 1pg/mL-10ng/mL with a detection limit of 0.98pg/mL. Such a low detection limit was 1000-fold improvements in sensitivity over a conventional ELISA. The results of plasmonic ELISA in syphilis assays of serum specimens from 60 patients agreed with those obtained using a conventional ELISA method. The plasmonic ELISA has characteristics (analyte specific, cost-effective, ease of automatic, low limit of detection) that provide potential for diagnosis and therapeutic monitoring of syphilis. PMID:24662060

Nie, Xin-Min; Huang, Rong; Dong, Cai-Xia; Tang, Li-Juan; Gui, Rong; Jiang, Jian-Hui

2014-08-15

19

Function and Protective Capacity of Treponema pallidum subsp. pallidum Glycerophosphodiester Phosphodiesterase  

Microsoft Academic Search

Infectious syphilis, caused by the spirochete bacterium Treponema pallidum subsp. pallidum, remains a public health concern worldwide. The immune-response evasion mechanisms employed by T. pallidum are poorly under- stood, and prior attempts to identify immunoprotective antigens for subsequent vaccine design have been unsuc- cessful. Previous investigations conducted in our laboratory identified the T. pallidum glycerophosphodiester phosphodiesterase as a potential immunoprotective

CAROLINE E. CAMERON; CHRISTA CASTRO; SHEILA A. LUKEHART; WESLEY C. VAN VOORHIS

1998-01-01

20

Clinical Comparison of the Treponema pallidum CAPTIA Syphilis-G Enzyme Immunoassay with the Fluorescent Treponemal Antibody Absorption Immunoglobulin G Assay for Syphilis Testing  

Microsoft Academic Search

Recently, a treponema-specific immunoglobulin G (IgG) enzyme immunoassay (EIA), the CAPTIA Syphi- lis-G (Trinity Biotech, Jamestown, N.Y.), has become available as a diagnostic test for syphilis. A total of 89 stored sera previously tested by the fluorescent treponemal antibody absorption (FTA-ABS) IgG assay were evaluated by the CAPTIA EIA. The FTA-ABS IgG procedure was performed by technologists unblinded to results

V. W. HALLING; M. F. JONES; J. E. BESTROM; A. D. WOLD; J. E. ROSENBLATT; T. F. SMITH; F. R. COCKERILL

1999-01-01

21

Relationship between neurological features and intrathecal synthesis of IgG antibodies to Treponema pallidum in untreated and treated human neurosyphilis  

Microsoft Academic Search

In syphilitic patients with or without CNS involvement the correlation of Treponema-specific IgG titre per milligram total IgG in CSF and serum (ITPA index) is a dependable source of information on the synthesis of treponemal IgG antibodies in the CNS. This index also provides a more reliable definition of asymptomatic neurosyphilis. Further, a discrimination between Treponema-specific and Treponema-non-specific IgG synthesis

H. W. Prange; M. Moskophidis; H. I. Schipper; F. Müller

1983-01-01

22

Compositions and Methods for Detecting Treponema Pallidum.  

National Technical Information Service (NTIS)

Methods for the specific and highly sensitive detection of Treponema pallidum infection comprising the use of specific antigenic proteins and peptides unique to Treponema pallidum are provided. In particular, detection assays based recognition of acidic r...

B. Rodes, B. M. Steiner, H. Liu

2001-01-01

23

The tprK Gene Is Heterogeneous among Treponema pallidum Strains and Has Multiple Alleles  

Microsoft Academic Search

We have previously shown that the TprK antigen of T. pallidum, Nichols strain, is predominantly expressed in treponemes obtained 10 days after infection and that the hydrophilic domain of TprK is a target of opsonic antibodies and confers significant protection against homologous challenge. The T. pallidum genome sequence reported the presence of a single copy of the tprK gene in

ARTURO CENTURION-LARA; CHARMIE GODORNES; CHRISTA CASTRO; WESLEY C. VAN VOORHIS; SHEILA A. LUKEHART

2000-01-01

24

Ventral Pallidum Roles in Reward and Motivation  

PubMed Central

In recent years the ventral pallidum has become a focus of great research interest as a mechanism of reward and incentive motivation. As a major output for limbic signals, the ventral pallidum was once associated primarily with motor functions rather than regarded as a reward structure in its own right. However, ample evidence now suggests that ventral pallidum function is a major mechanism of reward in the brain. We review data indicating that 1) an intact ventral pallidum is necessary for normal reward and motivation, 2) stimulated activation of ventral pallidum is sufficient to cause reward and motivation enhancements, and 3) activation patterns in ventral pallidum neurons specifically encode reward and motivation signals via phasic bursts of excitation to incentive and hedonic stimuli. We conclude that the ventral pallidum may serve as an important ‘limbic final common pathway’ for mesocorticolimbic processing of many rewards. PMID:18955088

Smith, Kyle S.; Tindell, Amy J.; Aldridge, J. Wayne; Berridge, Kent C.

2008-01-01

25

Native surface association of a recombinant 38-kilodalton Treponema pallidum antigen isolated from the Escherichia coli outer membrane.  

PubMed Central

A recombinant plasmid designated pAW305, containing a 6-kilobase insert of Treponema pallidum DNA, directed the expression of a 38-kilodalton (kDa) treponemal antigen in Escherichia coli. The 38-kDa antigen copurified with the outer membrane fraction of the E. coli cell envelope after treatment with nonionic detergents or sucrose density gradient centrifugation. Rabbits immunized with the recombinant 38-kDa antigen developed antibodies which reacted specifically with a 38-kDa T. pallidum antigen on immunoblots, and 38-kDa antisera specifically immobilized T. pallidum in a complement-dependent manner in the T. pallidum immobilization test. Antisera to the 38-kDa recombinant antigen were also used to demonstrate its native surface association on T. pallidum by immunoelectron microscopy. Images PMID:3516880

Fehniger, T E; Radolf, J D; Walfield, A M; Cunningham, T M; Miller, J N; Lovett, M A

1986-01-01

26

Polypeptides of Treponema pallidum: progress toward understanding their structural, functional, and immunologic roles. Treponema Pallidum Polypeptide Research Group.  

PubMed Central

Treponema pallidum subsp. pallidum, the spirochete that causes syphilis, is unusual in a number of respects, including its small genome size, inability to grow under standard in vitro culture conditions, microaerophilism, apparent paucity of outer membrane proteins, structurally complex periplasmic flagella, and ability to evade the host immune responses and cause disease over a period of years to decades. Many of these attributes are related ultimately to its protein content. Our knowledge of the activities, structure, and immunogenicity of its proteins has been expanded by the application of recombinant DNA, hybridoma, and structural fractionation techniques. The purpose of this monograph is to summarize and correlate this new information by using two-dimensional gel electrophoresis, monoclonal antibody reactivity, sequence data, and other properties as the bases of polypeptide identification. The protein profiles of the T. pallidum subspecies causing syphilis, yaws, and endemic syphilis are virtually indistinguishable but differ considerably from those of other treponemal species. Among the most abundant polypeptides are a group of lipoproteins of unknown function that appear to be important in the immune response during syphilitic infection. The periplasmic flagella of T. pallidum and other spirochetes are unique with regard to their protein content and ultrastructure, as well as their periplasmic location. They are composed of three core proteins (homologous to the other members of the eubacterial flagellin family) and a single, unrelated sheath protein; the functional significance of this arrangement is not understood at present. Although the bacterium contains the chaperonins GroEL and DnaK, these proteins are not under the control of the heat shock regulon as they are in most organisms. Studies of the immunogenicity of T. pallidum proteins indicate that many may be useful for immunodiagnosis and immunoprotection. Future goals in T. pallidum polypeptide research include continued elucidation of their structural locations and functional activities, identification and characterization of the low-abundance outer membrane proteins, further study of the immunoprotective and immunodiagnostic potential of T. pallidum proteins, and clarification of the roles of treponemal proteins in pathogenesis. Images PMID:8246847

Norris, S J

1993-01-01

27

Multiple Alleles of Treponema pallidum Repeat Gene D in Treponema pallidum Isolates  

Microsoft Academic Search

Syphilis, caused by Treponema pallidum subsp. pallidum ,i s a chronic disease characterized by periods of activity and latency, with clearance of early lesions but persistence of infection. The mechanisms T. pallidum uses to persist in humans are still not known, but it is possible that alterations in surface proteins will be involved because of their interaction with the host

ARTURO CENTURION-LARA; EILEEN S. SUN; LYNN K. BARRETT; CHRISTA CASTRO; SHEILA A. LUKEHART; WESLEY C. VAN VOORHIS

2000-01-01

28

Treponema pallidum western blot: Comparison with the FTA-ABS test as a confirmatory test for syphilis  

Microsoft Academic Search

We developed a Treponema pallidum Western blot and compared the results with Treponema pallidum particle agglutination (TPPA) and fluorescent treponemal antibody absorption (FTA-ABS) tests. The Western blot was deemed reactive if the serum reacted with at least three major antigenic bands (TpN47, TpN44.5, TpN17, TpN15). The sensitivities of the Western blot, TPPA and FTA-ABS, were all 100% and the specificities

Josephine L. Backhouse; Serge I. Nesteroff

2001-01-01

29

Major integral membrane protein immunogens of Treponema pallidum are proteolipids  

SciTech Connect

A number of the major pathogen-specific immunogens of Treponema pallidum were characterized recently as amphiphilic, integral membrane proteins by phase partitioning with Triton X-114. In the present study, we demonstrated that the same membrane immunogens (designated as detergent phase proteins (DPPs)) become radiolabeled upon in vitro incubation of T. pallidum with various {sup 3}H-labeled fatty acids. Radioimmunoprecipitation with a monoclonal antibody confirmed that the {sup 3}H-labeled 47-kilodalton protein corresponded to the well-characterized treponemal antigen with the identical apparent molecular mass. Failure to detect {sup 3}H-labeled DPPs following incubation with erythromycin confirmed that protein acylation required de novo protein synthesis by the bacteria. When treponemes were incubated with ({sup 3}H)myristate, ({sup 3}H)palmitate, or ({sup 3}H)oleate, radiolabeled proteins corresponding to the DPPs were detected upon autoradiography. Demonstration that a number of the abundant membrane immunogens of T. pallidum are proteolipids provides information to help clarify their membrane association(s) and may serve to explain their extraordinary immunogenicity.

Chamberlain, N.R.; Brandt, M.E.; Erwin, A.L.; Radolf, J.D.; Norgard, M.V. (Univ. of Texas Southwestern Medical Center, Dallas (USA))

1989-09-01

30

Purification and characterization of a cloned protease-resistant Treponema pallidum-specific antigen.  

PubMed Central

A cloned Treponema pallidum antigen, designated 4D, was purified from Escherichia coli predominantly as a 190-kilodalton (kd) polypeptide, although higher oligomeric forms exist. Extensive proteolysis of 4D created a limit digestion product of 90 kd which retained antigenicity with sera from patients with primary, secondary, early latent, late latent, and tertiary syphilis. A molecule indistinguishable from 90-kd 4D in size, isoelectric point, and antigenicity was isolated from T. pallidum after proteolysis. The 190- and 90-kd forms of 4D were stable at 68 degrees C but converted to 19- and 14-kd species, respectively, after boiling in sodium dodecyl sulfate. The low-molecular-weight species did not react with syphilitic sera. Rabbits immunized with the purified 4D antigen developed antibodies which immobilized virulent T. pallidum in a complement-dependent assay system, suggesting that the antigen has a native surface location. Images PMID:6389353

Fehniger, T E; Walfield, A M; Cunningham, T M; Radolf, J D; Miller, J N; Lovett, M A

1984-01-01

31

The Treponema pallidum immobilization test  

PubMed Central

The authors first discuss the principle of the Treponema pallidum immobilization (TPI) test, considered both as a qualitative and as a quantitative test, and then various specific aspects of passage of the Nichols strain of treponeme in rabbit testes. A number of important technical details and modifications in the test procedure are then reviewed, and the clinical value of the test is discussed. The sensitivity of the TPI test is considerably greater than that of the serological tests for syphilis more usually performed, and its specificity is also remarkably high. However, it is stressed that, at the present stage of development, the greatest value of the TPI test lies in its use as an excellent aid to diagnosis rather than as a test of cure. PMID:13329850

Nielsen, Hans Aage; Reyn, Alice

1956-01-01

32

Cultivation of 'Treponema pallidum in vitro'.  

National Technical Information Service (NTIS)

Viability of Treponema pallidum in vitro has been improved using supplemental amino acids, carbohydrates, fatty acids, vitamins and reducing agents in the medium and low levels of oxygen in the cultures. Virulent microorganisms were detected at least six ...

H. M. Jenkin, P. L. Sandok

1977-01-01

33

Complete Genome Sequence of the Treponema pallidum subsp. pallidum Sea81-4 Strain  

PubMed Central

Using the rabbit model of syphilis, the Sea81-4 strain of Treponema pallidum subsp. pallidum has been found to be more likely than other strains to invade the central nervous system (CNS). To identify possible explanations for this important phenotype at the genomic level, we sequenced the Sea81-4 strain genome. PMID:24744342

Iverson-Cabral, Stefanie L.; King, Jordon C. K.; Molini, Barbara J.; Lukehart, Sheila A.; Centurion-Lara, Arturo

2014-01-01

34

Complete Genome Sequence of the Treponema pallidum subsp. pallidum Sea81-4 Strain.  

PubMed

Using the rabbit model of syphilis, the Sea81-4 strain of Treponema pallidum subsp. pallidum has been found to be more likely than other strains to invade the central nervous system (CNS). To identify possible explanations for this important phenotype at the genomic level, we sequenced the Sea81-4 strain genome. PMID:24744342

Giacani, Lorenzo; Iverson-Cabral, Stefanie L; King, Jordon C K; Molini, Barbara J; Lukehart, Sheila A; Centurion-Lara, Arturo

2014-01-01

35

Osteitis in the dens of axis caused by Treponema pallidum  

PubMed Central

Background Syphilis has been referred to as “the great imitator” due to its ability to imitate other diseases. Untreated syphilis becomes a systemic infection that can involve almost every organ systems. Treponema pallidum has a high affinity for bone tissue, but osteitis has mainly been described in late stages of the disease. Vertebral involvement is rare, and this is to our knowledge the first case describing syphilitic spondylitis in early acquired syphilis. Case presentation We here describe destructive osteitis in the vertebral column as the initial manifestation of early acquired syphilis in a 24-year-old caucasian homosexual male with HIV infection. The diagnosis was reached by universal bacterial PCR and DNA sequencing of the DNA product. It was confirmed by PCR specific for Treponema pallidum, immunohistochemistry and detection of increasing antibody titer. Conclusions As syphilis has re-emerged in Western countries and remains a worldwide common disease it is important to have in mind as a causative agent of skeletal symptoms, especially among HIV-infected individuals or men who have sex with men (MSM). PMID:23885957

2013-01-01

36

Effect of pretreatment with Mycobacterium bovis (strain BCG) and immune syphilitic serum on rabbit resistance to Treponema pallidum.  

PubMed Central

Stimulation of the rabbit reticuloendothelial system with viable Mycobacterium bovis (strain BCG), and other agents, had no effect on the development of syphilitic lesions after intradermal or intravenous inoculation with graded doses of Treponema pallidum (virulent Nichol's strain; mean infective doses less than 10). The simultaneous administration of immune syphilitic rabbit serum retarded the development of lesions, but this appeared to be due solely to the immune serum, suggesting no synergism between the activated reticuloendothelial system and the anti-T. pallidum antibodies. The administration of two doses of BCG enhanced syphilitic lesion development in the rabbit. PMID:172450

Graves, S R; Johnson, R C

1975-01-01

37

Cultivation of 'Treponema pallidum' 'in vitro'.  

National Technical Information Service (NTIS)

The authors have increased the retention of active motility of Treponema pallidum from 3-4 days to 8 or more days as well as maintaining virulence of the organism by 6-7 days by improving the media and environmental conditions. These results were obtained...

H. M. Jenkin

1976-01-01

38

Cultivation of 'Treponema pallidum' in vitro.  

National Technical Information Service (NTIS)

Prereduced Eagle's minimal essential medium (PRMEM10) has a factor(s) in it which enhances the maintenance of motility of T. pallidum virulent Nichols I (T. pall. vi. Ni.) under anaerobic conditions. Preliminary results suggest that PRMEM10 mixed with spe...

H. M. Jenkin

1975-01-01

39

21 CFR 866.3830 - Treponema pallidum tre-ponemal test reagents.  

Code of Federal Regulations, 2010 CFR

... 2010-04-01 false Treponema pallidum tre-ponemal test reagents...Serological Reagents § 866.3830 Treponema pallidum tre-ponemal test reagents. (a) Identification. Treponema pallidum treponemal test...

2010-04-01

40

21 CFR 866.3820 - Treponema pallidum non-treponemal test reagents.  

Code of Federal Regulations, 2012 CFR

... 2012-04-01 false Treponema pallidum non-treponemal test reagents...Serological Reagents § 866.3820 Treponema pallidum non-treponemal test reagents. (a) Identification. Treponema pallidum nontreponemal...

2012-04-01

41

21 CFR 866.3830 - Treponema pallidum tre-ponemal test reagents.  

... 2014-04-01 false Treponema pallidum tre-ponemal test reagents...Serological Reagents § 866.3830 Treponema pallidum tre-ponemal test reagents. (a) Identification. Treponema pallidum treponemal test...

2014-04-01

42

21 CFR 866.3820 - Treponema pallidum non-treponemal test reagents.  

Code of Federal Regulations, 2013 CFR

... 2013-04-01 false Treponema pallidum non-treponemal test reagents...Serological Reagents § 866.3820 Treponema pallidum non-treponemal test reagents. (a) Identification. Treponema pallidum nontreponemal...

2013-04-01

43

21 CFR 866.3820 - Treponema pallidum non-treponemal test reagents.  

Code of Federal Regulations, 2011 CFR

... 2011-04-01 false Treponema pallidum non-treponemal test reagents...Serological Reagents § 866.3820 Treponema pallidum non-treponemal test reagents. (a) Identification. Treponema pallidum nontreponemal...

2011-04-01

44

Treponema pallidum receptor binding proteins interact with fibronectin  

SciTech Connect

Analysis of plasma proteins avidly bound to T. pallidum surfaces revealed the ability of T. pallidum to acquire numerous host macromolecules. No acquisition was evident by the avirulent spirochete, T. phagedenis biotype Reiter. Western blotting technology using hyperimmune antifibronectin serum as a probe revealed the ability of virulent treponemes to avidly bind fibronectin from a complex medium such as plasma. The specificity of the tiplike adherence of motile T. pallidum to fibronectin-coated glass surfaces and to fibronectin on HEp-2 cells was reinforced by the observation that pretreatment of coverslips or cell monolayers with monospecific antiserum against fibronectin substantially reduced T. pallidum attachment. The stoichiometric binding of T. pallidum to fibronectin-coated coverslips and the inability of unlabeled or /sup 35/S-radiolabeled treponemes to interact with glass surfaces treated with other plasma proteins further established the specific nature of the interaction between virulent T. pallidum and fibronectin. The avid association between three outer envelope proteins of T. pallidum and fibronectin was also demonstrated. These treponemal surface proteins have been previously identified as putative receptor-binding proteins responsible for T. pallidum parasitism of host cells. The data suggest that surface fibronectin mediates tip-oriented attachment of T. pallidum to host cells via a receptor-ligand mechanism of recognition.

Peterson, K.M.; Baseman, J.B.; Alderete, J.F.

1983-06-01

45

TP0262 is a modulator of promoter activity of tpr Subfamily II genes of Treponema pallidum ssp. pallidum  

Microsoft Academic Search

Summary Transcriptional regulation in Treponema pallidum ssp. pallidum is poorly understood, primarily because this organism cannot be cultivated in vitro or geneti- cally manipulated. We have recently shown a phase variation mechanism controlling transcription initia- tion of Subfamily II tpr (T. pallidum repeat) genes (tprE, tprG and tprJ), a group of virulence factor candidates. Furthermore, the same study suggested that

Lorenzo Giacani; Charmie Godornes; Maritza Puray-Chavez; Cristina Guerra-Giraldez; Martin Tompa; Sheila A. Lukehart; Arturo Centurion-Lara

2009-01-01

46

Sequence Diversity of Treponema pallidum subsp. pallidum tprK in Human Syphilis Lesions and Rabbit-Propagated Isolates  

Microsoft Academic Search

The tprK gene of Treponema pallidum subsp. pallidum, the causative agent of venereal syphilis, belongs to a 12-member gene family and encodes a protein with a predicted cleavable signal sequence and predicted transmembrane domains. Except for the Nichols type strain, all rabbit-propagated isolates of T. pallidum examined thus far are comprised of mixed populations of organisms with heterogeneous tprK sequences.

Rebecca E. LaFond; Arturo Centurion-Lara; Charmie Godornes; Anne M. Rompalo; Wesley C. Van Voorhis; Sheila A. Lukehart

2003-01-01

47

Treponema pallidum receptor binding proteins interact with fibronectin  

Microsoft Academic Search

Analysis of plasma proteins avidly bound to T. pallidum surfaces revealed the ability of T. pallidum to acquire numerous host macromolecules. No acquisition was evident by the avirulent spirochete, T. phagedenis biotype Reiter. Western blotting technology using hyperimmune antifibronectin serum as a probe revealed the ability of virulent treponemes to avidly bind fibronectin from a complex medium such as plasma.

KENNETH M. PETERSON; JOEL B. BASEMAN; JOHN F. ALDERETE

1983-01-01

48

Treponema pallidum Invades Intercellular Junctions of Endothelial Cell Monolayers  

Microsoft Academic Search

The pathogenesis of syphilis reflects invasive properties of Treponema pallidum, but the actual mode of tissue invasion is unknown. We have found two in vitro parallels of treponemal invasiveness. We tested whether motile T. pallidum could invade host cells by determining the fate of radiolabeled motile organisms added to a HeLa cell monolayer; 26% of treponemes associated with the monolayer

D. Denee Thomas; Mahamad Navab; David A. Haake; Alan M. Fogelman; James N. Miller; Michael A. Lovett

1988-01-01

49

Antibody  

MedlinePLUS

An antibody is a protein produced by the body's immune system when it detects harmful substances, called antigens. Examples ... as bacteria, fungi, parasites, and viruses) and chemicals. Antibodies may be produced when the immune system mistakenly ...

50

Adjuvant activity of Sargassum pallidum polysaccharides against combined Newcastle disease, infectious bronchitis and avian influenza inactivated vaccines.  

PubMed

This study evaluates the effects of Sargassum pallidum polysaccharides (SPP) on the immune responses in a chicken model. The adjuvanticity of Sargassum pallidum polysaccharides in Newcastle disease (ND), infectious bronchitis (IB) and avian influenza (AI) was investigated by examining the antibody titers and lymphocyte proliferation following immunization in chickens. The chickens were administrated combined ND, IB and AI inactivated vaccines containing SPP at 10, 30 and 50 mg/mL, using an oil adjuvant vaccine as a control. The ND, IB and AI antibody titers and the lymphocyte proliferation were enhanced at 30 mg/mL SPP. In conclusion, an appropriate dose of SPP may be a safe and efficacious immune stimulator candidate that is suitable for vaccines to produce early and persistent prophylaxis. PMID:23342387

Li, Li-Jie; Li, Ming-Yi; Li, Yan-Tuan; Feng, Jing-Jing; Hao, Feng-Qiang; Lun, Zhang

2012-12-01

51

Antibody  

NSDL National Science Digital Library

Antibody: A blood protein that is produced in response to and counteracts an antigen. Antibodies are produced in response to disease and help the body fight against the particular disease. In this way, antibodies help the body develop an immunity to disease.

Darryl Leja (National Human Genome Research Institute REV)

2005-04-04

52

Flagellins, but Not Endoflagellar Sheath Proteins, of Treponema pallidum and of Pathogen-Related Oral Spirochetes Are Glycosylated  

Microsoft Academic Search

Glycosylation of the flagellar core proteins (FlaBs) was detected in Treponema pallidum Nichols and in the type or reference strains of seven oral Treponema species. In several nonmotile strains of oral treponemes, the FlaBs were undetectable by both antibody and glycan staining. In contrast, a spontaneous low-motility variant of T. vincentiŁi-related strain RitzA, OMZ 305A, lacked the flagellar sheath protein

C. WYSS

53

Treponema pallidum Invades Intercellular Junctions of Endothelial Cell Monolayers  

NASA Astrophysics Data System (ADS)

The pathogenesis of syphilis reflects invasive properties of Treponema pallidum, but the actual mode of tissue invasion is unknown. We have found two in vitro parallels of treponemal invasiveness. We tested whether motile T. pallidum could invade host cells by determining the fate of radiolabeled motile organisms added to a HeLa cell monolayer; 26% of treponemes associated with the monolayer in a trypsin-resistant niche, presumably between the monolayer and the surface to which it adhered, but did not attain intracellularity. Attachment of T. pallidum to cultured human and rabbit aortic and human umbilical vein endothelial cells was 2-fold greater than to HeLa cells. We added T. pallidum to aortic endothelial cells grown on membrane filters under conditions in which tight intercellular junctions had formed. T. pallidum was able to pass through the endothelial cell monolayers without altering tight junctions, as measured by electrical resistance. In contrast, heat-killed T. pallidum and the nonpathogen Treponema phagedenis biotype Reiter failed to penetrate the monolayer. Transmission electron micrographs of sections of the monolayer showed T. pallidum in intercellular junctions. Our in vitro observations suggest that these highly motile spirochetes may leave the circulation by invading the junctions between endothelial cells.

Thomas, D. Denee; Navab, Mahamad; Haake, David A.; Fogelman, Alan M.; Miller, James N.; Lovett, Michael A.

1988-05-01

54

Similarity between the 38-kilodalton lipoprotein of Treponema pallidum and the glucose/galactose-binding (MglB) protein of Escherichia coli.  

PubMed Central

The recent discovery that abundant and immunogenic lipoproteins constitute the integral membrane proteins of Treponema pallidum has prompted efforts to investigate their importance in the physiology and ultrastructure of the organism and in immune responses during infection. Earlier studies identified a 38-kDa lipoprotein of T. pallidum believed to be specific to the pathogen. In the present study, monoclonal antibodies generated against the 38-kDa lipoprotein of T. pallidum reacted with cognate 37-kDa molecules in the nonpathogens Treponema phagedenis, Treponema denticola, and Treponema refringens. Cloning and expression of the 38-kDa-lipoprotein gene of T. pallidum in Escherichia coli revealed that the recombinant product displayed a slightly larger (39-kDa) apparent molecular mass but remained reactive with anti-38-kDa-protein monoclonal antibodies. The recombinant product was processed and acylated in E. coli. DNA and amino acid sequence analyses indicated an open reading frame encoding 403 amino acids, with the first 25 amino acids corresponding to a leader peptide terminated by a signal peptidase II processing site of Val-Val-Gly-Cys. The predicted mature protein is 378 amino acids in length with a deduced molecular weight of 40,422 (excluding acylation). Southern blotting failed to demonstrate in nonpathogenic treponemes genomic sequences homologous with the 38-kDa-lipoprotein gene of T. pallidum. Computer analysis revealed that the 38-kDa lipoprotein of T. pallidum had 34.2% identity and 58.9% similarity with the glucose/galactose-binding protein (MglB) of E. coli and Salmonella typhimurium. Furthermore, of the 19 amino acids of MglB involved in carbohydrate binding, the 38-kDa lipoprotein had identity with 11. These studies have allowed the first putative functional assignment (carbohydrate binding) to a T. pallidum integral membrane protein. Recognition of this potential physiological role for the 38-kDa lipoprotein underscores the possibility that the membrane biology of T. pallidum may more closely resemble that of gram-positive organisms, which also utilize lipoproteins as anchored transporters, than that of gram-negative bacteria to which T. pallidum often is analogized. Images PMID:8132345

Becker, P S; Akins, D R; Radolf, J D; Norgard, M V

1994-01-01

55

Complete genome sequence of Treponema pallidum strain DAL-1  

PubMed Central

Treponema pallidum strain DAL-1 is a human uncultivable pathogen causing the sexually transmitted disease syphilis. Strain DAL-1 was isolated from the amniotic fluid of a pregnant woman in the secondary stage of syphilis. Here we describe the 1,139,971 bp long genome of T. pallidum strain DAL-1 which was sequenced using two independent sequencing methods (454 pyrosequencing and Illumina). In rabbits, strain DAL-1 replicated better than the T. pallidum strain Nichols. The comparison of the complete DAL-1 genome sequence with the Nichols sequence revealed a list of genetic differences that are potentially responsible for the increased rabbit virulence of the DAL-1 strain. PMID:23449808

Zobanikova, Marie; Mikolka, Pavol; Cejkova, Darina; Pospisilova, Petra; Chen, Lei; Strouhal, Michal; Qin, Xiang; Weinstock, George M.; Smajs, David

2012-01-01

56

Penicillin-binding proteins and peptidoglycan of Treponema pallidum subsp. pallidum.  

PubMed Central

Penicillin-binding proteins (PBPs) of Treponema pallidum subsp. pallidum (T. pallidum) were characterized by using [3H]penicillin G and a conjugate consisting of ampicillin and 125I-labeled Bolton-Hunter reagent. Both antibiotics specifically radiolabeled proteins with molecular masses of 94, 80, 63, and 58 kilodaltons (kDa); 125I-labeled Bolton-Hunter reagent-ampicillin also radiolabeled several polypeptides with lower molecular masses. The 94- and 58-kDa proteins demonstrated the highest binding affinities for [3H]penicillin G and were radiolabeled at concentrations of 8 and 40 nM, respectively. Radiolabeling of PBPs was detectable after 1 min of incubation in 1 microM [3H]penicillin G and was nearly maximal within 10 min. The rapidity of penicillin binding contrasted with the observation that only 40% of virulent treponemes became immobilized during prolonged incubation in vitro with a much higher concentration (1 mM) of unlabeled penicillin. Two lines of evidence indicated that most, if not all, of the PBPs are integral cytoplasmic membrane proteins: (i) preincubation of organisms in 0.1% Triton X-100 solubilized nearly all of the outer membranes but did not affect radiolabeling of PBPs, and (ii) except for the 80-kDa protein, the PBPs partitioned into the detergent phase following extraction with the nonionic detergent Triton X-114. The presence of peptidoglycan in T. pallidum was confirmed by the detection of muramic acid in the sodium dodecyl sulfate-insoluble, proteinase K-resistant residue obtained from Triton X-114-extracted organisms. Images PMID:2647634

Radolf, J D; Moomaw, C; Slaughter, C A; Norgard, M V

1989-01-01

57

Cell-Wall Defective Variants of 'Treponema pallidum'.  

National Technical Information Service (NTIS)

Ultrastructural study of selected nonpathogenic strains of T. pallidum (Kazan 2, Kazan 5, Kazan 8 and Reiter) and the pathogenic Nichols strain permitted visualization of the cell wall, cytoplasm, and fibrils of these organisms. Induction of morphological...

M. M. Rodrigues

1972-01-01

58

Effects of Various Handling and Storage Conditions on Stability of Treponema pallidum DNA in Cerebrospinal Fluid  

Microsoft Academic Search

Treponema pallidum subsp. pallidum can invade the central nervous system (CNS) at various stages of the disease (2, 11). CNS invasion by T. pallidum is determined by a positive syph- ilis serologic test, abnormal cerebrospinal fluid (CSF) cell count and protein level, and\\/or a positive CSF Venereal Dis- ease Research Laboratory (VDRL) test. Despite the high specificity of the CSF

A. V. VILLANUEVA; R. P. PODZORSKI; M. P. REYES

59

Biology of Treponema pallidum: Correlation of Functional Activities With Genome Sequence Data  

Microsoft Academic Search

Aspects of the biology of T. pallidum subsp. pallidum, the agent of syphilis, are examined in the context of a century of experimental studies and the recently determined genome sequence. T. pallidum and a group of closely related pathogenic spirochetes have evolved to become highly invasive, persistent pathogens with little toxigenic activity and an inability to survive outside the mammalian

Steven J. Norris; David L. Cox; George M. Weinstock

60

The genome of Treponema pallidum: new light on the agent of syphilis  

Microsoft Academic Search

Treponema pallidum subsp. pallidum, the causative agent of the sexually transmitted disease syphilis, is a fastidious, microaerophilic obligate parasite of humans. This bacterium is one of the few prominent infectious agents that has not been cultured continuously in vitro and consequently relatively little is known about its virulence mechanisms at the molecular level. T. pallidum therefore represented an attractive candidate

George M. Weinstock; John M. Hardham; Michael P. McLeod; Erica J. Sodergren; Steven J. Norris

1998-01-01

61

In vitro culture system to determine MICs and MBCs of antimicrobial agents against Treponema pallidum subsp. pallidum (Nichols strain).  

PubMed

A new procedure for determining the susceptibility of Treponema pallidum subsp. pallidum to antimicrobial agents was developed, utilizing a tissue culture system which promotes the in vitro multiplication of this organism. In the absence of antibiotics, T. pallidum (Nichols virulent strain) multiplied an average of 10-fold when incubated for 7 days in the presence of Sf1Ep cottontail rabbit epithelial cell cultures. Varied concentrations of penicillin G, tetracycline, erythromycin, and spectinomycin were added to triplicate cultures to determine their effects on treponemal multiplication, motility, and virulence. The MIC of each antibiotic was defined as the lowest concentration which prevented treponemal multiplication, whereas the MBC was defined as the lowest concentration which abrogated the ability of the cultured treponemes to multiply and cause lesions in rabbits. The in vitro culture technique provided highly reproducible MICs and (in parentheses) MBCs of each of the antibiotics tested: aqueous penicillin G, 0.0005 (0.0025) microgram/ml; tetracycline, 0.2 (0.5) microgram/ml; erythromycin, 0.005 (0.005) microgram/ml; and spectinomycin, 0.5 (0.5) microgram/ml. The significance of these results in light of the in vivo activities and the previous in vitro evaluations of these antibiotics is discussed. The T. pallidum in vitro cultivation system shows promise as a method for studying the interaction between T. pallidum and antimicrobial agents and for screening new antibiotics for syphilis therapy. PMID:2964810

Norris, S J; Edmondson, D G

1988-01-01

62

Global Challenge of Antibiotic-Resistant Treponema pallidum?  

PubMed Central

Syphilis is a multistage infectious disease that is usually transmitted through contact with active lesions of a sexual partner or from an infected pregnant woman to her fetus. Despite elimination efforts, syphilis remains endemic in many developing countries and has reemerged in several developed countries, including China, where a widespread epidemic recently occurred. In the absence of a vaccine, syphilis control is largely dependent upon identification of infected individuals and treatment of these individuals and their contacts with antibiotics. Although penicillin is still effective, clinically significant resistance to macrolides, a second-line alternative to penicillin, has emerged. Macrolide-resistant strains of Treponema pallidum are now prevalent in several developed countries. An understanding of the genetic basis of T. pallidum antibiotic resistance is essential to enable molecular surveillance. This review discusses the genetic basis of T. pallidum macrolide resistance and the potential of this spirochete to develop additional antibiotic resistance that could seriously compromise syphilis treatment and control. PMID:19805553

Stamm, Lola V.

2010-01-01

63

Effects of molecular oxygen, oxidation-reduction potential, and antioxidants upon in vitro replication of Treponema pallidum subsp. pallidum.  

PubMed Central

The effects of various concentrations of dithiothreitol, molecular oxygen, and several antioxidants upon the in vitro replication of Treponema pallidum were studied. The optimal dithiothreitol concentration was between 0.65 and 1.62 mM, and the optimum oxygen concentration was 3.0% +/- 0.5% in both the presence and absence of additional antioxidants. It was discovered that the reduced sulfhydryl concentration and the oxidation-reduction potential of the medium were stabilized after 5 days. The water-soluble antioxidants cobalt chloride, cocarboxylase, mannitol, and histidine were individually tested for their ability to increase treponemal growth in vitro. The optimum concentrations for these antioxidants were 21 nM, 4.3 nM, 0.55 mM, and 0.23 mM, respectively. When combined at these concentrations, the mixture of antioxidants stimulated the in vitro replication of T. pallidum. The number of treponemes in cultures with the antioxidants averaged a 59-fold increase, compared with a 43-fold increase in cultures lacking the antioxidants. It was further demonstrated that histidine and mannitol were the most critical components of this mixture. Catalase and superoxide dismutase were investigated for their ability to promote the growth and maintain viability of T. pallidum in tissue culture. The optimum concentrations for these enzymes were 10,000 U/liter and 25,000 U/liter, respectively. When these enzymes and the above antioxidants were combined and added to a chemically reduced modified Eagle medium, the treponemes increased an average of 70-fold, compared with an average of 35-fold in cultures lacking them. Furthermore, this medium, T. pallidum culture medium, supported the replication of T. pallidum at oxygen concentrations from 5 to 7% with little loss in yield or viability. The lipid-soluble antioxidants vitamin A and vitamin E acetate were also shown to enhance the in vitro growth of T. pallidum in this medium. PMID:2285317

Cox, D L; Riley, B; Chang, P; Sayahtaheri, S; Tassell, S; Hevelone, J

1990-01-01

64

The role of the ventral pallidum in psychiatric  

E-print Network

The role of the ventral pallidum in psychiatric disorders Largely garnered from studies are being substan- tiated, and the role of the VP in the human emotional repertoire and psychiatric. For example, single-photon emission computed tomography of Parkinson's Disease (PD) patients with pathologi

65

In vitro Cultivation and Purification of Treponema pallidum.  

National Technical Information Service (NTIS)

Studies utilizing both normal rabbit testes and ME 180 human tumor-derived monolayer cells have shown that T. pallidum (1) attaches to and enters the monolayer cells within 30 minutes, (2) attaches to cells by an active treponemal process, (3) persists in...

J. N. Miller

1975-01-01

66

Diagnosing Treponema pallidum in Secondary Syphilis by PCR and Immunohistochemistry  

Microsoft Academic Search

Epidemiological aspects of syphilis in Western countries have undergone a significant change with respect to the number of cases. Detection of Treponema pallidum is difficult, and the correct diagnosis of secondary syphilis can be critical. In this study, biopsy samples from skin lesions of 12 patients with secondary syphilis were used. Diagnosis of syphilis was based on clinical presentation, dark-field

Marc Buffet; Philippe A Grange; Philippe Gerhardt; Agnčs Carlotti; Vincent Calvez; Anne Bianchi; Nicolas Dupin

2007-01-01

67

Catabolism of glucose and fatty acids by virulent Treponema pallidum.  

PubMed Central

We describe a procedure which permits essentially full recovery of physiologically active Treponema pallidum from rabbit testicular extracts and greatly reduces contaminating tissue material. Such preparations were employed for investigations of the ability of T. pallidum to catabolize glucose and fatty acids. Radiorespirometric studies revealed that glucose and pyruvate, but not oleate or palmitate, could be degraded to CO2. The use of differentially labeled glucose, in conjunction with enzymatic analyses, indicated that glucose was catabolized by a combination of the Embden-Meyerhoff-Parnas and hexose monophosphate pathways. Pyruvate was degraded to CO2 from only the carboxyl position, suggesting the absence of a functioning Krebs cycle; this was substantiated by additional enzyme analyses and radiorespirometric experiments. Oleate and palmitate were incorporated but not catabolized by beta-oxidation. Glucose, although catabolized, was not incorporated. The potential significance of these findings is discussed. PMID:326678

Schiller, N L; Cox, C D

1977-01-01

68

THE DIRECT CULTIVATION OF TREPONEMA PALLIDUM PATHOGENIC FOR THE MONKEY  

PubMed Central

A method for the direct cultivation of Treponema pallidum from human syphilitic lesions, by the employment of a solid medium, has been described. By means of it, three of the four strains worked with were successfully cultivated. The several pure cultures agree in morphological and cultural characters, grow only in the presence of sterile tissue under anaerobic conditions, and do not produce putrefactive odors. The morphology is typical under optimum cultural conditions; it becomes atypical when the conditions are unfavorable. In cultures, Treponema pallidum multiplies by longitudinal division. The process is usually symmetrical but occasionally appears to be asymmetrical. Inoculation of the pure cultures into the skin of two species of lower monkeys was followed by the production of lesions resembling the primary syphilitic lesion occurring in man and those caused in the monkey by inoculation of spirochćtć-containing serum from human sources. During the course of the positive inoculation in the monkey, the blood develops the property of giving a positive Wassermann reaction. Thus the relation of Treponema pallidum to this reaction is supported, and the identity of the cultivated strains with the species found in human syphilitic lesions established. PMID:19867508

Noguchi, Hideyo

1912-01-01

69

T-Cell Responses to Treponema pallidum subsp. pallidum Antigens during the Course of Experimental Syphilis Infection  

Microsoft Academic Search

In this study we describe the development of the T-cell response to a panel of Treponema pallidum antigens over the course of syphilis infection in the rabbit and determine whether these antigens induce the expression of Th1 cytokines. It was determined that the membrane proteins TpN17 and TpN47, as well as the endoflagellar sheath protein TpN37, induce strong proliferation responses

THOMAS W. ARROLL; ARTURO CENTURION-LARA; SHEILA A. LUKEHART; WESLEY C. VAN VOORHIS

70

Two 16S-23S ribosomal DNA intergenic regions in different Treponema pallidum subspecies contain tRNA genes  

Microsoft Academic Search

The 16S-23S intergenic spacers of Treponema pallidum subspecies pallidum, Nichols strain, and Treponema pallidum subspecies pertenue, Gauthier strain, have been cloned, characterized and sequenced. Isoleucine and alanine tRNA genes have been identified within the 16S-23S intergenic regions on separate alleles of 293 and 303 bases, respectively. The two alleles are present in both T.p. pallidum and T.p. pertenue, and show

Arturo Centurion-Lara; Christa Castro; Wesley C. Van Voorhis; Sheila A. Lukehart

1996-01-01

71

Genome Analyses of T. pallidum and B. burgdorferi 387 Genome Analyses of Spirochetes: A Study of the  

E-print Network

of the Protein Structures, Functions and Metabolic Pathways in Treponema pallidum and Borrelia burgdorferi *For) and Treponema pallidum (Fraser et al., 1998), two pathogenic prokaryotic spirochetes, have generated a lotGenome Analyses of T. pallidum and B. burgdorferi 387 Genome Analyses of Spirochetes: A Study

Gerstein, Mark

72

A redescripton of Lyrosoma pallidum (Eschscholtz) and distributional range extension of Lyrosoma Mannerheim (Coleoptera, Agyrtidae)  

PubMed Central

Abstract A redescription with illustrations of the species Lyrosoma pallidum and a key to the Korean species of the family Agyrtidae are provided. New distributional data, including a range extension, of the two Lyrosoma Mannerheim species are presented. Lyrosoma pallidum (Eschscholtz) is recorded for the first time in Korea. PMID:24146551

Yoo, In-Seong; Sikes, Derek; Ahn, Kee-Jeong

2013-01-01

73

Plasmid DNA in Treponema pallidum (Nichols): Potential for Antibiotic Resistance by Syphilis Bacteria  

Microsoft Academic Search

A plasmid DNA structure (approximate molecular weight = 7.5 × 106) was identified in the human pathogen Treponema pallidum (Nichols). The inability to isolate this plasmid from rabbit host tissue and the total lack of DNA homology of the plasmid with rabbit DNA has confirmed its Treponema pallidum origin. The observation documents a newly recognized and potentially significant genetic capability

Michael V. Norgard; James N. Miller

1981-01-01

74

TprC/D (Tp0117/131), a Trimeric, Pore-Forming Rare Outer Membrane Protein of Treponema pallidum, Has a Bipartite Domain Structure  

PubMed Central

Identification of Treponema pallidum rare outer membrane proteins (OMPs) has been a longstanding objective of syphilis researchers. We recently developed a consensus computational framework that employs a battery of cellular localization and topological prediction tools to generate ranked clusters of candidate rare OMPs (D. L. Cox et al., Infect. Immun. 78:5178–5194, 2010). TP0117/TP0131 (TprC/D), a member of the T. pallidum repeat (Tpr) family, was a highly ranked candidate. Circular dichroism, heat modifiability by SDS-PAGE, Triton X-114 phase partitioning, and liposome incorporation confirmed that full-length, recombinant TprC (TprCFl) forms a ?-barrel capable of integrating into lipid bilayers. Moreover, TprCFl increased efflux of terbium-dipicolinic acid complex from large unilamellar vesicles and migrated as a trimer by blue-native PAGE. We found that in T. pallidum, TprC is heat modifiable, trimeric, expressed in low abundance, and, based on proteinase K accessibility and opsonophagocytosis assays, surface exposed. From these collective data, we conclude that TprC is a bona fide rare OMP as well as a functional ortholog of Escherichia coli OmpF. We also discovered that TprC has a bipartite architecture consisting of a soluble N-terminal portion (TprCN), presumably periplasmic and bound directly or indirectly to peptidoglycan, and a C-terminal ?-barrel (TprCC). Syphilitic rabbits generate antibodies exclusively against TprCC, while secondary syphilis patients fail to mount a detectable antibody response against either domain. The syphilis spirochete appears to have resolved a fundamental dilemma arising from its extracellular lifestyle, namely, how to enhance OM permeability without increasing its vulnerability to the antibody-mediated defenses of its natural human host. PMID:22389487

Anand, Arvind; Luthra, Amit; Dunham-Ems, Star; Caimano, Melissa J.; Karanian, Carson; LeDoyt, Morgan; Cruz, Adriana R.; Salazar, Juan C.

2012-01-01

75

TprC/D (Tp0117/131), a trimeric, pore-forming rare outer membrane protein of Treponema pallidum, has a bipartite domain structure.  

PubMed

Identification of Treponema pallidum rare outer membrane proteins (OMPs) has been a longstanding objective of syphilis researchers. We recently developed a consensus computational framework that employs a battery of cellular localization and topological prediction tools to generate ranked clusters of candidate rare OMPs (D. L. Cox et al., Infect. Immun. 78:5178-5194, 2010). TP0117/TP0131 (TprC/D), a member of the T. pallidum repeat (Tpr) family, was a highly ranked candidate. Circular dichroism, heat modifiability by SDS-PAGE, Triton X-114 phase partitioning, and liposome incorporation confirmed that full-length, recombinant TprC (TprC(Fl)) forms a ?-barrel capable of integrating into lipid bilayers. Moreover, TprC(Fl) increased efflux of terbium-dipicolinic acid complex from large unilamellar vesicles and migrated as a trimer by blue-native PAGE. We found that in T. pallidum, TprC is heat modifiable, trimeric, expressed in low abundance, and, based on proteinase K accessibility and opsonophagocytosis assays, surface exposed. From these collective data, we conclude that TprC is a bona fide rare OMP as well as a functional ortholog of Escherichia coli OmpF. We also discovered that TprC has a bipartite architecture consisting of a soluble N-terminal portion (TprC(N)), presumably periplasmic and bound directly or indirectly to peptidoglycan, and a C-terminal ?-barrel (TprC(C)). Syphilitic rabbits generate antibodies exclusively against TprC(C), while secondary syphilis patients fail to mount a detectable antibody response against either domain. The syphilis spirochete appears to have resolved a fundamental dilemma arising from its extracellular lifestyle, namely, how to enhance OM permeability without increasing its vulnerability to the antibody-mediated defenses of its natural human host. PMID:22389487

Anand, Arvind; Luthra, Amit; Dunham-Ems, Star; Caimano, Melissa J; Karanian, Carson; LeDoyt, Morgan; Cruz, Adriana R; Salazar, Juan C; Radolf, Justin D

2012-05-01

76

Ultrastructural features of malignant syphilis and demonstration of Treponema pallidum.  

PubMed

This paper reports a case of malignant syphilis (man, 39 years old) in whom ultrastructural investigations of a typical nodule revealed an extremely low amount of bacteria with the characteristics of Treponema pallidum in poorly differentiated cells of the dermal infiltrate with plasma cells, stimulated lymphocytes, and neutrophils as predominating cell types. Most of the microorganisms bore signs of disintegration. Vascular changes and exocytosis were only demonstrable by light microscopy in a second nodule. Together with the high production rate of immunoglobulins and an excessive inflammatory reaction, these findings point to an aberrant biologic reaction pattern of those patients who develop malignant syphilis. Unfortunately, further investigations concerning a possible impairment of cellular immunity as supposed in the literature, had not been possible in the present case. PMID:6343268

Bahmer, F A; Anton-Lamprecht, L

1983-04-01

77

Isolation of the outer membranes from Treponema pallidum and Treponema vincentii.  

PubMed Central

The outer membranes from Treponema pallidum subsp. pallidum and Treponema vincentii were isolated by a novel method. Purified outer membranes from T. pallidum and T. vincentii following sucrose gradient centrifugation banded at 7 and 31% (wt/wt) sucrose, respectively. Freeze fracture electron microscopy of purified membrane vesicles from T. pallidum and T. vincentii revealed an extremely low density of protein particles; the particle density of T. pallidum was approximately six times less than that of T. vincentii. The great majority of T. vincentii lipopolysaccharide was found in the outer membrane preparation. The T. vincentii outer membrane also contained proteins of 55 and 65 kDa. 125I-penicillin V labeling demonstrated that t. pallidum penicillin-binding proteins were found exclusively with the protoplasmic cylinders and were not detectable with purified outer membrane material, indicating the absence of inner membrane contamination. Isolated T. pallidum outer membrane was devoid of the 19-kDa 4D protein and the normally abundant 47-kDa lipoprotein known to be associated with the cytoplasmic membrane; only trace amounts of the periplasmic endoflagella were detected. Proteins associated with the T. pallidum outer membrane were identified by one- and two-dimensional electrophoretic analysis using gold staining and immunoblotting. Small amounts of strongly antigenic 17- and 45-kDa proteins were detected and shown to correspond to previously identified lipoproteins which are found principally with the cytoplasmic membrane. Less antigenic proteins of 65, 31 (acidic pI), 31 (basic pI), and 28 kDa were identified. Compared with whole-organism preparations, the 65- and the more basic 31-kDa proteins were found to be highly enriched in the outer membrane preparation, indicating that they may represent the T. pallidum rare outer membrane proteins. Reconstitution of solubilized T. pallidum outer membrane into lipid bilayer membranes revealed porin activity with two estimated channel diameters of 0.35 and 0.68 nm based on the measured single-channel conductances in 1 M KCl of 0.40 and 0.76 nS, respectively. Images PMID:7928971

Blanco, D R; Reimann, K; Skare, J; Champion, C I; Foley, D; Exner, M M; Hancock, R E; Miller, J N; Lovett, M A

1994-01-01

78

Identification of the Treponema pallidum subsp. pallidum TP0092 (RpoE) Regulon and Its Implications for Pathogen Persistence in the Host and Syphilis Pathogenesis  

PubMed Central

Bacteria often respond to harmful environmental stimuli with the induction of extracytoplasmic function (ECF) sigma (?) factors that in turn direct RNA polymerase to transcribe specific groups of response genes (or regulons) to minimize cellular damage and favor adaptation to the changed extracellular milieu. In Treponema pallidum subsp. pallidum, the agent of syphilis, the TP0092 gene is predicted to code for the pathogen's only annotated ECF ? factor, homologous to RpoE, known in Escherichia coli to control a key transduction pathway for maintenance of envelope homeostasis in response to external stress and cell growth. Here we have shown that TP0092 is highly transcribed during experimental syphilis. Furthermore, TP0092 transcription levels significantly increase as infection progresses toward immune clearance of the pathogen, suggesting a role for TP0092 in helping T. pallidum respond to harmful stimuli in the host environment. To investigate this hypothesis, we determined the TP0092 regulon at two different time points during infection using chromatin immunoprecipitation followed by high-throughput sequencing. A total of 22 chromosomal regions, all containing putative TP0092-binding sites and corresponding to as many T. pallidum genes, were identified. Noteworthy among them are the genes encoding desulfoferrodoxin and thioredoxin, involved in detoxification of reactive oxygen species (ROS). Because T. pallidum does not possess other enzymes for ROS detoxification, such as superoxide dismutase, catalase, or glutathione peroxidase, our results suggest that the TP0092 regulon is important in protecting the syphilis spirochete from damage caused by ROS produced at the site of infection during the inflammatory response. PMID:23243302

Denisenko, Oleg; Tompa, Martin; Centurion-Lara, Arturo

2013-01-01

79

MyD88 Deficiency Markedly Worsens Tissue Inflammation and Bacterial Clearance in Mice Infected with Treponema pallidum, the Agent of Syphilis  

PubMed Central

Research on syphilis, a sexually transmitted infection caused by the non-cultivatable spirochete Treponema pallidum, has been hampered by the lack of an inbred animal model. We hypothesized that Toll-like receptor (TLR)-dependent responses are essential for clearance of T. pallidum and, consequently, compared infection in wild-type (WT) mice and animals lacking MyD88, the adaptor molecule required for signaling by most TLRs. MyD88-deficient mice had significantly higher pathogen burdens and more extensive inflammation than control animals. Whereas tissue infiltrates in WT mice consisted of mixed mononuclear and plasma cells, infiltrates in MyD88-deficient animals were predominantly neutrophilic. Although both WT and MyD88-deficient mice produced antibodies that promoted uptake of treponemes by WT macrophages, MyD88-deficient macrophages were deficient in opsonophagocytosis of treponemes. Our results demonstrate that TLR-mediated responses are major contributors to the resistance of mice to syphilitic disease and that MyD88 signaling and FcR-mediated opsonophagocytosis are linked to the macrophage-mediated clearance of treponemes. PMID:23940747

Silver, Adam C.; Dunne, Dana W.; Zeiss, Caroline J.; Bockenstedt, Linda K.; Radolf, Justin D.; Salazar, Juan C.; Fikrig, Erol

2013-01-01

80

[Generalized infantile neuroaxonal dystrophies with pigmentation and lipophanerosis of the pallidum in concordant twins (author's transl)].  

PubMed

Monozygotic male twins died at the age of 6 1/2 and 7 1/2 years respectively after a progressive course of mental deterioration, hypotonia, spasticity, optic atrophy and seizures that had commenced at the age of 2 years. Both patients showed generalized neuroaxonal dystrophy (NAD), marked by numerous spheroids, iron-positive pigment and lipophanerosis of the pallidum. NAD can be classified as a generalized form without pigmentation of the pallidum (infantile type of Seitelberger), a juvenile type of Rozdilsky, a generalized form with pigmentation (cases described here), and localized forms (infantile, late infantile, juvenile = classic Hallervorden-Spatz disease, adult types). PMID:183173

Peiffer, J; Brunner, N; Landolt, R F; Müller, G; Schlote, W

1976-08-01

81

MOLECULAR CHARACTERIZATION OF RECEPTOR BINDING PROTEINS AND IMMUNOGENS OF VIRULENT TREPONEMA PALLIDUM  

Microsoft Academic Search

During the clinical course of syphilis, a complex interrelationship exists between virulent Treponema pallidum and the parasitized host. Infection can persist in the presence of a significant immune response, and manifestations of actively developing disease are well documented (1, 2). Although controversies still remain concei'ning the relative contributions of cellular and humoral immunity to eradication of the disease, it is

JOEL B. BASEMAN; EDWARD C. HAYES

82

The immune response to infection with Treponema pallidum, the stealth pathogen  

Microsoft Academic Search

Cutaneous immunobiology and spirochetal molecular biology have allowed investigators to propose a conceptual framework for the development of both the innate and adaptive immune response to Treponema pallidum infection. While some clinical manifestations can be attributed to humoral responses, most can be attributed to a combination of local innate and adaptive cellular immunity.

Juan C. Salazar; Karsten R. O. Hazlett; Justin D. Radolf

2002-01-01

83

TP0326, a Treponema pallidum ?-Barrel Assembly Machinery A (BamA) Ortholog and Rare Outer Membrane Protein  

PubMed Central

SUMMARY Definitive identification of Treponema pallidum (Tp) rare outer membrane proteins (OMPs) has long eluded researchers. TP0326, the sole protein in Tp with sequence homology to a Gram-negative OMP, belongs to the BamA family of proteins essential for OM biogenesis. Structural modeling predicted that five polypeptide transport-associated (POTRA) domains comprise the N-terminus of TP0326, while the C-terminus forms an 18-stranded amphipathic ?-barrel. Circular dichroism, heat-modifiability by SDS-PAGE, Triton X-114 phase partitioning and liposome incorporation supported these topological predictions and confirmed that the ?-barrel is responsible for the native protein's amphiphilicity. Expression analyses revealed that native TP0326 is expressed at low abundance, while a protease-surface accessibility assay confirmed surface exposure. Size-exclusion chromatography and blue native polyacrylamide gel electrophoresis revealed a modular Bam complex in Tp considerably larger than that of E. coli. Non-orthologous ancillary factors and self-association of TP0326 via its ?-barrel may both contribute to the Bam complex. Tp-infected rabbits mount a vigorous antibody response to both POTRA and ?-barrel portions of TP0326, whereas humans with secondary syphilis respond predominantly to POTRA. The syphilis spirochete appears to have devised a stratagem for harnessing the Bam pathway while satisfying its need to limit surface antigenicity. PMID:21488980

Desrosiers, Daniel C.; Anand, Arvind; Luthra, Amit; Dunham-Ems, Star M; LeDoyt, Morgan; Cummings, Michael A. D.; Eshghi, Azad; Cameron, Caroline E.; Cruz, Adriana R.; Salazar, Juan C.; Caimano, Melissa J.; Radolf, Justin D.

2011-01-01

84

Treponema pallidum Infection in the Wild Baboons of East Africa: Distribution and Genetic Characterization of the Strains Responsible  

PubMed Central

It has been known for decades that wild baboons are naturally infected with Treponema pallidum, the bacterium that causes the diseases syphilis (subsp. pallidum), yaws (subsp. pertenue), and bejel (subsp. endemicum) in humans. Recently, a form of T. pallidum infection associated with severe genital lesions has been described in wild baboons at Lake Manyara National Park in Tanzania. In this study, we investigated ten additional sites in Tanzania and Kenya using a combination of macroscopic observation and serology, in order to determine whether the infection was present in each area. In addition, we obtained genetic sequence data from six polymorphic regions using T. pallidum strains collected from baboons at two different Tanzanian sites. We report that lesions consistent with T. pallidum infection were present at four of the five Tanzanian sites examined, and serology was used to confirm treponemal infection at three of these. By contrast, no signs of treponemal infection were observed at the six Kenyan sites, and serology indicated T. pallidum was present at only one of them. A survey of sexually mature baboons at Lake Manyara National Park in 2006 carried out as part of this study indicated that roughly ten percent displayed T. pallidum-associated lesions severe enough to cause major structural damage to the genitalia. Finally, we found that T. pallidum strains from Lake Manyara National Park and Serengeti National Park were genetically distinct, and a phylogeny suggested that baboon strains may have diverged prior to the clade containing human strains. We conclude that T. pallidum infection associated with genital lesions appears to be common in the wild baboons of the regions studied in Tanzania. Further study is needed to elucidate the infection's transmission mode, its associated morbidity and mortality, and the relationship between baboon and human strains. PMID:23284649

Harper, Kristin N.; Fyumagwa, Robert D.; Hoare, Richard; Wambura, Philemon N.; Coppenhaver, Dorian H.; Sapolsky, Robert M.; Alberts, Susan C.; Tung, Jenny; Rogers, Jeffrey; Kilewo, Morris; Batamuzi, Emmanuel K.; Leendertz, Fabian H.; Armelagos, George J.; Knauf, Sascha

2012-01-01

85

Two Mutations associated with Macrolide Resistance in Treponema pallidum: Increasing Prevalence and Correlation with Molecular Strain Type in Seattle, Washington  

PubMed Central

Background Although azithromycin promised to be a safe and effective single dose oral treatment for early syphilis, azithromycin treatment failure has been documented and is associated with mutations in the 23S rDNA of corresponding Treponema pallidum strains. The prevalence of strains harboring these mutations varies throughout the US and the world. We examined T. pallidum strains circulating in Seattle, Washington, from 2001–2010 to determine the prevalence of two mutations associated with macrolide resistance, and to determine whether these mutations were associated with certain T. pallidum strain types. Methods Subjects were enrolled in a separate ongoing study of cerebrospinal fluid (CSF) abnormalities in patients with syphilis. T. pallidum DNA purified from blood and T. pallidum strains isolated from blood or CSF were analyzed for two 23S rDNA mutations and for the molecular targets used in an enhanced molecular stain typing system. Results Nine molecular strain types of T. pallidum were identified in Seattle from 2001–2010. Both macrolide resistance mutations were identified in Seattle strains, and the prevalence of resistant T. pallidum exceeded 80% in 2005 and increased through 2010. Resistance mutations were associated with discrete molecular strain types of T. pallidum. Conclusions Macrolide resistant T. pallidum strains are highly prevalent in Seattle, and each mutation is associated with discrete strain types. Macrolides should not be considered for treatment of syphilis in regions where prevalence of the mutations is high. Combining the resistance mutations with molecular strain typing permits a finer analysis of the epidemiology of syphilis in a community. PMID:23191949

Grimes, Matthew; Sahi, Sharon K.; Godornes, B. Charmie; Tantalo, Lauren C.; Roberts, Neal; Bostick, David; Marra, Christina M.; Lukehart, Sheila A.

2013-01-01

86

Evaluation of the recombinant protein TpF1 of Treponema pallidum for serodiagnosis of syphilis.  

PubMed

Syphilis is a chronic infection caused by Treponema pallidum subsp. pallidum, and diagnosis with sensitive and specific methods is a challenging process that is important for its prevention and treatment. In the present study, we established a recombinant protein TpF1-based indirect immunoglobulin G (IgG) enzyme-linked immunosorbent assay (ELISA) and a Western blot assay for human and rabbit sera. The 20-kDa recombinant protein TpF1 was detected by Western blotting performed with sera from rabbits immunized with recombinant TpF1 and infected with the T. pallidum Nichols strain and T. pallidum clinical isolates but was not detected by Western blotting with sera from uninfected rabbits. The sensitivity of the recombinant protein was determined by screening sera from individuals with primary, secondary, latent, and congenital syphilis (n = 82). The specificity of the recombinant protein was determined by screening sera from uninfected controls (n = 30) and individuals with potentially cross-reactive infections, including Lyme disease (n = 30) and leptospirosis (n = 5). The sensitivities of TpF1-based ELISAs were 93.3%, 100%, 100%, and 100% for primary, secondary, latent, and congenital syphilis, respectively, and the specificities were all 100% for sera from uninfected controls and individuals with potentially cross-reactive infections. In Western blot assays, the sensitivities and specificities of TpF1 for human sera were all 100%. The reactivities of TpF1 with syphilitic sera were proportional to the titers of the T. pallidum particle agglutination (TPPA) assay. These data indicate that the recombinant protein TpF1 is a highly immunogenic protein in human and rabbit infections and a promising marker for the screening of syphilis. PMID:23945159

Jiang, Chuanhao; Zhao, Feijun; Xiao, Jinhong; Zeng, Tiebing; Yu, Jian; Ma, Xiaohua; Wu, Haiying; Wu, Yimou

2013-10-01

87

Evaluation of the Recombinant Protein TpF1 of Treponema pallidum for Serodiagnosis of Syphilis  

PubMed Central

Syphilis is a chronic infection caused by Treponema pallidum subsp. pallidum, and diagnosis with sensitive and specific methods is a challenging process that is important for its prevention and treatment. In the present study, we established a recombinant protein TpF1-based indirect immunoglobulin G (IgG) enzyme-linked immunosorbent assay (ELISA) and a Western blot assay for human and rabbit sera. The 20-kDa recombinant protein TpF1 was detected by Western blotting performed with sera from rabbits immunized with recombinant TpF1 and infected with the T. pallidum Nichols strain and T. pallidum clinical isolates but was not detected by Western blotting with sera from uninfected rabbits. The sensitivity of the recombinant protein was determined by screening sera from individuals with primary, secondary, latent, and congenital syphilis (n = 82). The specificity of the recombinant protein was determined by screening sera from uninfected controls (n = 30) and individuals with potentially cross-reactive infections, including Lyme disease (n = 30) and leptospirosis (n = 5). The sensitivities of TpF1-based ELISAs were 93.3%, 100%, 100%, and 100% for primary, secondary, latent, and congenital syphilis, respectively, and the specificities were all 100% for sera from uninfected controls and individuals with potentially cross-reactive infections. In Western blot assays, the sensitivities and specificities of TpF1 for human sera were all 100%. The reactivities of TpF1 with syphilitic sera were proportional to the titers of the T. pallidum particle agglutination (TPPA) assay. These data indicate that the recombinant protein TpF1 is a highly immunogenic protein in human and rabbit infections and a promising marker for the screening of syphilis. PMID:23945159

Jiang, Chuanhao; Zhao, Feijun; Xiao, Jinhong; Zeng, Tiebing; Yu, Jian; Ma, Xiaohua; Wu, Haiying

2013-01-01

88

Bifunctional Role of the Treponema pallidum Extracellular Matrix Binding Adhesin Tp0751 ?  

PubMed Central

Treponema pallidum, the causative agent of syphilis, is a highly invasive pathogenic spirochete capable of attaching to host cells, invading the tissue barrier, and undergoing rapid widespread dissemination via the circulatory system. The T. pallidum adhesin Tp0751 was previously shown to bind laminin, the most abundant component of the basement membrane, suggesting a role for this adhesin in host tissue colonization and bacterial dissemination. We hypothesized that similar to that of other invasive pathogens, the interaction of T. pallidum with host coagulation proteins, such as fibrinogen, may also be crucial for dissemination via the circulatory system. To test this prediction, we used enzyme-linked immunosorbent assay (ELISA) methodology to demonstrate specific binding of soluble recombinant Tp0751 to human fibrinogen. Click-chemistry-based palmitoylation profiling of heterologously expressed Tp0751 confirmed the presence of a lipid attachment site within this adhesin. Analysis of the Tp0751 primary sequence revealed the presence of a C-terminal putative HEXXH metalloprotease motif, and in vitro degradation assays confirmed that recombinant Tp0751 purified from both insect and Escherichia coli expression systems degrades human fibrinogen and laminin. The proteolytic activity of Tp0751 was abolished by the presence of the metalloprotease inhibitor 1,10-phenanthroline. Further, inductively coupled plasma-mass spectrometry showed that Tp0751 binds zinc and calcium. Collectively, these results indicate that Tp0751 is a zinc-dependent, membrane-associated protease that exhibits metalloprotease-like characteristics. However, site-directed mutagenesis of the HEXXH motif to HQXXH did not abolish the proteolytic activity of Tp0751, indicating that further mutagenesis studies are required to elucidate the critical active site residues associated with this protein. This study represents the first published description of a T. pallidum protease capable of degrading host components and thus provides novel insight into the mechanism of T. pallidum dissemination. PMID:21149586

Houston, Simon; Hof, Rebecca; Francescutti, Teresa; Hawkes, Aaron; Boulanger, Martin J.; Cameron, Caroline E.

2011-01-01

89

Characterization of a manganese-dependent regulatory protein, TroR, from Treponema pallidum  

PubMed Central

Genome sequence analysis of Treponema pallidum, the causative agent of syphilis, suggests that this bacterium has a limited iron requirement with few, if any, proteins that require iron. Instead, T. pallidum may use manganese-dependent enzymes for metabolic pathways. This strategy apparently alleviates the necessity of T. pallidum to acquire iron from the host, thus overcoming iron limitation, which is a primary host defense. Interestingly, a putative metal-dependent regulatory protein, TroR, which has homology with the diphtheria toxin regulatory protein, DtxR, from Corynebacterium diphtheriae was identified from T. pallidum. We describe here the characterization of TroR, a regulatory protein. Mobility-shift DNA binding and DNase I footprint assays indicated that purified TroR bound to a 22-nt region of dyad symmetry that overlaps the ?10 region of the promoter of the tro operon, which contains the genes for a putative metal transport system, the glycolytic enzyme phosphoglycerate mutase, and TroR. Unlike other metal-dependent regulatory proteins like diphtheria toxin regulatory protein and the ferric ion uptake regulator, Fur, which can be activated by divalent metals such as Fe2+, Mn2+, Co2+, Ni2+, and Zn2+, TroR is activated only by Mn2+. The TroR-Mn2+ complex binds its target sequence and blocks transcription of the troPO/lacZ fusion, suggesting that TroR acts as a metal-dependent repressor in vivo. In addition, TroR exists as a dimer in both its inactive (metal free) and active states as indicated by chemical crosslinking experiments. Based on these data, we propose that TroR represents a unique regulatory system for controlling gene expression in T. pallidum in response to Mn2+. PMID:10485921

Posey, James E.; Hardham, John M.; Norris, Steven J.; Gherardini, Frank C.

1999-01-01

90

The application of a protein fraction derived from Treponema Pallidum (reiter strain) as an antigen in the serodiagnosis of syphilis  

Microsoft Academic Search

Summary  A comparison was made between the complement fixation test using a protein fraction derived fromTreponema pallidum (Reiter strain) as an antigen and theTreponema pallidum immobilization test. As appears from the results obtained with 116 syphilitic and 137 presumably non-syphilitic sera the\\u000a complement fixation test with protein antigen showed an excellent sensitivity together with a satisfactory specificity.

J. H. De Bruijn

1957-01-01

91

NUTRITION OF CELLULAR SLIME MOLDS II. Growth of Polysphondylium pallidum in Axenic Culture  

PubMed Central

Hohl, Hans-Rudolf (University of Wisconsin, Madison) and Kenneth B. Raper. Nutrition of cellular slime molds. II. Growth of Polysphondylium pallidum in axenic culture. J. Bacteriol. 85:199–206. 1963.—Several strains of Polysphondylium pallidum were grown on a liquid soluble medium in axenic culture. The medium contained embryo extract, serum albumin, Tryptose, dextrose, vitamins, and salts. The final cell yield was about 6–11 × 106 cells/ml, depending on the strain. The generation time was usually about 5 to 6 hr. The myxamoebae were grown for over 125 generations on this soluble complex medium without decrease in growth vigor or loss of their capacity to form normal fructifications when removed to an appropriate surface (e.g., agar). Thus the whole life cycle of this species was completed in the absence of any bacteria or bacterial products. Other species of the Dictyosteleaceae grew less well or failed to grow in the liquid medium described. PMID:13961229

Hohl, Hans-Rudolf; Raper, Kenneth B.

1963-01-01

92

Cryo-electron tomography elucidates the molecular architecture of Treponema pallidum, the syphilis spirochete.  

PubMed

Cryo-electron tomography (CET) was used to examine the native cellular organization of Treponema pallidum, the syphilis spirochete. T. pallidum cells appeared to form flat waves, did not contain an outer coat and, except for bulges over the basal bodies and widening in the vicinity of flagellar filaments, displayed a uniform periplasmic space. Although the outer membrane (OM) generally was smooth in contour, OM extrusions and blebs frequently were observed, highlighting the structure's fluidity and lack of attachment to underlying periplasmic constituents. Cytoplasmic filaments converged from their attachment points opposite the basal bodies to form arrays that ran roughly parallel to the flagellar filaments along the inner surface of the cytoplasmic membrane (CM). Motile treponemes stably attached to rabbit epithelial cells predominantly via their tips. CET revealed that T. pallidum cell ends have a complex morphology and assume at least four distinct morphotypes. Images of dividing treponemes and organisms shedding cell envelope-derived blebs provided evidence for the spirochete's complex membrane biology. In the regions without flagellar filaments, peptidoglycan (PG) was visualized as a thin layer that divided the periplasmic space into zones of higher and lower electron densities adjacent to the CM and OM, respectively. Flagellar filaments were observed overlying the PG layer, while image modeling placed the PG-basal body contact site in the vicinity of the stator-P-collar junction. Bioinformatics and homology modeling indicated that the MotB proteins of T. pallidum, Treponema denticola, and Borrelia burgdorferi have membrane topologies and PG binding sites highly similar to those of their well-characterized Escherichia coli and Helicobacter pylori orthologs. Collectively, our results help to clarify fundamental differences in cell envelope ultrastructure between spirochetes and gram-negative bacteria. They also confirm that PG stabilizes the flagellar motor and enable us to propose that in most spirochetes motility results from rotation of the flagellar filaments against the PG. PMID:19820083

Izard, Jacques; Renken, Christian; Hsieh, Chyong-Ere; Desrosiers, Daniel C; Dunham-Ems, Star; La Vake, Carson; Gebhardt, Linda L; Limberger, Ronald J; Cox, David L; Marko, Michael; Radolf, Justin D

2009-12-01

93

Purification, antitumor and antioxidant activities in vitro of polysaccharides from the brown seaweed Sargassum pallidum  

Microsoft Academic Search

Supercritical CO2 extraction, ultrasonic-aid extraction and membrane separation technology were applied to prepare Sargassum pallidum polysaccharides (SP). Three main fractions, SP-1, SP-2 and SP-3, were obtained by membranes of 1.0×10?4mm pore size and normal molecular-weight cut-off of 50kDa. The resulting three preparations were further purified by DEAE Cellulose-52 chromatography to afford seven polysaccharide fractions. Furthermore, the antitumor and antioxidant activities,

Hong Ye; Keqi Wang; Chunhong Zhou; Jun Liu; Xiaoxiong Zeng

2008-01-01

94

Cryo-Electron Tomography Elucidates the Molecular Architecture of Treponema pallidum, the Syphilis Spirochete? †  

PubMed Central

Cryo-electron tomography (CET) was used to examine the native cellular organization of Treponema pallidum, the syphilis spirochete. T. pallidum cells appeared to form flat waves, did not contain an outer coat and, except for bulges over the basal bodies and widening in the vicinity of flagellar filaments, displayed a uniform periplasmic space. Although the outer membrane (OM) generally was smooth in contour, OM extrusions and blebs frequently were observed, highlighting the structure's fluidity and lack of attachment to underlying periplasmic constituents. Cytoplasmic filaments converged from their attachment points opposite the basal bodies to form arrays that ran roughly parallel to the flagellar filaments along the inner surface of the cytoplasmic membrane (CM). Motile treponemes stably attached to rabbit epithelial cells predominantly via their tips. CET revealed that T. pallidum cell ends have a complex morphology and assume at least four distinct morphotypes. Images of dividing treponemes and organisms shedding cell envelope-derived blebs provided evidence for the spirochete's complex membrane biology. In the regions without flagellar filaments, peptidoglycan (PG) was visualized as a thin layer that divided the periplasmic space into zones of higher and lower electron densities adjacent to the CM and OM, respectively. Flagellar filaments were observed overlying the PG layer, while image modeling placed the PG-basal body contact site in the vicinity of the stator-P-collar junction. Bioinformatics and homology modeling indicated that the MotB proteins of T. pallidum, Treponema denticola, and Borrelia burgdorferi have membrane topologies and PG binding sites highly similar to those of their well-characterized Escherichia coli and Helicobacter pylori orthologs. Collectively, our results help to clarify fundamental differences in cell envelope ultrastructure between spirochetes and gram-negative bacteria. They also confirm that PG stabilizes the flagellar motor and enable us to propose that in most spirochetes motility results from rotation of the flagellar filaments against the PG. PMID:19820083

Izard, Jacques; Renken, Christian; Hsieh, Chyong-Ere; Desrosiers, Daniel C.; Dunham-Ems, Star; La Vake, Carson; Gebhardt, Linda L.; Limberger, Ronald J.; Cox, David L.; Marko, Michael; Radolf, Justin D.

2009-01-01

95

Ultrastructure of treponema pallidum Nichols following lysis by physical and chemical methods  

Microsoft Academic Search

1.Normal cells of the Nichols non-pathogenic strain of Treponema pallidum were disrupted by one or a combination of physical and chemical procedures. Cells and cellular fragments were then observed in the electron microscope by means of three techniques: negative staining, thin sectioning, and freeze-etching. Changes in typical morphology were noted and individual organelles were isolated for detailed structural analysis.2.The cell

Sally Jackson; S. H. Black

1971-01-01

96

Fibronectin mediates Treponema pallidum cytadherence through recognition of fibronectin cell-binding domain  

Microsoft Academic Search

Numerous studies (1-5) suggest that an early event in the pathogenesis of syphilis is Treponema pallidum adherence to host cells (cytadherence), mediated by tip-like structures on virulent spirochetes. Biochemical-molecular studies have identified three treponemal outer envelope proteins as putative ligands respon- sible for surface parasitism (3, 6). These proteins are also highly immunogenic, as demonstrated by radioimmunoassays using sera from

D. DENISE THOMAS; JOEL B. BASEMAN; JOHN F. ALDERETE

1985-01-01

97

Characterization of Outer Membranes Isolated from Treponema pallidum, the Syphilis Spirochete  

Microsoft Academic Search

Previous freeze-fracture electron microscopy (EM) studies have shown that the outer membrane (OM) of Treponema pallidumcontains sparse transmembrane proteins. One strategy for molecular characterization of these rare OM proteins involves isolation ofT. pallidumOMs. Here we describe a simple and extremely gentle method for OM isolation based upon isopycnic sucrose density gradient ultracentrifugation of treponemes following plasmolysis in 20% sucrose. Evidence

JUSTIN D. RADOLF; ESTHER J. ROBINSON; KENNETH W. BOURELL; DARRIN R. AKINS; STEPHEN F. PORCELLA; LINDA M. WEIGEL; JEFFREY D. JONES; ANDMICHAEL V. NORGARD

1995-01-01

98

Macrolide treatment failure in a case of secondary syphilis: a novel A2059G mutation in the 23S rRNA gene of Treponema pallidum subsp. pallidum.  

PubMed

We report an occurrence of treatment failure after oral spiramycin therapy in a man with secondary syphilis and a reported penicillin and tetracycline allergy. Molecular detection revealed treponemal DNA in the blood of the patient and sequencing of the 23S rDNA identified an A to G transition at the gene position corresponding to position 2059 in the Escherichia coli 23S rRNA gene. The occurrence of this novel 23S rDNA mutation was examined among 7 rabbit-propagated syphilitic strains of Treponema pallidum and among 22 syphilis patient isolates from the Czech Republic. The prevalence of A2058G and A2059G mutations among clinical specimens was 18.2 and 18.2 %, respectively. PMID:19429763

Matejková, Petra; Flasarová, Magdalena; Zákoucká, Hana; Borek, Milan; Kremenová, Sona; Arenberger, Petr; Woznicová, Vladana; Weinstock, George M; Smajs, David

2009-06-01

99

Effect of heavy metals (Cu, Pb, and As) on the ultrastructure of Sargassum pallidum in Daya Bay, China.  

PubMed

Concentrations of Cu, Pb, and As were determined in seawater, surface sediment, Sargassum pallidum collected from the Daya Bay, China. The influence of metal contamination on the marine alga was investigated at chemical and ultrastructural level. Mean concentrations of Cu (19.44 mg kg(-1)) and Pb (33.99 mg kg(-1)) were found to be high in sediment, whereas concentration of As (122.29 mg kg(-1)) in S. pallidum was higher than that in water and sediment. The ultrastructure of S. pallidum cells was anomalous and aberrant. Energy-dispersive x-ray spectroscopic analysis revealed that the nanometal particles in the form of comparatively high-electron density substance diffused in the cell structures constituted by Cu, Pb, As, etc. There is a remarkable similarity or correspondence in the anomalous elements between the geochemistry and the botanic cell, and the heavy metals have potential hazardous effect on the ocean ecology system in Daya Bay. PMID:23982302

Miao, Li; Yan, Wen; Zhong, Lifeng; Xu, Weihai

2014-01-01

100

A longitudinal evaluation of Treponema pallidum PCR testing in early syphilis  

PubMed Central

Background Syphilis is a growing public health problem among men who have sex with men (MSM) globally. Rapid and accurate detection of syphilis is vital to ensure patients and their contacts receive timely treatment and reduce ongoing transmission. Methods We evaluated a PCR assay for the diagnosis of Treponema pallidum using swabs of suspected early syphilis lesions in longitudinally assessed MSM. Results We tested 260 MSM for T pallidum by PCR on 288 occasions: 77 (26.7%) had early syphilis that was serologically confirmed at baseline or within six weeks, and 211 (73.3%) remained seronegative for syphilis. Of 55 men with primary syphilis, 49 were PCR positive, giving a sensitivity of 89.1% (95% CI: 77.8%-95.9%) and a specificity of 99.1% (95% CI: 96.5%-99.9%). Of 22 men with secondary syphilis, 11 were PCR positive, giving a sensitivity of 50% (95% CI: 28.2%-71.8%) and a specificity of 100% (95% CI: 66.4%-71.8%). Of the 77 syphilis cases, 43 (56%) were HIV positive and the sensitivity and specificity of the PCR test did not vary by HIV status. The PCR test was able to detect up to five (10%) primary infections that were initially seronegative, including one HIV positive man with delayed seroconversion to syphilis (72 to 140 days) and one HIV positive man who did not seroconvert to syphilis over 14 months follow-up. Both men had been treated for syphilis within a week of the PCR test. Conclusions T pallidum PCR is a potentially powerful tool for the early diagnosis of primary syphilis, particularly where a serological response has yet to develop. PMID:23241398

2012-01-01

101

[Biological activity of lipids and photosynthetic pigments of Sargassum pallidum C. Agardh].  

PubMed

The biological activity of lipids and photosynthetic pigments of the kelp Sargassum pallidum (Turner) C. Agardh has been studied. Free fatty acids and their esters demonstrated considerable antimicrobial activity against bacteria (Staphylococcus aureus[ital] and Escherichia coli), yeast-like fungi (Candida albicans), and opportunistic pathogenic (Aspergilius niger) and phytopathogenic (Fusarium oxysporum, and Septoria glycines) fungi. Glyceroglycolipids and neutral lipids demonstrated moderate activity. Fucoxanthin and chlorophylls weakly suppressed the growth of microorganisms. None of the studied substances demonstrated activity against Ehrlich's carcinoma. It was shown that the season of weed harvesting affected both antimicrobial and hemolytic activities of different lipids due to changes in their fatty acid composition. PMID:25272757

Gerasimenko, N I; Martyias, E A; Logvinov, S V; Busarova, N G

2014-01-01

102

Evaluation of an immunochromatographic point-of-care test for the simultaneous detection of nontreponemal and treponemal antibodies in patients with syphilis.  

PubMed

We described the evaluation of the Syphilis Screening & Confirm Assay for the simultaneous detection of nontreponemal and treponemal antibodies. A total of 248 samples were evaluated. The sensitivity of the tests was 98.8%, 99.5% and 98.9%, while specificity was 94.7%, 88.9% and 93.2%, respectively, as compared with the rapid plasma reagin, Treponema pallidum hemagglutination assay, and fluorescent treponemal antibody absorption tests. PMID:25013972

Castro, Rita; Lopes, Ângela; da Luz Martins Pereira, Filomena

2014-08-01

103

Structural, Bioinformatic, and In Vivo Analyses of Two Treponema pallidum Lipoproteins Reveal a Unique TRAP Transporter  

SciTech Connect

Treponema pallidum, the bacterial agent of syphilis, is predicted to encode one tripartite ATP-independent periplasmic transporter (TRAP-T). TRAP-Ts typically employ a periplasmic substrate-binding protein (SBP) to deliver the cognate ligand to the transmembrane symporter. Herein, we demonstrate that the genes encoding the putative TRAP-T components from T. pallidum, tp0957 (the SBP), and tp0958 (the symporter), are in an operon with an uncharacterized third gene, tp0956. We determined the crystal structure of recombinant Tp0956; the protein is trimeric and perforated by a pore. Part of Tp0956 forms an assembly similar to those of 'tetratricopeptide repeat' (TPR) motifs. The crystal structure of recombinant Tp0957 was also determined; like the SBPs of other TRAP-Ts, there are two lobes separated by a cleft. In these other SBPs, the cleft binds a negatively charged ligand. However, the cleft of Tp0957 has a strikingly hydrophobic chemical composition, indicating that its ligand may be substantially different and likely hydrophobic. Analytical ultracentrifugation of the recombinant versions of Tp0956 and Tp0957 established that these proteins associate avidly. This unprecedented interaction was confirmed for the native molecules using in vivo cross-linking experiments. Finally, bioinformatic analyses suggested that this transporter exemplifies a new subfamily of TPATs (TPR-protein-associated TRAP-Ts) that require the action of a TPR-containing accessory protein for the periplasmic transport of a potentially hydrophobic ligand(s).

Deka, Ranjit K.; Brautigam, Chad A.; Goldberg, Martin; Schuck, Peter; Tomchick, Diana R.; Norgard, Michael V. (NIH); (UTSMC)

2012-05-25

104

A synthetic lymph node containing inactivated Treponema pallidum cells elicits strong, antigen-specific humoral and cellular immune responses in mice.  

PubMed

The goal of this study was to investigate the use of a synthetic lymph node (SLN) for delivery of Treponema pallidum (Tp) antigens. Immune responses of C57BL/6 mice were analyzed at 4, 8, and 12 weeks after SLN implantation. Group 1 mice received SLN with no antigen; Group 2, SLN with formalin-inactivated Tp (f-Tp); and Group 3, SLN with f-Tp plus a CpG oligodeoxynucleotide. When tested by ELISA, sera from Group 2 and Group 3 mice showed stronger IgG antibody reactivity than sera from Group 1 mice to sonicates of f-Tp or untreated Tp, but not to sonicate of normal rabbit testicular extract at all times. The IgG1 level was higher than IgG2c level for Group 2 mice at all times and for Group 3 mice at 4 and 8 weeks. IgG1 and IgG2c levels were nearly equivalent for Group 3 mice at 12 weeks. Immunoblotting showed that IgG from Group 2 and Group 3 mice recognized several Tp proteins at all times. Supernatants of splenocytes from Group 2 and Group 3 mice contained significantly more IFN? than those from Group 1 mice after stimulation with f-Tp at all times. A significant level of IL-4 was not detected in any supernatants. These data show that strong humoral and cellular immune responses to Tp can be elicited via a SLN. PMID:24106125

Stamm, Lola V; Drapp, Rebecca L

2014-02-01

105

Designer Receptors Show Role for Ventral Pallidum Input to Ventral Tegmental Area in Cocaine Seeking  

PubMed Central

Ventral pallidum (VP) is centrally positioned within mesocorticolimbic reward circuits, and its dense projection to ventral tegmental area (VTA) regulates neuronal activity there. However, VP is a heterogeneous structure, and how this complexity affects its role within wider reward circuits is unclear. Here we demonstrate that projections to VTA from rostral (RVP), but not caudal VP (CVP) are robustly Fos-activated during cue-induced reinstatement of cocaine seeking—a rat model of relapse in addiction. Moreover, designer receptor-mediated transient inactivation of RVP neurons, their terminals in VTA, or functional connectivity between RVP and VTA dopamine neurons blocks the ability of drug-associated cues (but not a cocaine prime) to reinstate cocaine seeking. In contrast, CVP neuronal inhibition instead blocked cocaine-primed, but not cue-induced reinstatement. This novel double dissociation in VP sub-regional roles in drug seeking is likely important for understanding mesocorticolimbic circuits underlying reward seeking and addiction. PMID:24584054

Mahler, Stephen V.; Vazey, Elena M.; Beckley, Jacob T; Keistler, Colby R.; McGlinchey, Ellen M.; Kaufling, Jennifer; Wilson, Steven P.; Deisseroth, Karl; Woodward, John J.; Aston-Jones, Gary

2014-01-01

106

Treponema pallidum 3Phosphoglycerate Mutase Is a Heat-Labile Enzyme That May Limit the Maximum Growth Temperature for the Spirochete  

Microsoft Academic Search

In the causative agent of syphilis, Treponema pallidum, the gene encoding 3-phosphoglycerate mutase, gpm, is part of a six-gene operon (tro operon) that is regulated by the Mn-dependent repressor TroR. Since substrate-level phosphorylation via the Embden-Meyerhof pathway is the principal way to generate ATP in T. pallidum and Gpm is a key enzyme in this pathway, Mn could exert a

STEPHANE BENOIT; JAMES E. POSEY; MATTHEW R. CHENOWETH; FRANK C. GHERARDINI

2001-01-01

107

Comparison of the Serodia Treponema pallidum Particle Agglutination, Captia Syphilis-G, and SpiroTek Reagin II Tests with Standard Test Techniques for Diagnosis of Syphilis  

Microsoft Academic Search

We compared the microhemagglutination assay for Treponema pallidum (MHA-TP), a treponemal test, with two other treponemal tests, the Serodia Treponema pallidum particle agglutination (TP-PA) assay and the Captia Syphilis-G enzyme immunoassay, using 390 clinical serum samples. We also compared two nontrepo- nemal tests, the rapid plasma Reagin (RPR) card test and the SpiroTek Reagin II test. Agreements of the MHA-TP

VICTORIA POPE; MARTHA B. FEARS; WILLIAM E. MORRILL; ARNOLD CASTRO; SUSAN E. KIKKERT

2001-01-01

108

Tromp1, a Putative Rare Outer Membrane Protein, Is Anchored by an Uncleaved Signal Sequence to the Treponema pallidum Cytoplasmic Membrane  

Microsoft Academic Search

Treponema pallidum rare outer membrane protein 1 (Tromp1) has extensive sequence homology with sub- strate-binding proteins of ATP-binding cassette transporters. Because such proteins typically are periplasmic or cytoplasmic membrane associated, experiments were conducted to clarify Tromp1's physicochemical prop- erties and cellular location in T. pallidum. Comparison of the sodium dodecyl sulfate-polyacrylamide gel electrophoresis mobilities of (i) native Tromp1 and Tromp1

DARRIN R. AKINS; ESTHER ROBINSON; DMITRIY SHEVCHENKO; CHRISTOPHER ELKINS; DAVID L. COX; JUSTIN D. RADOLF

1997-01-01

109

Identification and transcriptional analysis of a Treponema pallidum operon encoding a putative ABC transport system, an iron-activated repressor protein homolog, and a glycolytic pathway enzyme homolog  

Microsoft Academic Search

We have characterized a 5.2-kilobase (kb) putative transport related operon (tro) locus of Treponema pallidum subsp. pallidum (Nichols strain) (Tp) encoding six proteins: TroA, TroB, TroC, TroD, TroR and Phosphoglycerate mutase (Pgm). Four of these gene products (TroA–TroD) are homologous to members of the ATP-Binding Cassette (ABC) superfamily of bacterial transport proteins. TroA (previously identified as Tromp1) has significant sequence

John M. Hardham; Lola V. Stamm; Stephen F. Porcella; Jonathan G. Frye; Natalie Y. Barnes; Jerrilyn K. Howell; Stacey L. Mueller; Justin D. Radolf; George M. Weinstock; Steven J. Norris

1997-01-01

110

Genetic characterization and partial sequence determination of a Treponema pallidum operon expressing two immunogenic membrane proteins in Escherichia coli.  

PubMed Central

A detailed physical and genetic map of a previously cloned 5.5-kilobase segment of Treponema pallidum DNA is described. This segment expressed two proteins that are cell membrane associated in Escherichia coli. The structural genes of these treponemal membrane proteins, tmpA and tmpB, are coordinately expressed, and transcription in E. coli can start from at least two different treponemal promoters. The tmpA and tmpB proteins are the products of in vivo proteolytic cleavage from precursor proteins which are 2 and 4 kilodaltons larger, respectively, than the mature proteins. Because the sizes of the corresponding proteins produced in T. pallidum were identical to those of the mature membrane proteins in E. coli, we concluded that a similar proteolytic processing takes place in both E. coli and T. pallidum. Although tmpA and tmpB were controlled by the same transcription signals, tmpB was expressed to a higher extent than tmpA, and only the tmpB product could be overproduced by placing the left lambda promoter in front of the structural genes. The nucleotide sequence of the T. pallidum tmpA gene was established. This is the first T. pallidum gene sequenced. Codon usage and the nature of transcriptional and translational signals are discussed. The deduced amino acid sequence indicated the presence of a sequence that was characteristic for a signal peptide. This sequence information allowed the construction of hybrid genes coding for proteins having beta-galactosidase enzyme activity as well as TmpA epitopes. The enzyme-linked antigen was expressed at a high level in E. coli when transcriptional and translational signals from coliphage lambda were used. In this case the protein produced was a sandwich protein consisting of 21 amino acids of the lambda cro protein, 204 amino acids of the T. pallidum TmpA protein, and 1,020 amino acids of the E. coli lambda-galactosidase. The potential use of this enzyme-linked antigen for the serodiagnosis of syphilis is discussed. Images PMID:3922944

Hansen, E B; Pedersen, P E; Schouls, L M; Severin, E; van Embden, J D

1985-01-01

111

Identification and sequence analysis of Treponema pallidum tprJ, a member of a polymorphic multigene family  

Microsoft Academic Search

TnphoA mutagenesis was used to identify genes encoding exported proteins in a genomic DNA library of Treponema pallidum, the syphilis agent. The nucleotide sequence of an open reading frame (tprJ) that encodes a 755-amino acid protein with a predicted molecular mass of 81.1 kDa was determined. The deduced amino acid sequence of TprJ has homology to the major surface protein

Lola V. Stamm; Shermalyn R. Greene; Heather L. Bergen; John M. Hardham; Natalie Y. Barnes

1998-01-01

112

Diagnosis of congenital syphilis from placental examination: Comparison of histopathology, steiner stain, and polymerase chain reaction for Treponema pallidum DNA  

Microsoft Academic Search

Congenital syphilis is often a presumptive diagnosis (based on serologies), because confirmation requires identification of Treponema pallidum in fetal\\/neonatal tissues or in the placenta. Placental histological features associated with congenital syphilis include the triad of enlarged hypercellular villi, proliferative fetal vascular changes, and acute or chronic villitis. The authors blindly evaluated 49 formalin-fixed, paraffin-embedded placentas (38 with positive maternal syphilis

David R Genest; Sung R Choi-Hong; James E Tate; Faisal Qureshi; Suzanne M Jacques; Christopher Crum

1996-01-01

113

Physicochemical Evidence that Treponema pallidum TroA Is a Zinc-Containing Metalloprotein That Lacks Porin-Like Structure  

Microsoft Academic Search

Although TroA (Tromp1) was initially reported to be a Treponema pallidum outer membrane protein with porin-like properties, subsequent studies have suggested that it actually is a periplasmic substrate-binding protein involved in the transport of metals across the treponemal cytoplasmic membrane. Here we conducted additional physicochemical studies to address the divergent viewpoints concerning this protein. Triton X-114 phase partitioning of recombinant

RANJIT K. DEKA; YONG-HWAN LEE; KAYLA E. HAGMAN; DMITRIY SHEVCHENKO; CLIFFORD A. LINGWOOD; CHARLES A. HASEMANN; MICHAEL V. NORGARD; JUSTIN D. RADOLF

1999-01-01

114

Monoclonal antibodies and immobilized antibodies  

Microsoft Academic Search

Antibodies in both their free and immobilized state have been the object of considerable industrial and academic interest.\\u000a A variety of methods are used for preparing and immobilizing antibodies. Applications for monoclonal antibodies include the\\u000a preparation of therapeutics, diagnostics, and in affinity fractionation. Recent US patents on monoclonal and immobilized antibodies\\u000a and scientific literature on monoclonal antibodies are surveyed. A

Robert J. Linhardt; C. W. Abell; R. M. Denney; B. W. Altrock; R. Auerbach; S. D. Bernal; R. E. Canfield; P. H. Ehrlich; W. R. Moyle; T. S. Chan; T. W. Chang; N. T. Chang; J. A. Cidlowski; M. D. Viceps; R. J. Cote; D. M. Morrissey; A. N. Houghton; E. J. Beattie; H. F. Oettgen; L. J. Old; C. M. Croce; R. S. Cubicciotti; A. E. Karu; R. M. Krauss; J. S. Cullor; A. Deutsch; H. Brandwein; H. Platt; D. M. Hunter; A. Dubitsky; S. M. Durham; F. A. Dolbeare; J. W. Gray; G. R. Dreesman; C. E. Kendall; J. C. Egrie; A. R. Frackelton; H. N. Eisen; A. H. Ross; S. Gay; G. Geirnaert; J. E. Geltosky; E. H. Goldberg; E. Goldwasser; C. Kavinsky; T. L. Weiss; H. G. Gratzner; B. Hampar; M. Zweig; S. D. Showalter; H. H. Handley; M. C. Glassy; Y. Hagiwara; H. Hagiwara; C. M. Huang; S. N. Cohen; J. V. Hughes; E. M. Scolnick; J. E. Tomassini; R. Jefferis; J. Steensgaard; H. S. Kaplan; N. N. H. Teng; K. S. Earn; R. F. Calvo; L. Kass; J. R. Kettman; M. V. Norgard; M. B. Khazaeli; W. H. Beierwaltes; B. G. England; P. C. Kung; G. Goldstein; L. Lanier; J. Phillips; N. L. Warner; J. W. Larrick; A. R. Raubitschek; K. E. Truitt; H. Lazarus; J. F. Schwaber; J. Lewicki; C. Lewis; J. V. Olander; W. R. Tolbert; E. L. Milford; C. B. Carpenter; J. M. Paradysz; D. F. Mosher; J. L. Mulshine; J. D. Minna; K. A. Murray; D. M. Neville; R. J. Youle; M. Nicolson; I. Pastan; M. C. Willingham; D. J. Fitzgerald; A. Pucci; A. M. Smithyman; M. B. Slade; P. W. French; G. Wijffels; C. S. Pukel; K. O. Lloyd; L. R. Travassos; W. G. Dippold; R. P. Reckel; J. L. Harris; R. Wellerson; S. M. Shaw; P. M. Kaplan; E. L. Reinherz; S. F. Schlossman; S. C. Mener; J. Sakamoto; C. C. Cordon; E. Friedman; C. L. Finstad; W. E. Enker; M. R. Melamed; J. F. Oettgen; P. J. Scannon; L. E. Spitler; H. M. Lee; R. T. Kawahata; R. P. Mischak; J. Schlom; D. Colcher; M. Nuti; P. H. Hand; F. Austin; G. D. Shockman; D. E. Jackson; W. Wong; Z. Steplewski; H. Koprowski; M. Herlyn; M. Strand; I. S. Trowbridge; D. L. Urdal; C. J. March; S. K. Dower; J. R. Wands; V. R. Zurawski; C. A. White; R. Dulbecco; W. R. Allen; E. C. Arnold; M. Flasher; H. H. Freedman; T. D. Heath; P. Shek; D. Papahadjopoulos; M. Ikeda; S. Sakamoto; K. Suzuki; M. Kuboyama; Y. Harada; A. Kawashiri; E. Takahashi; H. S. Lee; S. Margel; R. C. Nowinski; A. S. Hoffman; J. W. Peterson; K. B. Platt; D. E. Reed; F. X. Real; M. J. Mattes; P. O. Livingston; A. Rembaum; R. C. K. Yen; R. Rosenstein; B. Schneider

1987-01-01

115

Repellency of cassia bark, eucalyptus, and star anise oils and their major constituents to Leptotrombidium pallidum (Acari: Trombiculidae).  

PubMed

Leptotrombidium pallidum (Nagoya, Miyagawa, Mitamura & Tamiya) is a primary vector of Orientia tsutsugamushi (Hyashi), the causative agent of scrub typhus. An assessment is made of the repellency to L. pallidum larvae (chiggers) of cassia bark, eucalyptus, and star anise oils and major constituents (E)-cinnamaldehyde, 1,8-cineole, and (E)-anethole of the corresponding oils. Results were compared with those of conventional repellents DEET (N,N-diethyl-3-methylbenzamide), IR3535 [(ethyl 3-[acetyl(butyl)amino]propanoate)], and permethrin. Based on the median repellent concentration (RC50) values, (E)-cinnamaldehyde, (E)-anethole, cassia bark oil, and star anise oil (RC50, 0.95-1.52 mg/cm2) exhibited significantly more potent repellency than DEET (3.85 mg/cm2). (E)-cinnamaldehyde, (E)-anethole, cassiabark oil, 1,8-cineole, and star anise oil were approximately 43, 16, 11, 8, and 4 times more effective than IR3535 (CC5, 6.51%) as judged by the median climbing distance-disturbing concentration (CC50) values. The median residual duration time of repellency (RT50) was significantly more pronounced in DEET (RT50, 323 min) than in all essential oils and constituents (108-167 min). In the light of global efforts to reduce the level of highly toxic synthetic repellents, the three essential oils and their major constituents described merit further study as potential biorepellents for the control of L. pallidum populations. PMID:23802452

Shin, E-Hyun; Song, Bong Gu; Lee, Il Hee; Park, Mi Yeoun; Ahn, Young-Joon; Chang, Kyu-Sik

2013-05-01

116

Biophysical and Bioinformatic Analyses Implicate the Treponema pallidum Tp34 Lipoprotein (Tp0971) in Transition Metal Homeostasis  

PubMed Central

Metal ion homeostasis is a critical function of many integral and peripheral membrane proteins. The genome of the etiologic agent of syphilis, Treponema pallidum, is compact and devoid of many metabolic enzyme genes. Nevertheless, it harbors genes coding for homologs of several enzymes that typically require either iron or zinc. The product of the tp0971 gene of T. pallidum, designated Tp34, is a periplasmic lipoprotein that is thought to be tethered to the inner membrane of this organism. Previous work on a water-soluble (nonacylated) recombinant version of Tp34 established that this protein binds to Zn2+, which, like other transition metal ions, stabilizes the dimeric form of the protein. In this study, we employed analytical ultracentrifugation to establish that four transition metal ions (Ni2+, Co2+, Cu2+, and Zn2+) readily induce the dimerization of Tp34; Cu2+ (50% effective concentration [EC50] = 1.7 ?M) and Zn2+ (EC50 = 6.2 ?M) were the most efficacious of these ions. Mutations of the crystallographically identified metal-binding residues hindered the ability of Tp34 to dimerize. X-ray crystallography performed on crystals of Tp34 that had been incubated with metal ions indicated that the binding site could accommodate the metals examined. The findings presented herein, coupled with bioinformatic analyses of related proteins, point to Tp34's likely role in metal ion homeostasis in T. pallidum. PMID:23042995

Brautigam, Chad A.; Deka, Ranjit K.; Ouyang, Zhiming; Machius, Mischa; Knutsen, Gregory; Tomchick, Diana R.

2012-01-01

117

Monoclonal Antibodies.  

ERIC Educational Resources Information Center

Monoclonal antibodies have provided an exciting addition to the "armory" of the molecular biologist and immunologist. This article discusses briefly the concept of, techniques available for, production of, and possible uses of monoclonal antibodies. (Author)

Killington, R. A.; Powell, K. L.

1984-01-01

118

BIOCHEMICAL CHANGES DURING GROWTH AND ENCYSTMENT OF THE CELLULAR SLIME MOLD POLYSPHONDYLIUM PALLIDUM  

PubMed Central

The growth of the cellular slime mold, Polysphondylium pallidum, was studied on a semidefined medium in shaken suspension. When the medium contained large quantities of particulate material, growth was more rapid and the cellular size and protein content were smaller than when growth occurred on a medium containing less particulate material. The cellular levels of DNA, RNA, and protein; of lysosomal enzymes (acid phosphatase, acid proteinase); and of peroxisomal enzymes (catalase) were assayed during growth and the subsequent stationary phase that led eventually to encystment. Only DNA remained at a constant cellular level. Encystment of exponentially growing cells could also be initiated by washing them and introducing them into a soluble peptone medium. The rate of encystment was proportional to the osmolarity of this medium. The encystment process was followed with respect to the cellular levels of DNA, RNA, protein, carbohydrates, acid phosphatase, acid ?-N-Ac-glucosaminidase, and catalase. The most dramatic change occurred in the cellular cellulose content, which increased by at least an order of magnitude by the time encystment was morphologically complete. It was concluded that the encystment of this slime mold in suspension exhibits a number of biochemical similarities to the development of this and other cellular slime molds on a surface. PMID:4795859

Githens, S.; Karnovsky, M. L.

1973-01-01

119

Mitochondrial tRNA 5'-editing in Dictyostelium discoideum and Polysphondylium pallidum.  

PubMed

Mitochondrial tRNA (mt-tRNA) 5'-editing was first described more than 20 years ago; however, the first candidates for 5'-editing enzymes were only recently identified in a eukaryotic microbe (protist), the slime mold Dictyostelium discoideum. In this organism, eight of 18 mt-tRNAs are predicted to be edited based on the presence of genomically encoded mismatched nucleotides in their aminoacyl-acceptor stem sequences. Here, we demonstrate that mt-tRNA 5'-editing occurs at all predicted sites in D. discoideum as evidenced by changes in the sequences of isolated mt-tRNAs compared with the expected sequences encoded by the mitochondrial genome. We also identify two previously unpredicted editing events in which G-U base pairs are edited in the absence of any other genomically encoded mismatches. A comparison of 5'-editing in D. discoideum with 5'-editing in another slime mold, Polysphondylium pallidum, suggests organism-specific idiosyncrasies in the treatment of U-G/G-U pairs. In vitro activities of putative D. discoideum editing enzymes are consistent with the observed editing reactions and suggest an overall lack of tRNA substrate specificity exhibited by the repair component of the editing enzyme. Although the presence of terminal mismatches in mt-tRNA sequences is highly predictive of the occurrence of mt-tRNA 5'-editing, the variability in treatment of U-G/G-U base pairs observed here indicates that direct experimental evidence of 5'-editing must be obtained to understand the complete spectrum of mt-tRNA editing events in any species. PMID:24737330

Abad, Maria G; Long, Yicheng; Kinchen, R Dimitri; Schindel, Elinor T; Gray, Michael W; Jackman, Jane E

2014-05-30

120

Differential roles of ventral pallidum subregions during cocaine self-administration behaviors  

PubMed Central

The ventral pallidum (VP) is necessary for drug-seeking behavior. VP contains ventromedial (VPvm) and dorsolateral (VPdl) subregions which receive projections from the nucleus accumbens shell and core, respectively. To date, no study has investigated the behavioral functions of the VPdl and VPvm subregions. To address this issue, we investigated whether changes in firing rate (FR) differed between VP subregions during four events: approaching toward, responding on, or retreating away from a cocaine-reinforced operandum, and a cocaine-associated cue. Baseline FR and waveform characteristics did not differ between subregions. VPdl neurons exhibited a greater change in FR compared to VPvm neurons during approaches toward, as well as responses on, the cocaine-reinforced operandum. VPdl neurons were more likely to exhibit a similar change in FR (direction and magnitude) during approach and response than VPvm neurons. In contrast, VPvm firing patterns were heterogeneous, changing FRs during approach or response alone, or both. VP neurons did not discriminate cued behaviors from uncued behaviors. No differences were found between subregions during the retreat and no VP neurons exhibited patterned changes in FR in response to the cocaine-associated cue. The stronger, sustained FR changes of VPdl neurons during approach and response may implicate VPdl in the processing of drug-seeking and drug-taking behavior via projections to subthalamic nucleus and substantia nigra pars reticulata. In contrast, heterogeneous firing patterns of VPvm neurons may implicate VPvm in facilitating mesocortical structures with information related to the sequence of behaviors predicting cocaine self-infusions via projections to mediodorsal thalamus and ventral tegmental area. PMID:22806483

Root, David H.; Ma, Sisi; Barker, David J.; Megehee, Laura; Striano, Brendan M.; Ralston, Carla M.; Fabbricatore, Anthony T.; West, Mark O.

2012-01-01

121

ANTIBODY FORMATION  

PubMed Central

The suppression of antibody formation by passively administered antibody is influenced by the dose and nature of the antigen, type of immunization procedure, ratio of antibody to antigen, species origin and characteristics of the antiserum used, as well as the species selected for immunization. In guinea pigs, diphtheria antitoxin formation can be effectively suppressed by an intravenous injection of excess homologous or heterologous antitoxin as long as 5 days after toxoid immunization and after delayed-type hypersensitivity to toxoid has developed. Following the period of antibody suppression which lasts 2 to 7 weeks, serum antibody can usually be demonstrated. It is proposed that this delayed immunization results from dissociation of antigen, since diphtheritic paralysis and death can be produced in guinea pigs and rabbits by the intravenous injection of toxin-antitoxin precipitates formed in antitoxin excess. This syndrome is prevented by injection of excess horse antitoxin 1 hour after injection of the toxin-antitoxin complexes. PMID:13779027

Uhr, Jonathan W.; Baumann, Joyce B.

1961-01-01

122

NMDA-induced lesions of the nucleus accumbens or the ventral pallidum increase the rewarding efficacy of food to deprived rats  

Microsoft Academic Search

The role of the nucleus accumbens (NAC) and ventral pallidum (VP) in food reward modulation was investigated using Heyman's [24] curve fitting approach in food deprived rats. All rats were maintained at 80% normal body weight, and trained to lever press for food reinforcement. Each rat was tested daily with a series of four variable-interval (VI) reinforcement schedules (80, 40,

Patricia I. Johnson; Mary Ann Parente; James R. Stellar

1996-01-01

123

The role of the ventral pallidum GABAergic system in conditioned taste aversion: Effects of microinjections of a GABA A receptor antagonist on taste palatability of a conditioned stimulus  

Microsoft Academic Search

When subjects receive a taste stimulus (conditioned stimulus, CS) that is paired with malaise, they acquire conditioned taste aversion (CTA). It is thought that the taste CS changes from appetitive to aversive after acquisition of CTA. Previous studies have suggested that the ventral pallidum (VP) is involved in the hedonics of taste stimuli, therefore the present study investigated whether the

Tadashi Inui; Tsuyoshi Shimura; Takashi Yamamoto

2007-01-01

124

Pharmacokinetic properties of arsenic species after oral administration of Sargassum pallidum extract in rats using an HPLC-HG-AFS method.  

PubMed

Sargassum pallidum is one of the Traditional Chinese Medicine widely used for phlegm elimination and detumescence. Arsenic is present in high concentration in seaweed belonging to the genus Sargassum. Therefore, the consumption of S. pallidum is a route of exposure to arsenic. Since the toxicity of arsenic is highly dependent on its chemical speciation, the determination of total arsenic is not adequate to assess the risks. Here, a high performance liquid chromatography-hydride generation-atomic fluorescence spectrometry (HPLC-HG-AFS) was developed for determination of the common arsenic species including arsenite [As(III)], dimethylarsinate (DMA), methylarsonate (MMA) and arsenate [As(V)] simultaneously. This method was applied to study the pharmacokinetic profile of these arsenic species in rats after oral administration of S. pallidum extract at different doses. The described assay was validated for limit of quantification, linearity, intra-day and inter-day precisions, accuracy, extraction recovery and stability according to the FDA validation guidelines. As(III) or MMA was not detected in any samples collected at all time points using the present HPLC-HG-AFS method. As(V) and DMA in the S. pallidum could be readily absorbed and eliminated in rats. A trend of dose-dependence was shown for DMA and As(V) in the drug concentration-time profiles. This study would be helpful for the apprehension of the action mechanism and clinical application of medicinal seaweeds. PMID:24763266

Cao, Yan; Duan, Jinao; Guo, Jianming; Li, Weixia; Tao, Weiwei

2014-08-01

125

Estimation of the genome sizes of the chigger mites Leptotrombidium pallidum and Leptotrombidium scutellare based on quantitative PCR and k-mer analysis  

PubMed Central

Background Leptotrombidium pallidum and Leptotrombidium scutellare are the major vector mites for Orientia tsutsugamushi, the causative agent of scrub typhus. Before these organisms can be subjected to whole-genome sequencing, it is necessary to estimate their genome sizes to obtain basic information for establishing the strategies that should be used for genome sequencing and assembly. Method The genome sizes of L. pallidum and L. scutellare were estimated by a method based on quantitative real-time PCR. In addition, a k-mer analysis of the whole-genome sequences obtained through Illumina sequencing was conducted to verify the mutual compatibility and reliability of the results. Results The genome sizes estimated using qPCR were 191?±?7 Mb for L. pallidum and 262?±?13 Mb for L. scutellare. The k-mer analysis-based genome lengths were estimated to be 175 Mb for L. pallidum and 286 Mb for L. scutellare. The estimates from these two independent methods were mutually complementary and within a similar range to those of other Acariform mites. Conclusions The estimation method based on qPCR appears to be a useful alternative when the standard methods, such as flow cytometry, are impractical. The relatively small estimated genome sizes should facilitate whole-genome analysis, which could contribute to our understanding of Arachnida genome evolution and provide key information for scrub typhus prevention and mite vector competence. PMID:24947244

2014-01-01

126

New Tests for Syphilis: Rational Design of a PCR Method for Detection of Treponema pallidum in Clinical Specimens Using Unique Regions of the DNA Polymerase I Gene  

Microsoft Academic Search

A sensitive and specific PCR method to detect Treponema pallidum in clinical specimens was developed. PCR primers were designed based on two unique features of the DNA polymerase I gene (polA). The first distinctive characteristic is that the region codes for a high cysteine content and has low homology with similar regions of DNA polymerase I gene from known microorganisms.

HSI LIU; BERTA RODES; C.-Y. CHEN; BRET STEINER

127

New Tests for Syphilis: Rational Design of a PCR Method for Detection of Treponema pallidum in Clinical Specimens Using Unique Regions of the DNA Polymerase I Gene  

Microsoft Academic Search

A sensitive and specific PCR method to detect Treponema pallidum in clinical specimens was developed. PCR primers were designed based on two unique features of the DNA polymerase I gene (polA). The first distinctive characteristic is that the region codes for a high cysteine content and has low homology with similar regions of DNA polymerase I gene from known microorganisms.

HSI LIU; BERTA RODES; C.-Y. Chen; BRET STEINER

2001-01-01

128

Assessment of the Kinetics of Treponema pallidum Dissemination into Blood and Tissues in Experimental Syphilis by Real-Time Quantitative PCR  

Microsoft Academic Search

Little is known about the size and kinetics of treponemal burdens in blood and tissues during acquired or experimental syphilitic infection. We used real-time quantitative PCR to measure Treponema pallidum DNA levels in rabbits infected intratesticularly with the prototype Nichols strain. At the outset, we performed a series of in vitro blood spiking experiments to determine the effect of blood

Juan C. Salazar; Asha Rathi; Nelson L. Michael; Justin D. Radolf; Linda L. Jagodzinski

2007-01-01

129

Development of a System for Expressing Heterologous Genes in the Oral Spirochete Treponema denticola and Its Use in Expression of the Treponema pallidum flaA Gene  

Microsoft Academic Search

Spirochetes have unique morphology and motility. Their periplasmic flagella, located between the outer membrane and the cytoplasmic membrane, play an important role in cellular morphology and motility (5, 15). The Treponema genus con- tains several important pathogens, and many of these patho- genic spirochetes cannot be cultured in vitro. One of the most important spirochete pathogens is Treponema pallidum, the

BO CHI; SARITA CHAUHAN; HOWARD KURAMITSU

1999-01-01

130

Treponema pallidum 3-phosphoglycerate mutase is a heat-labile enzyme that may limit the maximum growth temperature for the spirochete.  

PubMed

In the causative agent of syphilis, Treponema pallidum, the gene encoding 3-phosphoglycerate mutase, gpm, is part of a six-gene operon (tro operon) that is regulated by the Mn-dependent repressor TroR. Since substrate-level phosphorylation via the Embden-Meyerhof pathway is the principal way to generate ATP in T. pallidum and Gpm is a key enzyme in this pathway, Mn could exert a regulatory effect on central metabolism in this bacterium. To study this, T. pallidum gpm was cloned, Gpm was purified from Escherichia coli, and antiserum against the recombinant protein was raised. Immunoblots indicated that Gpm was expressed in freshly extracted infective T. pallidum. Enzyme assays indicated that Gpm did not require Mn(2+) while 2,3-diphosphoglycerate (DPG) was required for maximum activity. Consistent with these observations, Mn did not copurify with Gpm. The purified Gpm was stable for more than 4 h at 25 degrees C, retained only 50% activity after incubation for 20 min at 34 degrees C or 10 min at 37 degrees C, and was completely inactive after 10 min at 42 degrees C. The temperature effect was attenuated when 1 mM DPG was added to the assay mixture. The recombinant Gpm from pSLB2 complemented E. coli strain PL225 (gpm) and restored growth on minimal glucose medium in a temperature-dependent manner. Increasing the temperature of cultures of E. coli PL225 harboring pSLB2 from 34 to 42 degrees C resulted in a 7- to 11-h period in which no growth occurred (compared to wild-type E. coli). These data suggest that biochemical properties of Gpm could be one contributing factor to the heat sensitivity of T. pallidum. PMID:11466272

Benoit, S; Posey, J E; Chenoweth, M R; Gherardini, F C

2001-08-01

131

Treponema pallidum 3-Phosphoglycerate Mutase Is a Heat-Labile Enzyme That May Limit the Maximum Growth Temperature for the Spirochete  

PubMed Central

In the causative agent of syphilis, Treponema pallidum, the gene encoding 3-phosphoglycerate mutase, gpm, is part of a six-gene operon (tro operon) that is regulated by the Mn-dependent repressor TroR. Since substrate-level phosphorylation via the Embden-Meyerhof pathway is the principal way to generate ATP in T. pallidum and Gpm is a key enzyme in this pathway, Mn could exert a regulatory effect on central metabolism in this bacterium. To study this, T. pallidum gpm was cloned, Gpm was purified from Escherichia coli, and antiserum against the recombinant protein was raised. Immunoblots indicated that Gpm was expressed in freshly extracted infective T. pallidum. Enzyme assays indicated that Gpm did not require Mn2+ while 2,3-diphosphoglycerate (DPG) was required for maximum activity. Consistent with these observations, Mn did not copurify with Gpm. The purified Gpm was stable for more than 4 h at 25°C, retained only 50% activity after incubation for 20 min at 34°C or 10 min at 37°C, and was completely inactive after 10 min at 42°C. The temperature effect was attenuated when 1 mM DPG was added to the assay mixture. The recombinant Gpm from pSLB2 complemented E. coli strain PL225 (gpm) and restored growth on minimal glucose medium in a temperature-dependent manner. Increasing the temperature of cultures of E. coli PL225 harboring pSLB2 from 34 to 42°C resulted in a 7- to 11-h period in which no growth occurred (compared to wild-type E. coli). These data suggest that biochemical properties of Gpm could be one contributing factor to the heat sensitivity of T. pallidum. PMID:11466272

Benoit, Stéphane; Posey, James E.; Chenoweth, Matthew R.; Gherardini, Frank C.

2001-01-01

132

Examination of various cell culture techniques for co-incubation of virulent Treponema pallidum (Nichols I strain) under anaerobic conditions.  

PubMed

Treponema pallidum (Nichols virulent) was incubated with and without cells in cell culture medium reduced to -275 mV Ecal, pH 7.3, under deoxygenated conditions. Five to ten percent of the treponemes attached to cells and remained motile for at least 120 h in cell-treponeme systems of co-incubation. Virulent treponemes could be detected after 120 to 144 h in the supernatant fluids of cell-treponeme co-incubation cultures and in cell-free tubes containing medium harvested from aerobically cultivated mammalian cells. Medium supplemented with ox serum ultrafiltrate, pyruvate, and sodium thioglycolate and gas mixtures containing H2 and CO2 enhanced treponemal survival. Increases in treponemal numbers were observed using dark-field microscopy but were not substantiated using the rabbit lesion test. Continuous passage of the treponeme was not achieved in vitro. PMID:789395

Sandok, P L; Knight, S T; Jenkin, H M

1976-10-01

133

Broad specificity AhpC-like peroxiredoxin and its thioredoxin reductant in the sparse antioxidant defense system of Treponema pallidum  

PubMed Central

Little is known about the mechanisms by which Treponema pallidum (Tp), the causative agent of syphilis, copes with oxidative stress as it establishes persistent infection within its obligate human host. The Tp genomic sequence indicates that the bacterium’s antioxidant defenses do not include glutathione and are limited to just a few proteins, with only one, TP0509, offering direct defense against peroxides. Although this Tp peroxiredoxin (Prx) closely resembles AhpC-like Prxs, Tp lacks AhpF, the typical reductant for such enzymes. Functionally, TpAhpC resembles largely eukaryotic, nonAhpC typical 2-Cys Prx proteins in using thioredoxin (Trx, TP0919) as an efficient electron donor and exhibiting broad specificity toward hydroperoxide substrates. Unlike many of the eukaryotic Prxs, however, TpAhpC is relatively resistant to inactivation during turnover with hydroperoxide substrates. As is often observed in typical 2-Cys Prxs, TpAhpC undergoes redox-sensitive oligomer formation. Quantitative immunoblotting revealed that TpTrx and TpAhpC are present at very high levels (over 100 and 300 ?M, respectively) in treponemes infecting rabbit testes; their redox potentials, at -242 ± 1 and -192 ± 2 mV, respectively, are consistent with the role of TpTrx as the cellular reductant of TpAhpC. Transcriptional analysis of select antioxidant genes confirmed the presence of high mRNA levels for ahpC and trx which diminish greatly when spirochetes replicate under in vitro growth conditions. Thus, T. pallidum has evolved an extraordinarily robust, broad-spectrum AhpC as its sole mechanism for peroxide defense to combat this significant threat to treponemal growth and survival during infection. PMID:20304799

Parsonage, Derek; Desrosiers, Daniel C.; Hazlett, Karsten R. O.; Sun, Yongcheng; Nelson, Kimberly J.; Cox, David L.; Radolf, Justin D.; Poole, Leslie B.

2010-01-01

134

Multicentre surveillance of prevalence of the 23S rRNA A2058G and A2059G point mutations and molecular subtypes of Treponema pallidum in Taiwan, 2009-2013.  

PubMed

Resistance mutations A2058G and A2059G, within the 23S rRNA gene of Treponema pallidum, have been reported to cause treatment failures in patients receiving azithromycin for syphilis. Genotyping of T. pallidum strains sequentially isolated from patients with recurrent syphilis is rarely performed. From September 2009 to August 2013, we collected 658 clinical specimens from 375 patients who presented with syphilis for genotyping to examine the number of 60-bp repeats in the acidic repeat protein (arp) gene, T. pallidum repeat (tpr) polymorphism, and tp0548 gene, and to detect A2058G and A2059G point mutations by restriction fragment length polymorphism. Treponemal DNA was identified in 45.2% (n = 298) of the specimens that were collected from 216 (57.6%) patients; 268 (40.7%) specimens tested positive for the 23S rRNA gene, and were examined for macrolide resistance. Two isolates (0.7%) harboured the A2058G mutation, and no A2059G mutation was identified. A total of 14 strains of T. pallidum were identified, with 14f/f (57.5%) and 14b/c (10.0%) being the two predominant strains. Forty patients who presented with recurrent episodes of syphilis had T. pallidum DNA identified from the initial and subsequent episodes, with five cases showing strain discrepancies. One patient had two strains identified from different clinical specimens collected in the same episode. Our findings show that 14f/f is the most common T. pallidum strain in Taiwan, where the prevalence of T. pallidum strains that show A2058G or A2059G mutation remains low. Different genotypes of T. pallidum can be identified in patients with recurrent episodes of syphilis. PMID:24438059

Wu, B-R; Yang, C-J; Tsai, M-S; Lee, K-Y; Lee, N-Y; Huang, W-C; Wu, H; Lee, C-H; Chen, T-C; Ko, W-C; Lin, H-H; Lu, P-L; Chen, Y-H; Liu, W-C; Yang, S-P; Wu, P-Y; Su, Y-C; Hung, C-C; Chang, S-Y

2014-08-01

135

Identification of homologs for thioredoxin, peptidyl prolyl cis-trans isomerase, and glycerophosphodiester phosphodiesterase in outer membrane fractions from Treponema pallidum, the syphilis spirochete.  

PubMed

In this study, we characterized candidate rare outer membrane (OM) proteins with apparent molecular masses of 19, 27, 38, and 38.5 kDa, which had been identified previously in OM fractions from Treponema pallidum (J. D. Radolf et al., Infect. Immun. 63:4244-4252, 1995). Using N-terminal and internal amino acid sequences, a probe for the 19-kDa candidate was PCR amplified and used to screen a T. pallidum genomic library in Lambda Zap II. The corresponding gene (tlp) encoded a homolog for periplasmic thioredoxin-like proteins (Tlp), which reduce c-type cytochromes. A degenerate oligonucleotide derived from the N terminus of the 27-kDa protein was used to PCR amplify a duplex probe from a T. pallidum genomic library in pBluescript II SK+. With this probe, the corresponding gene (ppiB) was identified and found to code for a presumptive periplasmic cyclophilin B-type peptidyl prolyl cis-trans isomerase (PpiB). We postulate that PpiB assists the folding of proteins within the T. pallidum periplasmic space. The N terminus of the 38-kDa candidate was blocked to Edman degradation. However, internal sequence data revealed that it was basic membrane protein (Bmp), a previously characterized, signal peptidase I-processed protein. Triton X-114 phase partitioning revealed that despite its name, Bmp is hydrophilic and therefore likely to be periplasmic. The final candidate was also blocked to Edman degradation; as before, a duplex probe was PCR amplified with degenerate primers derived from internal sequences. The corresponding gene (glpQ) coded for a presumptively lipid-modified homolog of glycerophosphodiester phosphodiesterase (GlpQ). Based upon findings with other treponemal lipoproteins, the hydrophilic GlpQ polypeptide is thought to be anchored by N-terminal lipids to the periplasmic leaflet(s) of the cytoplasmic membrane and/or OM. The discovery of T. pallidum periplasmic proteins with potentially defined functions provides fresh insights into a poorly understood aspect of treponemal physiology. At the same time, however, these findings also raise important issues regarding the use of OM preparations for identifying rare OM proteins of T. pallidum. PMID:9317025

Shevchenko, D V; Akins, D R; Robinson, E J; Li, M; Shevchenko, O V; Radolf, J D

1997-10-01

136

Introduction to Antibodies  

NSDL National Science Digital Library

This site contains a thorough overview of the fundamentals of antibodies. The site starts with an introduction to antigens and antibodies, antibody production and titer including practical information.

2011-02-14

137

Whole Genome Sequence of the Treponema pallidum subsp. endemicum Strain Bosnia A: The Genome Is Related to Yaws Treponemes but Contains Few Loci Similar to Syphilis Treponemes  

PubMed Central

Background T. pallidum subsp. endemicum (TEN) is the causative agent of bejel (also known as endemic syphilis). Clinical symptoms of syphilis and bejel are overlapping and the epidemiological context is important for correct diagnosis of both diseases. In contrast to syphilis, caused by T. pallidum subsp. pallidum (TPA), TEN infections are usually spread by direct contact or contaminated utensils rather than by sexual contact. Bejel is most often seen in western Africa and in the Middle East. The strain Bosnia A was isolated in 1950 in Bosnia, southern Europe. Methodology/Principal Findings The complete genome of the Bosnia A strain was amplified and sequenced using the pooled segment genome sequencing (PSGS) method and a combination of three next-generation sequencing techniques (SOLiD, Roche 454, and Illumina). Using this approach, a total combined average genome coverage of 513× was achieved. The size of the Bosnia A genome was found to be 1,137,653 bp, i.e. 1.6–2.8 kbp shorter than any previously published genomes of uncultivable pathogenic treponemes. Conserved gene synteny was found in the Bosnia A genome compared to other sequenced syphilis and yaws treponemes. The TEN Bosnia A genome was distinct but very similar to the genome of yaws-causing T. pallidum subsp. pertenue (TPE) strains. Interestingly, the TEN Bosnia A genome was found to contain several sequences, which so far, have been uniquely identified only in syphilis treponemes. Conclusions/Significance The genome of TEN Bosnia A contains several sequences thought to be unique to TPA strains; these sequences very likely represent remnants of recombination events during the evolution of TEN treponemes. This finding emphasizes a possible role of repeated horizontal gene transfer between treponemal subspecies in shaping the Bosnia A genome. PMID:25375929

Zobanikova, Marie; Cejkova, Darina; Fulton, Lucinda L.; Chen, Lei; Giacani, Lorenzo; Centurion-Lara, Arturo; Bruisten, Sylvia M.; Sodergren, Erica; Weinstock, George M.; Smajs, David

2014-01-01

138

Simultaneous PCR Detection ofHaemophilus ducreyi,Treponema pallidum, and Herpes Simplex Virus Types 1 and 2 from Genital Ulcers  

Microsoft Academic Search

A multiplex PCR (M-PCR) assay with colorimetric detection was devised for the simultaneous amplification of DNA targets fromHaemophilus ducreyi,Treponema pallidum, and herpes simplex virus (HSV) types 1 and 2. By using target-specific oligonucleotides in a microwell format, 298 genital ulcer swab specimens collected in New Orleans during three intervals from 1992 through 1994 were evaluated. The results of the M-PCR

KARINA A. ORLE; CAROL A. GATES; DAVID H. MARTIN; BARBARA A. BODY; ANDJUDITH B. WEISS

1996-01-01

139

Syphilis-causing strains belong to separate SS14-like or Nichols-like groups as defined by multilocus analysis of 19 Treponema pallidum strains.  

PubMed

Treponema pallidum strains are closely related at the genome level but cause distinct diseases. Subspecies pallidum (TPA) is the causative agent of syphilis, subspecies pertenue (TPE) causes yaws while subspecies endemicum (TEN) causes bejel (endemic syphilis). Compared to the majority of treponemal genomic regions, several chromosomal loci were found to be more diverse. To assess genetic variability in diverse genomic positions, we have selected (based on published genomic data) and sequenced five variable loci, TP0304, TP0346, TP0488, TP0515 and TP0558, in 19 reference Treponema pallidum strains including all T. pallidum subspecies (TPA, TPE and TEN). Results of this multilocus analysis divided syphilitic isolates into two groups: SS14-like and Nichols-like. The SS14-like group is comprised of SS14, Grady, Mexico A and Philadelphia 1 strains. The Nichols-like group consisted of strains Nichols, Bal 73-1, DAL-1, MN-3, Philadelphia 2, Haiti B and Madras. The TP0558 locus was selected for further studies because it clearly distinguished between the SS14- and Nichols-like groups and because the phylogenetic tree derived from the TP0558 locus showed the same clustering pattern as the tree constructed from whole genome sequences. In addition, TP0558 was shown as the only tested locus that evolved under negative selection within TPA strains. Sequencing of a short fragment (573bp) of the TP0558 locus in a set of 25 clinical isolates from 22 patients collected in the Czech Republic during 2012-2013 revealed that clinical isolates follow the SS14- and Nichols-like distribution. PMID:24841252

Nechvátal, Lukáš; P?trošová, Helena; Grillová, Linda; Pospíšilová, Petra; Mikalová, Lenka; Strnadel, Radim; Kuklová, Ivana; Kojanová, Martina; Kreidlová, Miluše; Va?ousová, Daniela; Procházka, P?emysl; Zákoucká, Hana; Krch?áková, Alena; Smajs, David

2014-07-01

140

Immunization with the N-Terminal Portion of Treponema pallidum Repeat Protein K Attenuates Syphilitic Lesion Development in the Rabbit Model  

Microsoft Academic Search

When used as an immunogen, Treponema pallidum repeat protein K (TprK) has been shown to attenuate syphilitic lesions upon homologous intradermal challenge in the rabbit model. To further explore this protein as a potential vaccine component, we sought to identify the immunogenic regions of TprK. The abilities of three recombinant peptides encompassing TprK to elicit T- and B-cell responses and

Cecilia A. Morgan; Sheila A. Lukehart; Wesley C. Van Voorhis

2002-01-01

141

Antimitochondrial antibodies  

Microsoft Academic Search

Laboratory-prepared and commercially obtained fluorescein-labeled rabbit antihuman IgG were compared in performing the antimitochondrial antibody (AMA) assay. Identical results were obtained using either of the fluorescent antisera at protein concentrations of 1.5 mg\\/ml and 1:10 dilutions of patients' sera. Positive AMA tests with either antisera were observed in each of 7 patients with primary biliary cirrhosis (PBC), 2 of 83

Stephen L. Winter; Sumner C. Kraft; James L. Boyer

1979-01-01

142

New Tests for Syphilis: Rational Design of a PCR Method for Detection of Treponema pallidum in Clinical Specimens Using Unique Regions of the DNA Polymerase I Gene  

PubMed Central

A sensitive and specific PCR method to detect Treponema pallidum in clinical specimens was developed. PCR primers were designed based on two unique features of the DNA polymerase I gene (polA). The first distinctive characteristic is that the region codes for a high cysteine content and has low homology with similar regions of DNA polymerase I gene from known microorganisms. The second unique feature is the presence of four insertions in the gene. PCR tests using primers designed on the basis these regions reacted with various pathogenic T. pallidum subspecies but did not react with nonpathogenic treponemal species or other spirochetes. An additional 59 species of bacteria and viruses, including those that cause genital ulcers, tested negative. This PCR method is extremely robust and sensitive. The detection limit is about 10 to 25 organisms when analyzed on gel. However, the analytic sensitivity can be increased by at least 1 log, to a detection limit of a single organism, when the ABI 310 Prism Genetic Analyzer is used to detect fluorescence-labeled amplicons. We further used this test in a clinical setting and compared the results with results from a previously reported multiplex-PCR test (for T. pallidum, Haemophilus ducreyi, and herpes simplex virus). We tested 112 genital ulcer specimens by the polA PCR, obtaining a sensitivity of 95.8% and a specificity of 95.7%. These results suggest that the polA PCR is applicable as a routine clinical diagnostic test for syphilis. PMID:11326018

Liu, Hsi; Rodes, Berta; Chen, C.-Y.; Steiner, Bret

2001-01-01

143

Malignant syphilis with ocular involvement in an HIV-infected patient.  

PubMed

Malignant syphilis is now considered a rare disease, more commonly affecting individuals with poor health, malnutrition or HIV infection. We present a 34-year-old man with HIV infection who developed multiple atypical cutaneous ulcerations, leonine facies, a scleral nodule and keratitis with visual loss. The diagnosis of malignant syphilis was delayed due to the insidious presentation, but was confirmed via immunohistochemical (IHC) staining with anti-Treponema antibodies of a skin biopsy. Significant clinical improvement was observed following a 15-day course of penicillin and tigecycline therapy. In advanced HIV disease, cutaneous manifestations are often difficult to identify and present a challenge for the clinician. Clinical manifestations of secondary syphilis vary greatly, earning the epigram of 'the great imitator'. It is important to recognize atypical presentations of syphilis, especially among HIV-infected individuals. Unlike historical cases of malignant syphilis, Treponema pallidum was found in the tissue section using IHC staining methods. We emphasize the importance of lues maligna in the differential diagnosis of HIV-infected patients with diffuse ulceronodular lesions as well as the usefulness of histological investigations and IHC studies. PMID:21571984

De Socio, G V L; Simonetti, S; Tomasini, C; Ansidei, V; Pasticci, M B; Baldelli, F

2011-05-01

144

The Social Amoeba Polysphondylium pallidum Loses Encystation and Sporulation, but Can Still Erect Fruiting Bodies in the Absence of Cellulose  

PubMed Central

Amoebas and other freely moving protists differentiate into walled cysts when exposed to stress. As cysts, amoeba pathogens are resistant to biocides, preventing treatment and eradication. Lack of gene modification procedures has left the mechanisms of encystation largely unexplored. Genetically tractable Dictyostelium discoideum amoebas require cellulose synthase for formation of multicellular fructifications with cellulose-rich stalk and spore cells. Amoebas of its distant relative Polysphondylium pallidum (Ppal), can additionally encyst individually in response to stress. Ppal has two cellulose synthase genes, DcsA and DcsB, which we deleted individually and in combination. Dcsa- mutants formed fruiting bodies with normal stalks, but their spore and cyst walls lacked cellulose, which obliterated stress-resistance of spores and rendered cysts entirely non-viable. A dcsa-/dcsb- mutant made no walled spores, stalk cells or cysts, although simple fruiting structures were formed with a droplet of amoeboid cells resting on an sheathed column of decaying cells. DcsB is expressed in prestalk and stalk cells, while DcsA is additionally expressed in spores and cysts. We conclude that cellulose is essential for encystation and that cellulose synthase may be a suitable target for drugs to prevent encystation and render amoeba pathogens susceptible to conventional antibiotics. PMID:25113829

Du, Qingyou; Schaap, Pauline

2014-01-01

145

Efficient preparation and metal specificity of the regulatory protein TroR from the human pathogen Treponema pallidum.  

PubMed

TroR is a putative metal-dependent regulatory protein that has been linked to the virulence of the human pathogen Treponema pallidum. It shares high homology with the well-known iron-dependent regulatory protein DtxR from Corynebacterium diphtheriae, as well as the manganese-dependent MntR from Bacillus subtilis. However, it has been uncertain whether manganese or zinc is the natural cofactor of TroR to date. Herein, we established an efficient method named "double-fusion tagging" to obtain soluble TroR for the first time. A series of studies, including ICP, CD, fluorescence, ITC, and electrophoresis mobility shift assay (EMSA), were performed to resolve the discrepancies in its metal-binding specificity. In addition, bioinformatic analysis as well as mutation studies were carried out to find the genetic relationships of TroR with its homology proteins. In conclusion, our findings indicate that TroR is a manganese-dependent rather than a zinc-dependent regulatory protein. PMID:23945957

Liu, Yi; Li, Wei; Wei, Yaozhu; Jiang, Yindi; Tan, Xiangshi

2013-10-01

146

The rostral subcommissural ventral pallidum is a mix of ventral pallidal neurons and neurons from adjacent areas: an electrophysiological study  

PubMed Central

The ventral pallidum (VP) is a part of the ventral striatopallidal system and is involved in reward-related behaviors. The VP is composed of a ventromedial (VPvm) and a dorsolateral (VPdl) subregion, and some rostral-caudal differences are reported. Study of the VP often focuses on the subcommissural VP, typically considered homogenous in spite of known subdivisions. In this work, we used slice electrophysiology combined with immunohistochemistry for marker neuropeptides to test whether the subcommissural VP is functionally homogenous. Using sagittal slices, we show that more lateral levels (2.40 mm) of the subcommissural VP are homogenous but that a more medial slice (1.90 mm) contains two types of neurons. One type, located more caudally, resembles neurons in the lateral subcommissural VP, with long aspiny dendrites, primarily GABAergic input, and characteristic electrophysiological properties, such as depolarized membrane potential and spontaneous action potential discharge. The second type of neuron, located mostly in the rostral subcommissural VP, shows properties that are akin to medium spiny neurons of adjacent regions, including spiny dendrites, major glutamatergic input, hyperpolarized membrane potential, and no spontaneous action potentials. The two types of neurons were present in both the VPvm and VPdl, implying that the mix is not a characteristic of histologically defined subregions. We conclude that at medial levels the rostral subcommissural VP contains a mix of typical ventral pallidal neurons and spiny neurons similar to those in adjacent regions. This observation needs to be considered when interpreting past experiments and designing future experiments in the subcommissural VP. PMID:23143342

Kalivas, Peter W.

2012-01-01

147

Dopamine receptor regulation of ethanol intake and extracellular dopamine levels in the ventral pallidum of alcohol preferring (P) rats.  

PubMed

Sufficient evidence exists for the inclusion of the ventral pallidum (VP) into the category of a dopaminoceptive brain region. The effects of inhibiting dopamine D(1)- or D(2)-like receptors in the VP on (a) ethanol intake and (b) extracellular levels of dopamine, were investigated in the alcohol-preferring (P) rat. The D(1)-like antagonist, SCH-23390, and the D(2)-like antagonist, sulpiride (0.25-2 microg/0.5 microl) were bilaterally injected into the VP and ethanol (15%, v/v) intake was assessed in a 1 h limited access paradigm. The results indicate that microinjections of sulpiride significantly increased ethanol consumption (65% increase at the 2.0 microg dose). Whereas the D(1) antagonists SCH-23390 tended to decrease ethanol intake, the effect was not statistically significant. In a separate group of rats, reverse microdialysis of sulpiride and SCH-23390 (10-200 microM) were conducted in the VP of P rats. Local perfusion of only the 200 microM sulpiride dose significantly increased the extracellular levels of dopamine (maximal increase: 250% of baseline). On the other hand, local perfusion of SCH-23390 (10-200 microM) dose dependently increased the extracellular levels of dopamine 180-640% of baseline. Overall, the results of this study suggest that (a) tonic activation of D(2) postsynaptic receptors in VP imposes a limit on ethanol intake in the P rat; (b) there are few D(2) autoreceptors functioning in the VP; (c) there is tonic D(1)-like receptor mediated inhibitory feedback regulation of VP-dopamine release. PMID:15734229

Melendez, Roberto I; Rodd, Zachary A; McBride, William J; Murphy, James M

2005-03-01

148

Manipulation of GABA in the ventral pallidum, but not the nucleus accumbens, induces intense, preferential, fat consumption in rats.  

PubMed

Injections of the GABAA antagonist bicuculline into the medial ventral pallidum (VPm) induce marked increases in food intake, but nothing is known about the way in which these injections alter the distribution of intake in a macronutrient selection situation. We investigated this topic by adapting rats to a diet containing independent sources of protein, carbohydrate and fat, and then examining the effects of intra-VPm bicuculline on diet selection. Under these conditions, bicuculline produced a massive, preferential increase in fat intake with subjects consuming a mean of 97% of their calories from fat. Furthermore, all treated subjects ate fat before any other macronutrient, suggesting that the animals' behavior was directed selectively toward this dietary component even before consumption had begun. Similar effects were not observed following food deprivation, which exerted its largest effect on carbohydrate intake. To compare the intra-VPm bicuculline response to that seen after activation of GABA receptors in the nucleus accumbens shell (AcbSh), a major source of projections to the VPm, we conducted similar experiments with intra-AcbSh injections of muscimol and baclofen. These injections also enhanced food intake, but did not reproduce the selective preference for fat seen after intra-VPm bicuculline. These experiments provide the first demonstration of preferential enhancement of fat intake following manipulations of a nonpeptide neurotransmitter. Since mean intakes of fat under baseline conditions and after deprivation tended to be lower than those of carbohydrates, it seems unlikely that the effects of intra-VPm bicuculline are related to the intrinsic "rewarding" properties of fat, but might rather reflect the induction of a state of "fat craving." PMID:24867334

Covelo, Ignacio R; Patel, Zaid I; Luviano, Jennifer A; Stratford, Thomas R; Wirtshafter, David

2014-08-15

149

Ability of Macrophages to Process and Present Treponema pallidum Bosnia A Strain Antigens in Experimental Syphilis of Syrian Hamsters  

PubMed Central

The ability of macrophages to process and present treponemal antigens to T-lymphocytes was studied in early stages of experimental syphilis produced by Treponema pallidum Bosnia A strain (the causative agent of endemic syphilis) infection of inbred Syrian hamsters (LSH/Ss Lak strain). A difference was noticed in the response of macrophages obtained from the peritoneal cavity, lymph nodes, and spleens of the infected animals. In all of these locations, a general increase in the population of Iak-positive macrophage was seen during the entire period of infection, i.e., 3 to 18 weeks after inoculation. Peritoneal cavity-derived macrophages showed no difference in antigen presentation to sensitized and nonsensitized T-lymphocytes for the first 7 weeks of infection. However, at 18 weeks after infection, peritoneal macrophages lost their ability to process treponema antigens. Spleen- and lymph node-derived macrophages did not exhibit a parallel loss in their ability to process treponema antigens. A fluctuation without a consistent pattern was noticed in the antigen processing and presentation by macrophages from the spleen and lymph nodes. In general, the sensitized T-lymphocytes responded to treponema antigen presented by macrophages more vigorously than the nonsensitized T-lymphocytes. An increased ability of spleen-derived macrophages to process and present antigens was noticed throughout the entire period of infection. The macrophages from the lymph nodes showed such an increase only temporarily at 3 weeks after infection. These data suggest that the processing and presentation of treponema antigens by macrophages in acute syphilitic infection fluctuates considerably and depends on the source of macrophages and the duration of the infection. The differences in the response of peritoneal cavity-, spleen-, and lymph node-derived macrophages probably reflect the complex interactions between the macrophage and other cells involved in the immune response to treponema infection. PMID:7042569

Bagasra, Omar; Damjanov, Ivan

1982-01-01

150

Vitros 5600 Syphilis TPA Assay: Evaluation of an Automated Chemiluminescence Assay for Detection of Treponema pallidum Antibodies in a High Prevalence Setting.  

PubMed

The performance of the Syphilis TPA assay (Ortho-Clinical Diagnostics) on Vitros 5600 Integrated System was evaluated and demonstrated excellent results. Our data support the use of this assay for test confirmation in the traditional algorithm and for screening for syphilis in a routine automated laboratory setting when using the reverse algorithm. PMID:25299416

Van den Bossche, Dorien; Florence, Eric; Kenyon, Christopher; Van Esbroeck, Marjan

2014-11-01

151

The TP0796 Lipoprotein of Treponema pallidum Is a Bimetal-dependent FAD Pyrophosphatase with a Potential Role in Flavin Homeostasis*  

PubMed Central

Treponema pallidum, an obligate parasite of humans and the causative agent of syphilis, has evolved the capacity to exploit host-derived metabolites for its survival. Flavin-containing compounds are essential cofactors that are required for metabolic processes in all living organisms, and riboflavin is a direct precursor of the cofactors FMN and FAD. Unlike many pathogenic bacteria, Treponema pallidum cannot synthesize riboflavin; we recently described a flavin-uptake mechanism composed of an ABC-type transporter. However, there is a paucity of information about flavin utilization in bacterial periplasms. Using a discovery-driven approach, we have identified the TP0796 lipoprotein as a previously uncharacterized Mg2+-dependent FAD pyrophosphatase within the ApbE superfamily. TP0796 probably plays a central role in flavin turnover by hydrolyzing exogenously acquired FAD, yielding AMP and FMN. Biochemical and structural investigations revealed that the enzyme has a unique bimetal Mg2+ catalytic center. Furthermore, the pyrophosphatase activity is product-inhibited by AMP, indicating a possible role for this molecule in modulating FMN and FAD levels in the treponemal periplasm. The ApbE superfamily was previously thought to be involved in thiamine biosynthesis, but our characterization of TP0796 prompts a renaming of this superfamily as a periplasmic flavin-trafficking protein (Ftp). TP0796 is the first structurally and biochemically characterized FAD pyrophosphate enzyme in bacteria. This new paradigm for a bacterial flavin utilization pathway may prove to be useful for future inhibitor design. PMID:23447540

Deka, Ranjit K.; Brautigam, Chad A.; Liu, Wei Z.; Tomchick, Diana R.; Norgard, Michael V.

2013-01-01

152

The Treponema pallidum tro operon encodes a multiple metal transporter, a zinc-dependent transcriptional repressor, and a semi-autonomously expressed phosphoglycerate mutase.  

PubMed

The Treponema pallidum tro operon encodes an ABC transporter (TroABCD), a transcriptional repressor (TroR), and the essential glycolytic enzyme phosphoglycerate mutase (Gpm). The apparently discordant observations that the solute binding protein (TroA) binds Zn2+, whereas DNA binding by TroR in vitro is Mn2+-dependent, have generated uncertainty regarding the identities of the ligand(s) and co-repressor(s) of the permease. Moreover, this operonic structure suggests that Gpm expression, and hence glycolysis, the sole source of ATP for the bacterium, would be suspended during TroR-mediated repression. To resolve these discrepancies, we devised an experimental strategy permitting a more direct assessment of Tro operon function and regulation. We report that (i) apo-TroA has identical affinities for Zn2+ and Mn2+; (ii) the Tro transporter expressed in Escherichia coli imports Zn2+, Mn2+, and possibly iron; (iii) TroR represses transporter expression in E. coli at significantly lower concentrations of Zn2+ than of Mn2+; and (iv) TroR-mediated repression causes a disproportionately greater down-regulation of the transporter genes than of gpm. The much higher concentrations of Zn2+ than of Mn2+ in human body fluids suggests that Zn2+ is both the primary substrate and co-repressor of the permease in vivo. Our data also indicate that Gpm expression and, therefore, glycolysis would not be abrogated when T. pallidum encounters high Zn2+ levels. PMID:12668673

Hazlett, Karsten R O; Rusnak, Frank; Kehres, David G; Bearden, Scott W; La Vake, Carson J; La Vake, Morgan E; Maguire, Michael E; Perry, Robert D; Radolf, Justin D

2003-06-01

153

The multifunctional role of the pallilysin-associated Treponema pallidum protein, Tp0750, in promoting fibrinolysis and extracellular matrix component degradation.  

PubMed

The mechanisms that facilitate dissemination of the highly invasive spirochaete, Treponema pallidum, are incompletely understood. Previous studies showed the treponemal metalloprotease pallilysin (Tp0751) possesses fibrin clot degradation capability, suggesting a role in treponemal dissemination. In the current study we report characterization of the functionally linked protein Tp0750. Structural modelling predicts Tp0750 contains a von Willebrand factor type A (vWFA) domain, a protein-protein interaction domain commonly observed in extracellular matrix (ECM)-binding proteins. We report Tp0750 is a serine protease that degrades the major clot components fibrinogen and fibronectin. We also demonstrate Tp0750 cleaves a matrix metalloprotease (MMP) peptide substrate that is targeted by several MMPs, enzymes central to ECM remodelling. Through proteomic analyses we show Tp0750 binds the endothelial fibrinolytic receptor, annexin A2, in a specific and dose-dependent manner. These results suggest Tp0750 constitutes a multifunctional protein that is able to (1) degrade infection-limiting clots by both inhibiting clot formation through degradation of host coagulation cascade proteins and promoting clot dissolution by complexing with host proteins involved in the fibrinolytic cascade and (2) facilitate ECM degradation via MMP-like proteolysis of host components. We propose that through these activities Tp0750 functions in concert with pallilysin to enable T.?pallidum dissemination. PMID:24303899

Houston, Simon; Russell, Shannon; Hof, Rebecca; Roberts, Alanna K; Cullen, Paul; Irvine, Kyle; Smith, Derek S; Borchers, Christoph H; Tonkin, Michelle L; Boulanger, Martin J; Cameron, Caroline E

2014-02-01

154

Molecular Characterization of Syphilis in Patients in Canada: Azithromycin Resistance and Detection of Treponema pallidum DNA in Whole-Blood Samples versus Ulcerative Swabs?  

PubMed Central

Although detection of Treponema pallidum DNA in whole-blood specimens of syphilis patients has been reported, it is uncertain at what stage of the disease such specimens are most suitable for the molecular diagnosis of syphilis. Also, few studies have directly compared the different gene targets for routine laboratory diagnostic usage in PCR assays. We examined 87 specimens from 68 patients attending two urban sexually transmitted disease clinics in Alberta, Canada. PCR was used to amplify the T. pallidum tpp47, bmp, and polA genes as well as a specific region of the 23S rRNA gene linked to macrolide antibiotic susceptibility. In primary syphilis cases, PCR was positive exclusively (75% sensitivity rate) in ulcerative swabs but not in blood specimens, while in secondary syphilis cases, 50% of the blood specimens were positive by PCR. Four out of 14 (28.6%) of our PCR-positive syphilis cases were found to be caused by an azithromycin-resistant strain(s). Our results confirmed that swabs from primary ulcers are the specimens of choice for laboratory diagnostic purposes. However, further research is required to determine what specimen(s) would be most appropriate for molecular investigation of syphilis in secondary and latent syphilis. PMID:19339468

Martin, Irene E.; Tsang, Raymond S. W.; Sutherland, Karen; Tilley, Peter; Read, Ron; Anderson, Barbara; Roy, Colleen; Singh, Ameeta E.

2009-01-01

155

[Prevalence, risk factors and genetic characterization of human T-cell lymphotropic virus types 1 and 2 in patients infected with human immunodeficiency virus type 1 in the cities of Ribeirăo Preto and Săo Paulo].  

PubMed

The aim of this study was to define the prevalence of human T cell lymphotropic virus types 1 and 2 in patients who were positive for human immunodeficiency virus type 1 in the State of Săo Paulo, Brazil. We evaluated 319 individuals infected with HIV type 1 who were attended at specialized clinics in two cities (Ribeirăo Preto and Săo Paulo). The patients were interviewed and tested for antibodies against HTLV types 1 and 2 (Orthoâ HTLV-1/HTLV-2 Ab-Capture enzyme immunoassay). Direct DNA sequencing of polymerase chain reaction products from the tax region of HTLV type 2 and the long terminal repeat region of HTLV types 1 and 2 were performed to differentiate and determine the subtypes. The overall prevalence of anti-HTLV type 1 and 2 antibodies was 7.5% (24/319; 95% CI: 5.2-11.5). HTLV type 1 and 2 infection was associated with a history of injected drug use and with antibodies for hepatitis C virus (p < 0.001), but not with age (p = 0.2), sex (p = 0.9), sexual behavior or serological markers for sexually transmitted diseases (anti-Treponema pallidum, anti-human herpesvirus type 8 or anti-hepatitis B virus antibodies) (p > 0.05). HTLV DNA was detected in 13 out of 24 samples, of which 12 were characterized as HTLV subtype 2c and one as HTLV subtype 1a. Among the 12 HTLV type 2 samples, seven were from injected drug users, thus indicating that this route is an important risk factor for HTLV type 2 transmission among our population infected with HIV type 1. PMID:19684973

Kleine Neto, Walter; Sanabani, Sabri Saeed; Jamal, Leda Fátima; Sabino, Ester Cerdeira

2009-01-01

156

Renaturation of Recombinant Treponema pallidum Rare Outer Membrane Protein 1 into a Trimeric, Hydrophobic, and Porin-Active Conformation  

PubMed Central

We have previously observed that while native Treponema pallidum rare outer membrane protein 1 (Tromp1) is hydrophobic and has porin activity, recombinant forms of Tromp1 do not possess these properties. In this study we show that these properties are determined by conformation and can be replicated by proper renaturation of recombinant Tromp1. Native Tromp1, but not the 47-kDa lipoprotein, extracted from whole organisms by using Triton X-114, was found to lose hydrophobicity after treatment in 8 M urea, indicating that Tromp1’s hydrophobicity is conformation dependent. Native Tromp1 was purified from 0.1% Triton X-100 extracts of whole organisms by fast-performance liquid chromatography (FPLC) and shown to have porin activity in planar lipid bilayers. Cross-linking studies of purified native Tromp1 with an 11 ? cross-linking agent showed oligomeric forms consistent with dimers and trimers. For renaturation studies of recombinant Tromp1 (rTromp1), a 31,109-Da signal-less construct was expressed in Escherichia coli and purified by FPLC. FPLC-purified rTromp1 was denatured in 8 M urea and then renatured in the presence of 0.5% Zwittergent 3,14 during dialysis to remove the urea. Renatured rTromp1 was passed through a Sephacryl S-300 gel exclusion column previously calibrated with known molecular weight standards. While all nonrenatured rTromp1 eluted from the column at approximately the position of the carbonic anhydrase protein standard (29 kDa), all renatured rTromp1 eluted at the position of the phosphorylase b protein standard (97 kDa), suggesting a trimeric conformation. Trimerization was confirmed by using an 11 ? cross-linking agent which showed both dimers and trimers similar to that of native Tromp1. Triton X-114 phase separations showed that all of renatured rTromp1, but none of nonrenatured rTromp1, phase separated exclusively into the hydrophobic detergent phase, similar to native Tromp1. Circular dichroism of nonrenatured and renatured rTromp1 showed a marked loss in alpha-helical secondary structure of renatured rTromp1 compared to the nonrenatured form. Finally, renatured rTromp1, but not the nonrenatured form, showed porin activity in planar liquid bilayers. These results demonstrate that proper folding of rTromp1 results in a trimeric, hydrophobic, and porin-active conformation similar to that of the native protein. PMID:10572117

Zhang, Hongwei H.; Blanco, David R.; Exner, Maurice M.; Shang, Ellen S.; Champion, Cheryl I.; Phillips, Martin L.; Miller, James N.; Lovett, Michael A.

1999-01-01

157

Detection of the A2058G and A2059G 23S rRNA Gene Point Mutations Associated with Azithromycin Resistance in Treponema pallidum by Use of a TaqMan Real-Time Multiplex PCR Assay  

PubMed Central

Macrolide treatment failure in syphilis patients is associated with a single point mutation (either A2058G or A2059G) in both copies of the 23S rRNA gene in Treponema pallidum strains. The conventional method for the detection of both point mutations uses nested PCR combined with restriction enzyme digestions, which is laborious and time-consuming. We initially developed a TaqMan-based real-time duplex PCR assay for detection of the A2058G mutation, and upon discovery of the A2059G mutation, we modified the assay into a triplex format to simultaneously detect both mutations. The point mutations detected by the real-time triplex PCR were confirmed by pyrosequencing. A total of 129 specimens PCR positive for T. pallidum that were obtained from an azithromycin resistance surveillance study conducted in the United States were analyzed. Sixty-six (51.2%) of the 129 samples with the A2058G mutation were identified by both real-time PCR assays. Of the remaining 63 samples that were identified as having a macrolide-susceptible genotype by the duplex PCR assay, 17 (27%) were found to contain the A2059G mutation by the triplex PCR. The proportions of macrolide-susceptible versus -resistant genotypes harboring either the A2058G or the A2059G mutation among the T. pallidum strains were 35.6, 51.2, and 13.2%, respectively. None of the T. pallidum strains examined had both point mutations. The TaqMan-based real-time triplex PCR assay offers an alternative to conventional nested PCR and restriction fragment length polymorphism analyses for the rapid detection of both point mutations associated with macrolide resistance in T. pallidum. PMID:23284026

Chi, Kai-Hua; Pillay, Allan; Nachamkin, Eli; Su, John R.; Ballard, Ronald C.

2013-01-01

158

Antibody Blood Tests  

MedlinePLUS

... CeliacDisease.net People with celiac disease who eat gluten have higher than normal levels of certain antibodies ... rye and barley that are generically known as “gluten.” Find Out For Sure Antibody tests are only ...

159

Antiphospholipid Antibody Syndrome  

MedlinePLUS

... fo-LIP-id) antibody syndrome (APS) is an autoimmune disorder. Autoimmune disorders occur if the body's immune system makes antibodies ... long term. If you have APS and another autoimmune disorder, it's important to control that condition as well. ...

160

Serum antibodies to whole-cell and recombinant antigens of Borrelia burgdorferi in cottontail rabbits.  

PubMed

Archived serum samples, from 95 eastern cottontail rabbits (Sylvilagus floridanus) captured in New York, New York, USA and Millbrook, New York, USA, during 1985-86, were analyzed in solid-phase enzyme-linked immunosorbent assays (ELISA) for total and class-specific immunoglobulin (Ig) M antibodies to whole-cell or recombinant antigens of Borrelia burgdorferi sensu stricto. Using a polyvalent conjugate, rabbit sera contained antibodies to whole-cell and recombinant antigens (protein [p]35, p37, or VlsE) during different seasons, but there was no reactivity to outer surface protein (Osp)A or OspB. Seventy-six of the 102 sera (75%) analyzed were reactive with one or more of the antigens; 61 of the positive samples (80%) reacted to whole-cell antigens, followed by results for the p35 (58%, 44/76), VlsE (43%, 33/76), and p37 (29%, 22/ 76) antigens. Fifty-eight sera (76%) contained antibodies to the VlsE or p35 antigens with or without reactivity to whole-cell antigens. High antibody titers (?1:2,560) recorded for 52 sera indicate robust antibody production. In analyses for IgM antibodies in an ELISA containing whole-cell antigens, there were 30 positive sera; titers ranged from 1:160 to 1:640. There was minimal cross-reactivity when rabbit antisera to Treponema pallidum or four serovars of Leptospira interrogans were screened against B. burgdorferi antigens. Based on more-specific results, VlsE and p35 antigens appear to be useful markers for detecting possible B. burgdorferi infections. PMID:22247369

Magnarelli, Louis A; Norris, Steven J; Fikrig, Erol

2012-01-01

161

SERUM ANTIBODIES TO WHOLE-CELL AND RECOMBINANT ANTIGENS OF BORRELIA BURGDORFERI IN COTTONTAIL RABBITS  

PubMed Central

Archived serum samples, from 95 eastern cottontail rabbits (Sylvilagus floridanus) captured in New York, New York, USA and Millbrook, New York, USA, during 1985–86, were analyzed in solid-phase enzyme-linked immunosorbent assays (ELISA) for total and class-specific immunoglobulin (Ig) M antibodies to whole-cell or recombinant antigens of Borrelia burgdorferi sensu stricto. Using a polyvalent conjugate, rabbit sera contained antibodies to whole-cell and recombinant antigens (protein [p]35, p37, or VlsE) during different seasons, but there was no reactivity to outer surface protein (Osp)A or OspB. Seventy-six of the 102 sera (75%) analyzed were reactive with one or more of the antigens; 61 of the positive samples (80%) reacted to whole-cell antigens, followed by results for the p35 (58%, 44/76), VlsE (43%, 33/76), and p37 (29%, 22/76) antigens. Fifty-eight sera (76%) contained antibodies to the VlsE or p35 antigens with or without reactivity to whole-cell antigens. High antibody titers (?1:2,560) recorded for 52 sera indicate robust antibody production. In analyses for IgM antibodies in an ELISA containing whole-cell antigens, there were 30 positive sera; titers ranged from 1:160 to 1:640. There was minimal cross-reactivity when rabbit antisera to Treponema pallidum or four serovars of Leptospira interrogans were screened against B. burgdorferi antigens. Based on more-specific results, VlsE and p35 antigens appear to be useful markers for detecting possible B. burgdorferi infections. PMID:22247369

Magnarelli, Louis A.; Norris, Steven J.; Fikrig, Erol

2011-01-01

162

Presence of Borrelia burgdorferi sensu lato antibodies in the serum of patients with abdominal aortic aneurysms.  

PubMed

Infectious agents are likely to play a role in the pathogenesis of chronic inflammatory diseases, including abdominal aortic aneurysms (AAAs). The goal of this study was to determine if Borrelia burgdorferi sensu lato (sl), a microorganism responsible for Lyme disease, is involved in the etiology of AAAs. The presence of serum antibodies against B. burgdorferi sl was measured with enzyme-linked immunosorbent assay (ELISA) and confirmed by Western blotting in 96 AAA and 108 peripheral artery disease (PAD) patients. Polymerase chain reaction (PCR) was used for the detection of Borrelia-specific DNA in the aneurysm wall. Among AAA patients 34% and among PAD patients 16% were seropositive for B. burgdorferi sl antibodies (Fisher's exact test, p = 0.003; odds ratio [OR] 2.79; 95% confidence interval [CI] 1.37-5.85). In the German general population, 3-17% are seropositive for Borrelia antibodies. No Borrelia DNA was detected in the aneurysm wall. Our findings suggest a relationship between AAAs and B. burgdorferi sl. We hypothesize that the underlying mechanism for B. burgdorferi sl in AAA formation is similar to that by the spirochete Treponema pallidum; alternatively, AAAs could develop due to induced autoimmunity via molecular mimicry due to similarities between some of the B. burgdorferi sl proteins and aortic proteins. PMID:21842293

Hinterseher, I; Gäbel, G; Corvinus, F; Lück, C; Saeger, H D; Bergert, H; Tromp, G; Kuivaniemi, H

2012-05-01

163

Antibodies, viruses and vaccines  

Microsoft Academic Search

Neutralizing antibodies are crucial for vaccine-mediated protection against viral diseases. They probably act, in most cases, by blunting the infection, which is then resolved by cellular immunity. The protective effects of neutralizing antibodies can be achieved not only by neutralization of free virus particles, but also by several activities directed against infected cells. In certain instances, non-neutralizing antibodies contribute to

Dennis R. Burton

2002-01-01

164

Modeling Antibody Diversity.  

ERIC Educational Resources Information Center

Understanding antibody structure and function is difficult for many students. The rearrangement of constant and variable regions during antibody differentiation can be effectively simulated using a paper model. Describes a hands-on laboratory exercise which allows students to model antibody diversity using readily available resources. (PVD)

Baker, William P.; Moore, Cathy Ronstadt

1998-01-01

165

Proposal of a new halobacterial genus Natrinema gen. nov., with two species Natrinema pellirubrum nom. nov. and Natrinema pallidum nom. nov.  

PubMed

A phylogenetic analysis of 69 halobacterial 16S rRNA gene sequences has been carried out, integrating data from new isolates, previously described halobacteria and cloned sequences from uncultivated halobacteria. Halobacterium halobium NCIMB 777, Halobacterium trapanicum NCIMB 784 and Halobacterium salinarium NCIMB 786, together with several other strains (strains T5.7, L11 and Halobacterium trapanicum NCIMB 767) constitute a distinct lineage with at least 98.2% sequence similarity. These strains have been incorrectly assigned to the genus Halobacterium. Therefore, based on a variety of taxonomic criteria, it is proposed that Halobacterium salinarium NCIMB 786 is renamed as Natrinema pellirubrum nom. nov., the type species of the new genus Natrinema gen. nov., and that Halobacterium halobium NCIMB 777 and Halobacterium trapanicum NCIMB 784 are renamed as a single species, Natrinema pallidum nom. nov. It was notable that halobacteria closely related to the proposed new genus have been isolated from relatively low-salt environments. PMID:9828420

McGenity, T J; Gemmell, R T; Grant, W D

1998-10-01

166

Spirochetemia due to Treponema pallidum using polymerase-chain-reaction assays in patients with early syphilis: prevalence, associated factors and treatment response.  

PubMed

Between 2009 and 2013, polymerase-chain-reaction assay was used to detect Treponema pallidum in the blood samples collected from 296 patients with early syphilis (241 being HIV infected) and 102 patients (34.5%) had spirochetemia. The presence of spirochetemia was associated with lower CD4 counts (per 10-cell/mm(3) decrease, adjusted odds ratio (AOR), 1.020; 95% CI, 1.006-1.036) and secondary syphilis (AOR, 4.967; 95% CI, 2.016-12.238). Patients with early latent syphilis were less likely to achieve serological response compared with those with primary or secondary syphilis (AOR, 0.317; 95% CI, 0.142-0.708). However, serological response was not affected by presence of spirochetemia or antibiotic regimens. PMID:24350785

Wu, B-R; Tsai, M-S; Yang, C-J; Sun, H-Y; Liu, W-C; Yang, S-P; Wu, P-Y; Su, Y-C; Chang, S-Y; Hung, C-C

2014-08-01

167

A possible mechanism of the nucleus accumbens and ventral pallidum 5-HT1B receptors underlying the antidepressant action of ketamine: a PET study with macaques  

PubMed Central

Ketamine is a unique anesthetic reagent known to produce various psychotic symptoms. Ketamine has recently been reported to elicit a long-lasting antidepressant effect in patients with major depression. Although recent studies provide insight into the molecular mechanisms of the effects of ketamine, the antidepressant mechanism has not been fully elucidated. To understand the involvement of the brain serotonergic system in the actions of ketamine, we performed a positron emission tomography (PET) study on non-human primates. Four rhesus monkeys underwent PET studies with two serotonin (5-HT)-related PET radioligands, [11C]AZ10419369 and [11C]DASB, which are highly selective for the 5-HT1B receptor and serotonin transporter (SERT), respectively. Voxel-based analysis using standardized brain images revealed that ketamine administration significantly increased 5-HT1B receptor binding in the nucleus accumbens and ventral pallidum, whereas it significantly reduced SERT binding in these brain regions. Fenfluramine, a 5-HT releaser, significantly decreased 5-HT1B receptor binding, but no additional effect was observed when it was administered with ketamine. Furthermore, pretreatment with 2,3-dihydroxy-6-nitro-7-sulfamoylbenzo(f)quinoxaline (NBQX), a potent antagonist of the glutamate ?-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) receptor, blocked the action of ketamine on the 5-HT1B receptor but not SERT binding. This indicates the involvement of AMPA receptor activation in ketamine-induced alterations of 5-HT1B receptor binding. Because NBQX is known to block the antidepressant effect of ketamine in rodents, alterations in the serotonergic neurotransmission, particularly upregulation of postsynaptic 5-HT1B receptors in the nucleus accumbens and ventral pallidum may be critically involved in the antidepressant action of ketamine. PMID:24399045

Yamanaka, H; Yokoyama, C; Mizuma, H; Kurai, S; Finnema, S J; Halldin, C; Doi, H; Onoe, H

2014-01-01

168

Biobarcodes: Antibodies and Nanosensors  

NSDL National Science Digital Library

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.

Network, Nanoscale I.; Industry, Oregon M.

2014-06-04

169

Antibodies to cholesterol.  

PubMed

Cholesterol-dependent complement activation has been proposed as a factor that might influence the pathogenesis of atherosclerosis. Although antibodies to cholesterol conjugates have been reported, cholesterol is widely regarded as a poorly immunogenic substance. Monoclonal IgM complement-fixing antibodies to cholesterol were obtained in the present study after immunizing mice with liposomes containing high amounts of cholesterol (71 mol % relative to phosphatidylcholine) and lipid A as an adjuvant. Clones were selected for the ability of secreted antibodies to react with liposomes containing 71% cholesterol but not with liposomes containing 43% cholesterol. The antibodies also reacted with crystalline cholesterol in a solid-phase enzyme-linked immunosorbent assay. Binding of monoclonal antibodies to the surface of crystalline cholesterol was demonstrated by electron microscopy by utilizing a second antibody (anti-IgM) labeled with colloidal gold. The immunization period required to induce monoclonal antibodies was very short (3 days) and a high fraction of the hybrid cells (at least 70%) were secreting detectable antibodies to cholesterol. The results demonstrate that cholesterol can be a highly immunogenic molecule and that complement-fixing antibodies to cholesterol can be readily obtained. PMID:3162316

Swartz, G M; Gentry, M K; Amende, L M; Blanchette-Mackie, E J; Alving, C R

1988-03-01

170

Expression of Recombinant Antibodies  

PubMed Central

Recombinant antibodies are highly specific detection probes in research, diagnostics, and have emerged over the last two decades as the fastest growing class of therapeutic proteins. Antibody generation has been dramatically accelerated by in vitro selection systems, particularly phage display. An increasing variety of recombinant production systems have been developed, ranging from Gram-negative and positive bacteria, yeasts and filamentous fungi, insect cell lines, mammalian cells to transgenic plants and animals. Currently, almost all therapeutic antibodies are still produced in mammalian cell lines in order to reduce the risk of immunogenicity due to altered, non-human glycosylation patterns. However, recent developments of glycosylation-engineered yeast, insect cell lines, and transgenic plants are promising to obtain antibodies with “human-like” post-translational modifications. Furthermore, smaller antibody fragments including bispecific antibodies without any glycosylation are successfully produced in bacteria and have advanced to clinical testing. The first therapeutic antibody products from a non-mammalian source can be expected in coming next years. In this review, we focus on current antibody production systems including their usability for different applications. PMID:23908655

Frenzel, Andre; Hust, Michael; Schirrmann, Thomas

2013-01-01

171

GE Healthcare Antibody Purification  

E-print Network

GE Healthcare Antibody Purification Handbook GE Healthcare imagination at work agination at work Separation Media Methodology and Applications 18-1115-69 Ion Exchange Chromatography & Chromatofocusing from GE Healthcare #12;Antibody Purification Handbook #12; Handbook 18-1037-46 AD Contents Introduction

Lebendiker, Mario

172

Antibodies with infinite affinity  

PubMed Central

Here we report an approach to the design and production of antibody/ligand pairs, to achieve functional affinity far greater than avidin/biotin. Using fundamental chemical principles, we have developed antibody/ligand pairs that retain the binding specificity of the antibody, but do not dissociate. Choosing a structurally characterized antibody/ligand pair as an example, we engineered complementary reactive groups in the antibody binding pocket and the ligand, so that they would be in close proximity in the antibody/ligand complex. Cross-reactions with other molecules in the medium are averted because of the low reactivity of these groups; however, in the antibody/ligand complex the effective local concentrations of the complementary reactive groups are very large, allowing a covalent reaction to link the two together. By eliminating the dissociation of the ligand from the antibody, we have made the affinity functionally infinite. This chemical manipulation of affinity is applicable to other biological binding pairs. PMID:11447282

Chmura, Albert J.; Orton, Molly S.; Meares, Claude F.

2001-01-01

173

Production Of Human Antibodies  

NASA Technical Reports Server (NTRS)

Process for making human monoclonal antibodies based on combination of techniques. Antibodies made active against specific antigen. Process involves in vivo immunization of human B lymphocyte cells in mice. B cells of interest enriched in vitro before fusion. Method potentially applicable to any antigen. Does not rely on use of Epstein-Barr virus at any step. Human lymphocytes taken from any source.

Sammons, David W.; Neil, Garry A.

1993-01-01

174

Therapeutic Recombinant Monoclonal Antibodies  

ERIC Educational Resources Information Center

During the last two decades, the rapid growth of biotechnology-derived techniques has led to a myriad of therapeutic recombinant monoclonal antibodies with significant clinical benefits. Recombinant monoclonal antibodies can be obtained from a number of natural sources such as animal cell cultures using recombinant DNA engineering. In contrast to…

Bakhtiar, Ray

2012-01-01

175

[Humanized antibodies as therapeutics].  

PubMed

Since 1997, nine humanized antibodies received the approval of the FDA to be used as drugs for the treatment of various diseases including transplant rejections, metastatic breast and colon cancers, leukaemia, non-Hodgkin lymphomas, allergic conditions or multiple sclerosis. This review describes techniques used to engineer these antibodies and presents the recent evolutions of these techniques : SDRs grafting or < abbreviated > CDRs grafting. Based on the illustrative examples of several antibodies, Mylotarg, Herceptin or Xolair, the therapeutic effectiveness of humanized antibodies are underlined and, with the example of Tysabri, the sometimes dramatic adverse effects associated with their clinical use is stressed. In a second part, this review presents some future and realistic avenues to improve the effectiveness of the humanized antibodies, to decrease their immunogenicity and to reduce their cost. PMID:16324646

Bellet, Dominique; Dangles-Marie, Virginie

2005-12-01

176

[Recombinant antibodies against bioweapons].  

PubMed

The threat posed by bioweapons (BW) could lead to the re-emergence of such deadly diseases as plague or smallpox, now eradicated from industrialized countries. The development of recombinant antibodies allows tackling this risk because these recombinant molecules are generally well tolerated in human medicine, may be utilized for prophylaxis and treatment, and because antibodies neutralize many BW. Recombinant antibodies neutralizing the lethal toxin of anthrax, botulinum toxins and the smallpox virus have in particular been isolated recently, with different technologies. Our approach, which uses phage-displayed immune libraries built from non-human primates (M. fascicularis) to obtain recombinant antibodies, which may later be super-humanized (germlinized), has allowed us to obtain such BWs-neutralizing antibodies. PMID:20035695

Thullier, Philippe; Pelat, Thibaut; Vidal, Dominique

2009-12-01

177

Antibodies for biodefense  

PubMed Central

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

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

2011-01-01

178

Antibody engineering for cancer therapy  

E-print Network

Antibodies targeting various tumor-associated antigens have been developed successfully to treat cancer. In this Thesis, novel antibodies and antibody-conjugate against two tumor antigens, AF-20 antigen and human aspartyl ...

Yeung, Yik Andy

2005-01-01

179

Antibody discovery: sourcing of monoclonal antibody variable domains.  

PubMed

Historically, antibody variable domains for therapeutic antibodies have been sourced primarily from the mouse IgG repertoire, and typically either chimerized or humanized. More recently, human antibodies from transgenic mice producing human IgG, phage display libraries, and directly from human B lymphocytes have been used more broadly as sources of antibody variable domains for therapeutic antibodies. Of the total 36 antibodies approved by major maket regulatory agencies, the variable domain sequences of 26 originate from the mouse. Of these, four are marketed as murine antibodies (of which one is a mouse-rat hybrid IgG antibody), six are mouse-human chimeric antibodies, and 16 are humanized. Ten marketed antibodies have originated from human antibody genes, three isolated from phage libraries of human antibody genes and seven from transgenic mice producing human antibodies. Five antibodies currently in clinical trials have been sourced from camelids, as well as two from non-human primates, one from rat, and one from rabbit. Additional sources of antibody variable domains that may soon find their way into the clinic are potential antibodies from sharks and chickens. Finally, the various methods for retrieval of antibodies from humans, mouse and other sources, including various display technologies and amplification directly from B cells, are described. PMID:24168292

Strohl, William R

2014-03-01

180

Monoclonal antibody "gold rush".  

PubMed

The market, sales and regulatory approval of new human medicines, during the past few years, indicates increasing number and share of new biologics and emergence of new multibillion dollar molecules. The global sale of monoclonal antibodies in 2006 were $20.6 billion. Remicade had annual sales gain of $1 billion during the past 3 years and five brands had similar increase in 2006. Rituxan with 2006 sales of $4.7 billion was the best selling monoclonal antibody and biological product and the 6th among the top selling medicinal brand. It may be the first biologic and monoclonal antibody to reach $10 billion annual sales in the near future. The strong demand from cancer and arthritis patients has surpassed almost all commercial market research reports and sales forecast. Seven monoclonal antibody brands in 2006 had sales exceeding $1 billion. Humanized or fully human monoclonal antibodies with low immunogenicity, enhanced antigen binding and reduced cellular toxicity provide better clinical efficacy. The higher technical and clinical success rate, overcoming of technical hurdles in large scale manufacturing, low cost of market entry and IND filing, use of fully human and humanized monoclonal antibodies has attracted funds and resources towards R&D. Review of industry research pipeline and sales data during the past 3 years indicate a real paradigm shift in industrial R&D from pharmaceutical to biologics and monoclonal antibodies. The antibody bandwagon has been joined by 200 companies with hundreds of new projects and targets and has attracted billions of dollars in R&D investment, acquisitions and licensing deals leading to the current Monoclonal Antibody Gold Rush. PMID:17691940

Maggon, Krishan

2007-01-01

181

Heart antibodies in cardiomyopathies.  

PubMed Central

The reported frequency of circulating heart reactive antibodies in cardiomyopathies has varied and their significance is unknown. In this study such antibodies were sought in patients with primary congestive and hypertrophic cardiomyopathies and other heart diseases. Standard "single sandwich" and the more sensitive "double sandwich" indirect immunofluorescence techniques failed to disclose a significant difference between any cardiomyopathic group and controls in repeated experiments. With both techniques results were subject to considerable method-specific artefacts and observer variation. No published work associating heart antibodies detected by immunofluorescence methods with cariomyopathies adequately takes these into account. PMID:7028058

Trueman, T; Thompson, R A; Cummins, P; Littler, W A

1981-01-01

182

Mining human antibody repertoires  

PubMed Central

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 naďve, 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

2010-01-01

183

Intracellular antibody immunity.  

PubMed

Antibodies allow the immune system to target pathogens despite their tremendous diversity and rapid evolution. Once bound to a pathogen, antibodies induce a broad range of effector mechanisms, including phagocytosis and complement. However, these mechanisms are all initiated in the extracellular space, meaning that pathogens like viruses evade them upon infection of their target cells. Recently, it has been shown that, in addition to mediating extracellular immune responses, antibodies also activate immunity inside infected cells. Antibodies that are bound to the surface of non-enveloped viruses or bacteria are carried into the cell during pathogen entry. Once inside the cell, these pathogen-attached antibodies are recognised by a highly conserved, high affinity cytosolic antibody receptor called TRIM21. TRIM21 initiates both sensor and effector responses that reduce viral replication and induce an antiviral state. These responses are an important part of antiviral immunity and the removal of TRIM21 results in uncontrolled viraemia and death in a mouse model of infection. PMID:24722852

Watkinson, Ruth E; McEwan, William A; James, Leo C

2014-07-01

184

STUDIES ON ANTIBODY PRODUCTION  

PubMed Central

A method for the specific histochemical demonstration of antibody in cells and parts of cells is described. It consists of carrying out a two stage immunological reaction on frozen sections of tissues: (a) allowing reaction between antibody in the tissue and dilute antigen applied in vitro, and (b) the detection of those areas where this antigen has been specifically absorbed by means of a precipitin reaction carried out with fluorescein-labelled antibody. Examination under the fluorescence microscope reveals the yellow-green fluorescence of fluorescein over those areas where a precipitate has formed. A study of the hyperimmune rabbit on the first few days after the last of a series of intravenous antigen injections reveals that antibody against human ?-globulin or ovalbumin is present in groups of plasma cells in the red pulp of the spleen, the medullary areas of lymph nodes, the submucosa of the ileum, and the portal connective tissue of the liver. Because of extensive non-specific reactions, the bone marrow could not be examined. Small amounts of antibody were occasionally visible in cells in the lymphoid follicles of the spleen and lymph nodes, so that a minor contribution by lymphocytes to antibody synthesis cannot be excluded. PMID:14392240

Coons, Albert H.; Leduc, Elizabeth H.; Connolly, Jeanne M.

1955-01-01

185

Antibody-targeted vaccines.  

PubMed

The specificity and high affinity binding of antibodies provides these molecules with ideal properties for delivering a payload to target cells. This concept has been commercialized for cancer therapies using toxin- or radionucleotide-conjugated antibodies that are designed to selectively deliver cytotoxic molecules to cancer cells. Exploiting the same effective characteristics of antibodies, antibody-targeted vaccines (ATV) are designed to deliver disease-specific antigens to professional antigen-presenting cells (APCs), thus enabling the host's immune system to recognize and eliminate malignant or infected cells through adaptive immunity. The concept of ATVs has been in development for many years, and recently has entered clinical trials. Early studies with ATVs focused on the ability to induce humoral immunity in the absence of adjuvants. More recently, ATVs targeted to C-type lectin receptors have been exploited for induction of potent helper and cytolytic T-cell responses. To maximize their stimulatory capacity, the ATVs are being evaluated with a variety of adjuvants or other immunostimulatory agents. In the absence of co-administered immunostimulatory signals, APC-targeting can induce antigen-specific tolerance and, thus, may also be exploited in developing specific treatments for autoimmune and allergic diseases, or for preventing transplant rejection. The successful clinical application of this new class of antibody-based products will clearly depend on using appropriate combinations with other strategies that influence the immune system. PMID:17530028

Keler, T; He, L; Ramakrishna, V; Champion, B

2007-05-28

186

The Transition from Closed to Open Conformation of Treponema pallidum Outer Membrane-associated Lipoprotein TP0453 Involves Membrane Sensing and Integration by Two Amphipathic Helices*  

PubMed Central

The molecular architecture and composition of the outer membrane (OM) of Treponema pallidum (Tp), the noncultivable agent of venereal syphilis, differ considerably from those of typical Gram-negative bacteria. Several years ago we described TP0453, the only lipoprotein associated with the inner leaflet of the Tp OM. Whereas polypeptides of other treponemal lipoproteins are hydrophilic, non-lipidated TP0453 can integrate into membranes, a property attributed to its multiple amphipathic helices (AHs). Furthermore, membrane integration of the TP0453 polypeptide was found to increase membrane permeability, suggesting the molecule functions in a porin-like manner. To better understand the mechanism of membrane integration of TP0453 and its physiological role in Tp OM biogenesis, we solved its crystal structure and used mutagenesis to identify membrane insertion elements. The crystal structure of TP0453 consists of an ?/?/?-fold and includes five stably folded AHs. In high concentrations of detergent, TP0453 transitions from a closed to open conformation by lateral movement of two groups of AHs, exposing a large hydrophobic cavity. Triton X-114 phase partitioning, liposome floatation assay, and bis-1-anilino-8-naphthalenesulfonate binding revealed that two adjacent AHs are critical for membrane sensing/integration. Using terbium-dipicolinic acid complex-loaded large unilamellar vesicles, we found that TP0453 increased efflux of fluorophore only at acidic pH. Gel filtration and cross-linking experiments demonstrated that one AH critical for membrane sensing/insertion also forms a dimeric interface. Based on structural dynamics and comparison with Mycobacterium tuberculosis lipoproteins LprG and LppX, we propose that TP0453 functions as a carrier of lipids, glycolipids, and/or derivatives during OM biogenesis. PMID:21965687

Luthra, Amit; Zhu, Guangyu; Desrosiers, Daniel C.; Eggers, Christian H.; Mulay, Vishwaroop; Anand, Arvind; McArthur, Fiona A.; Romano, Fabian B.; Caimano, Melissa J.; Heuck, Alejandro P.; Malkowski, Michael G.; Radolf, Justin D.

2011-01-01

187

The Crystal Structure of Zn(II)-Free Treponema pallidum TroA, a Periplasmic Metal-Binding Protein, Reveals a Closed Conformation  

PubMed Central

We previously demonstrated that Treponema pallidum TroA is a periplasmic metal-binding protein (MBP) with a distinctive alpha-helical backbone. To better understand the mechanisms of metal binding and release by TroA, we determined the crystal structure of the apoprotein at a resolution of 2.5 Ĺ and compared it to that of the Zn(II)-bound form (Protein Data Bank accession code 1toa). apo-TroA shows a conformation even more closed than that of its Zn(II)-bound counterpart due to a 4° tilt of the C-terminal domain (residues 190 through 308) about an axis parallel to the poorly flexible backbone helix. This domain tilting pushes two loops (residues 248 through 253 and 277 through 286) towards the metal-binding site by more than 1 Ĺ, resulting in an unfavorable interaction of I251 with D66. To avoid this contact, D66 shifts towards H68, one of the four Zn(II)-coordinating residues. The approach of this negative charge coincides with the flipping of the imidazole side chain of H68, resulting in the formation of a new hydrogen bond. The conformational change of H68, along with a slight rearrangement of D279, a C-terminal domain Zn(II)-coordinating residue, distorts the metal-binding site geometry, presumably causing the release of the bound metal ion. Ligand binding and release by TroA, and presumably by other members of the MBP cluster, differs from the “Venus flytrap” mechanism utilized by bacterial nonmetal solute-binding receptors. PMID:11914363

Lee, Yong-Hwan; Dorwart, Michael R.; Hazlett, Karsten R. O.; Deka, Ranjit K.; Norgard, Michael V.; Radolf, Justin D.; Hasemann, Charles A.

2002-01-01

188

HIV-1, HBV, HCV, HTLV, HPV-16/18, and Treponema pallidum Infections in a Sample of Brazilian Men Who Have Sex with Men  

PubMed Central

Background Men who have sex with men (MSM) are more vulnerable to blood-borne infections and/or sexually-transmitted infections (STI). This study was conducted to estimate the prevalences of mono and co-infections of HIV-1 and other blood-borne/STIs in a sample of MSM in Campinas, Brazil. Methods Responding Driven Sampling (RDS) was used for recruitment of MSM. Serum samples collected from 558 MSM were analyzed for the presence of serological markers for HIV-1, HBV, HCV, HTLV, HPV-16/18, and T. pallidum infections. Results The highest prevalences of infection in serum samples were found for HPV-16 and 18 (31.9% and 20.3%, respectively). Approximately 8% of the study population showed infection with HIV-1, and within that group, 27.5% had recently become infected with HIV-1. HBV infection and syphilis were detected in 11.4% and 10% of the study population, respectively, and the rates of HTLV and HCV infection were 1.5% and 1%, respectively. With the exception of HTLV, all other studied infections were usually found as co-infections rather then mono-infections. The rates of co-infection for HCV, HPV-18, and HIV-1 were the highest among the studied infections (100%, 83%, and 85%, respectively). Interestingly, HTLV infection was usually found as a mono-infection in the study group, whereas HCV was found only as a co-infection. Conclusions The present findings highlight the need to educate the MSM population concerning their risk for STIs infections and methods of prevention. Campaigns to encourage vaccination against HBV and HPV could decrease the rates of these infections in MSM. PMID:25083768

Soares, Caroline C.; Georg, Ingebourg; Lampe, Elisabeth; Lewis, Lia; Morgado, Mariza G.; Nicol, Alcina F.; Pinho, Adriana A.; Salles, Regina C. S.; Teixeira, Sylvia L. M.; Vicente, Ana Carolina P.; Viscidi, Raphael P.; Gomes, Selma A.

2014-01-01

189

Glycoproteomic Analysis of Antibodies*  

PubMed Central

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

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

2013-01-01

190

An Antibody Molecule  

NSDL National Science Digital Library

An antibody molecule. (A) Schematic drawing of a typical antibody molecule. As indicated, this protein is Y-shaped and has two identical binding sites for its antigen, one on either arm of the 3Y.2 The protein is composed of four polypeptide chains (two identical heavy chains and two identical and smaller light chains) held together by disulfide bonds. Each chain is made up of several different domains, here shaded either blue or gray. The antigen-binding site is formed where a heavy chain variable domain (VH) and a light chain variable domain (VL) come close together. These are the domains that differ most in their sequence and structure in different antibodies. (B) Ribbon drawing of a light chain showing the parts of the VL domain most closely involved in binding to the antigen in red; these contribute half of the fingerlike loops that fold around each of the antigen molecules in (A).

BEGIN:VCARD VERSION:2.1 FN:Martin Raff N:Raff;Martin REV:2005-04-15 END:VCARD; BEGIN:VCARD VERSION:2.1 FN:Julian Lewis N:Lewis;Julian REV:2005-04-15 END:VCARD; BEGIN:VCARD VERSION:2.1 FN:Alexander Johnson N:Johnson;Alexander REV:2005-04-15 END:VCARD; BEGIN:VCARD VERSION:2.1 FN:Dennis Bray N:Bray;Dennis REV:2005-04-15 END:VCARD; BEGIN:VCARD VERSION:2.1 FN:Bruce Alberts N:Alberts;Bruce REV:2005-04-15 END:VCARD; BEGIN:VCARD VERSION:2.1 FN:Keith Roberts N:Roberts;Keith REV:2005-04-15 END:VCARD

1998-07-01

191

Antibody-Antigen Interactions  

NSDL National Science Digital Library

The experimental protocol in this Web site is just one of many microbiology resources provided by the University of Leicester. The procedure guides students in finding the antibody concentration of a test antiserum and the number of antibody binding sites on an antigen molecule. A results graph and correct answers to the required calculations are given, providing the option of performing a virtual experiment in lieu of an actual one. This activity is probably most appropriate for high school and undergraduate level biology labs.

Cann, Alan.

2010-01-05

192

Demonstration of specific 19S(IgM) antibodies in untreated and treated syphilis. Comparative studies of the 19S(IgM)-FTA test, the 19S(IgM)-TPHA test, and the solid phase haemadsorption assay.  

PubMed Central

Sera from 408 patients with untreated or treated syphilis were examined by three different tests to demonstrate treponema-specific 19S(IgM) antibodies. Antibody titers in the 19S(IgM)-fluorescent treponemal antibody (FTA) test and the solid phase haemadsorption assay (SPHA) did not correlate. The 19S(IGM)-Treponema pallidum haemagglutination assay (TPHA) and the SPHA partly correlated based on the concentration of treponema-specific 19S(IgM) antibodies in the patient's serum. Under experimental conditions antibody titres in the 19S(IGM)-FTA test and the 19S(IgM)-TPHA correlated consistently. For specificity and sensitivity, the 19S(IgM)-FTA test correlated best with the clinical findings in both untreated and successfully treated patients. Although the 19S(IgM)-TPHA has about the same degree of specificity, the reading of the results is technically more complicated. The specificity of the SPHA was very high. In patients with untreated syphilis, however, the SPHA is adversely affected by a high rate of false non-reactive results, since it consists of two reactions with appreciable differences in sensitivity. Thus, higher sensitivity cannot be expected in the SPHA. An immunoadsorption technique using an adequate antigen and a specific, enzyme-labelled antiserum might provide an alternative test which is simple to perform, highly specific, and consistently sensitive. Images PMID:7034857

Muller, F; Lindenschmidt, E G

1982-01-01

193

Therapeutic antibody engineering  

PubMed Central

It is an important event in any knowledge area when an authority in the field decides that it is time to share all accumulated knowledge and learnings by writing a text book. This does not occur often in the biopharmaceutical industry, likely due to both the highly dynamic environment with tight timelines and policies and procedures at many pharmaceutical companies that hamper knowledge sharing. To take on a task like this successfully, a strong drive combined with a desire and talent to teach, but also an accommodating and stimulating environment is required. Luckily for those interested in therapeutic monoclonal antibodies, Dr. William R. Strohl decided about two years ago that the time was right to write a book about the past, present and future of these fascinating molecules. Dr. Strohl’s great expertise and passion for biotechnology is evident from his life story and his strong academic and industry track record. Dr. Strohl pioneered natural product biotechnology, first in academia as a full professor of microbiology and biochemistry at Ohio State University in Columbus, Ohio and later in industry while at Merck. Despite his notable advances in recombinant natural products, industry interest in this area waned and in 2001 Dr. Strohl sought new opportunities by entering the field of antibody therapeutics. He initiated antibody discovery through phage display at Merck, and then moved to Centocor Research and Development Inc. (now Janssen Biotech, Inc.) in 2008 to head Biologics Research, where he now directs the discovery of innovative therapeutic antibody candidates.

Parren, Paul W.H.I.; Lugovskoy, Alexey A.

2013-01-01

194

Polyclonal antibody catalytic variability.  

PubMed Central

We have performed a systematic variability study of polyclonal antibody catalysis by using five rabbits immunized with the same hapten. Important results from this work are the following. (1) Similarities were observed in the catalytic polyclonal antibodies derived from all five rabbits. Four of the five rabbits produced polyclonal samples that were nearly the same in terms of catalytic activity, whereas the fifth rabbit, designated as rabbit 2, displayed a somewhat higher level of catalytic activity. The catalytic activities (as kcat/kuncat) of these polyclonal samples were similar to that from the best murine monoclonal antibody that had been previously elicited by the same hapten. (2) Titre was not an accurate indicator of polyclonal antibody catalytic activity. (3) A mathematical analysis to describe a distribution of Michaelis-Menten catalysts was performed to help interpret our results. (4) Kinetic analysis indicated that the binding parameters of the different samples were remarkably homogeneous, because one or two components were all that were required to fit the on-rate and off-rate data satisfactorily. Interestingly, the most active catalytic polyclonal sample, that from rabbit 2, displayed the slowest off-rate (so slow it could not be measured) and thus the highest overall affinity. (5) Catalytic analysis of eluted fractions of antibody from a substrate column indicated that each polyclonal sample was also relatively homogeneous in terms of catalytic parameters. The main conclusion of our study is that for this hapten-animal system, the overall catalytic immune response is relatively consistent at two levels. Consistent catalytic activity was observed between the polyclonal samples elicited in the different animals, and the elicited hapten-specific polyclonal antibodies were relatively homogeneous in terms of binding and catalytic parameters within each immunized animal. The observed similarities of the catalytic activity in the different animals is surprising, because the immune response is based on specific binding of antibodies to hapten. There is no known selective pressure to maintain consistent levels of catalytic activity. Our results can therefore be interpreted as providing evidence that for this hapten there is a fixed relationship between hapten structure and catalytic activity and/or consistent genetic factors that dominate the catalytic immune response. PMID:9576860

Stephens, D B; Thomas, R E; Stanton, J F; Iverson, B L

1998-01-01

195

Humanized Antibodies for Antiviral Therapy  

NASA Astrophysics Data System (ADS)

Antibody therapy holds great promise for the treatment of cancer, autoimmune disorders, and viral infections. Murine monoclonal antibodies are relatively easy to produce but are severely restricted for therapeutic use by their immunogenicity in humans. Production of human monoclonal antibodies has been problematic. Humanized antibodies can be generated by introducing the six hypervariable regions from the heavy and light chains of a murine antibody into a human framework sequence and combining it with human constant regions. We humanized, with the aid of computer modeling, two murine monoclonal antibodies against herpes simplex virus gB and gD glycoproteins. The binding, virus neutralization, and cell protection results all indicate that both humanized antibodies have retained the binding activities and the biological properties of the murine monoclonal antibodies.

Co, Man Sung; Deschamps, Marguerite; Whitley, Richard J.; Queen, Cary

1991-04-01

196

Anti-insulin antibody test  

MedlinePLUS

Insulin antibodies - serum; Insulin Ab test ... Normally, there are no antibodies against insulin in your blood. Normal value ranges may vary slightly among different laboratories. Some labs use different measurements or ...

197

The Art of Making Antibodies.  

ERIC Educational Resources Information Center

Provides background information for teachers on the nature and production of antibodies. Points out that the production of monoclonal antibodies blends the malignant with the beneficial to create a medical tool of exciting potential. (JN)

Headon, Denis R.

1986-01-01

198

Human Antibodies to Bovine ?-Globulin  

Microsoft Academic Search

Antibodies to bovine ?-globulin (anti-BGG antibodies) were detectable by a radio-immunoassay in 70% of healthy blood donors but, generally, the titres were low. Significantly increased concentrations of anti-BGG antibodies were found in patients lacking IgA but not in patients with allergic disorders. The anti-BGG antibodies were shown to give rise to falsely high IgE values in the radio-immunosorbent test for

T. Foucard; H. Bennich; S. G. O. Johansson; U. Lundkvist

1975-01-01

199

Antibody-gold cluster conjugates  

DOEpatents

Antibody- or antibody fragment-gold cluster conjugates are shown wherein the conjugate size can be about 5.0 nm. Methods and reagents are disclosed in which antibodies or Fab' fragments thereof are covalently bound to a stable cluster of gold atoms. 2 figs.

Hainfeld, J.F.

1988-06-28

200

Her2 Monoclonal Antibodies, Antibody Drug Conjugates, and Site Specific Antibody Conjugate Methods  

Cancer.gov

Antibody drug conjugates (ADC) can demonstrate high efficacy as cancer therapeutics, however, much more can be done to improve their efficacy and safety profile. Site-specific antibody drug conjugation is a promising way to do this.

201

Antibody Engineering and Therapeutics Conference  

PubMed Central

The Antibody Engineering and Therapeutics conference, which serves as the annual meeting of The Antibody Society, will be held in Huntington Beach, CA from Sunday December 8 through Thursday December 12, 2013. The scientific program will cover the full spectrum of challenges in antibody research and development, and provide updates on recent progress in areas from basic science through approval of antibody therapeutics. Keynote presentations will be given by Leroy Hood (Institute of System Biology), who will discuss a systems approach for studying disease that is enabled by emerging technology; Douglas Lauffenburger (Massachusetts Institute of Technology), who will discuss systems analysis of cell communication network dynamics for therapeutic biologics design; David Baker (University of Washington), who will describe computer-based design of smart protein therapeutics; and William Schief (The Scripps Research Institute), who will discuss epitope-focused immunogen design.   In this preview of the conference, the workshop and session chairs share their thoughts on what conference participants may learn in sessions on: (1) three-dimensional structure antibody modeling; (2) identifying clonal lineages from next-generation data sets of expressed VH gene sequences; (3) antibodies in cardiometabolic medicine; (4) the effects of antibody gene variation and usage on the antibody response; (5) directed evolution; (6) antibody pharmacokinetics, distribution and off-target toxicity; (7) use of knowledge-based design to guide development of complementarity-determining regions and epitopes to engineer or elicit the desired antibody; (8) optimizing antibody formats for immunotherapy; (9) antibodies in a complex environment; (10) polyclonal, oligoclonal and bispecific antibodies; (11) antibodies to watch in 2014; and (12) polyreactive antibodies and polyspecificity.

Almagro, Juan Carlos; Gilliland, Gary L; Scott, Jamie; Larrick, James W; Pluckthun, Andreas; Veldman, Trudi; Adams, Gregory P; Parren, Paul WHI; Chester, Kerry A; Bradbury, Andrew; Reichert, Janice M; Huston, James S

2013-01-01

202

Infertility and Antiphospholipid Antibodies  

Microsoft Academic Search

Whether antiphospholipid antibodies (aPL) play a pathogenic role in infertility is highly controversial. aPL have been suggested to represent one potential etiology of infertility, specifically in patients with unexplained implantation failure following in vitro fertilization (IVF). The rationale is appealing, as it represents a logical extension of the demonstrated pathogenicity of aPL in contributing to recurrent spontaneous abortion, where mechanisms

Lisa R. Sammaritano

203

Commercial antibodies and their validation  

PubMed Central

Despite an impressive growth in the business of research antibodies a general lack of trust in commercial antibodies remains in place. A variety of issues, each one potentially causing an antibody to fail, underpin the frustrations that scientists endure. Lots of money goes to waste in buying and trying one failing antibody after the other without realizing all the pitfalls that come with the product: Antibodies can get inactivated, both the biological material and the assay itself can potentially be flawed, a single antibody featuring in many different catalogues can be deemed as a set of different products, and a bad choice of antibody type, wrong dilutions, and lack of proper validation can all jeopardize the intended experiments. Antibodies endorsed by scientific research papers do not always meet the scientist’s requirements either due to flawed specifications, or due to batch-to-batch variations. Antibodies can be found with Quality Control data obtained from previous batches that no longer represent the batch on sale. In addition, one cannot assume that every antibody is fit for every application. The best chance of success is to try an antibody that already was confirmed to perform correctly in the required platform. PMID:25324967

Voskuil, JLA

2014-01-01

204

Natural antibodies and cancer.  

PubMed

Immunity is not only responsible for recognition and elimination of infectious particles, but also for removal of cellular waste, modified self structures and transformed cells. Innate or natural immunity acts as a first line defense and is also the link to acquired immunity and memory. A striking phenomenon of immunity against malignant cells is that neither in animals nor in humans affinity-maturated tumor-specific IgG antibodies have been detected so far. All tumor-specific isolated antibodies were germ-line coded natural IgM antibodies. It's also a fact that these IgM's preferentially bind to carbohydrate epitopes on post-transcriptionally modified surface receptors and that they all induce a cancer-specific apoptosis, by triggering the intrinsic apoptotic pathway. From an evolutionary point of view, this makes sense because cancer cells are not infectious, so there is no need for memory. Natural IgMs bind to conservative structures because they are coded by a limited set of genes and they use apoptosis, the "clean" way of killing, to avoid inflammatory processes. PMID:17826951

Vollmers, H Peter; Brändlein, Stephanie

2007-12-01

205

Pneumocystis carinii antibody testing.  

PubMed Central

Sera from blood donors and patients from all over Scotland were examined by indirect immunofluorescence using Pneumocystis carinii antigen from infected rat lung. Antibody was found in 76 of 488 (15.6%) of patients tested on clinical grounds but in only 13 of 148 (8.8%) blood donors. The antibody rates were higher in disease groups likely to have or develop P carinii pneumonia: in those with histologically confirmed or strongly suspected P carinii pneumonia the rate was 14 of 24 (58.3%); in those who had undergone transplantation eight of 24 (33.3%); in those who were immunosuppressed five of 16 (31.2%); in those who were human immunodeficiency virus antibody (HIV) positive 11 of 43 (25.6%); in those with malignancy 34 of 233 (14.6%); and in those with chest infection 10 of 85 (11.7%). P carinii pneumonia was confirmed or likely in four of 45 (8.8%) patients with titres of 1/8-1/16 and in three of seven (42.8%) in those with titres of greater than or equal to 1/128. Seroconversion or rising titre was detected in seven of 13 (53.8%) cases of confirmed or likely P carinii pneumonia compared with 10 of 93 (10.7%) in other patients. Diagnosis of P carinii infection can therefore be assisted by positive immunofluorescence results, but negative serology does not exclude infection. PMID:2671053

Chatterton, J M; Joss, A W; Williams, H; Ho-Yen, D O

1989-01-01

206

How antibodies use complement to regulate antibody responses.  

PubMed

Antibodies, forming immune complexes with their specific antigen, can cause complete suppression or several 100-fold enhancement of the antibody response. Immune complexes containing IgG and IgM may activate complement and in such situations also complement components will be part of the immune complex. Here, we review experimental data on how antibodies via the complement system upregulate specific antibody responses. Current data suggest that murine IgG1, IgG2a, and IgG2b upregulate antibody responses primarily via Fc-receptors and not via complement. In contrast, IgM and IgG3 act via complement and require the presence of complement receptors 1 and 2 (CR1/2) expressed on both B cells and follicular dendritic cells. Complement plays a crucial role for antibody responses not only to antigen complexed to antibodies, but also to antigen administered alone. Lack of C1q, but not of Factor B or MBL, severely impairs antibody responses suggesting involvement of the classical pathway. In spite of this, normal antibody responses are found in mice lacking several activators of the classical pathway (complement activating natural IgM, serum amyloid P component (SAP), specific intracellular adhesion molecule-grabbing non-integrin R1 (SIGN-R1) or C-reactive protein. Possible explanations to these observations will be discussed. PMID:25001046

Sörman, Anna; Zhang, Lu; Ding, Zhoujie; Heyman, Birgitta

2014-10-01

207

The future of monoclonal antibody technology  

PubMed Central

With the rapid growth of monoclonal antibody-based products, new technologies have emerged for creating modified forms of antibodies, including fragments, conjugates and multi-specific antibodies. We created a database of 450 therapeutic antibodies in development to determine which technologies and indications will constitute the “next generation” of antibody products. We conclude that the antibodies of the future will closely resemble the antibodies that have already been approved for commercial sale. PMID:20676053

Zider, Alexander

2010-01-01

208

Monoclonal antibodies for treating cancer  

SciTech Connect

The purpose of this study is to assess the current status of in-vivo use of monoclonal antibodies for treating cancer. Publications appearing between 1980 and 1988 were identified by computer searches using MEDLINE and CANCERLIT, by reviewing the table of contents of recently published journals, and by searching bibliographies of identified books and articles. More than 700 articles, including peer-reviewed articles and book chapters, were identified and selected for analysis. The literature was reviewed and 235 articles were selected as relevant and representative of the current issues and future applications for in-vivo monoclonal antibodies for cancer therapy and of the toxicity and efficacy which has been associated with clinical trials. Approaches include using antibody alone (interacting with complement or effector cells or binding directly with certain cell receptors) and immunoconjugates (antibody coupled to radioisotopes, drugs, toxins, or other biologicals). Most experience has been with murine antibodies. Trials of antibody alone and radiolabeled antibodies have confirmed the feasibility of this approach and the in-vivo trafficking of antibodies to tumor cells. However, tumor cell heterogeneity, lack of cytotoxicity, and the development of human antimouse antibodies have limited clinical efficacy. Although the immunoconjugates are very promising, heterogeneity and the antimouse immune response have hampered this approach as has the additional challenge of chemically or genetically coupling antibody to cytotoxic agents. As a therapeutic modality, monoclonal antibodies are still promising but their general use will be delayed for several years. New approaches using human antibodies and reducing the human antiglobulin response should facilitate treatment. 235 references.

Dillman, R.O. (Hoag Cancer Center, Newport Beach, CA (USA))

1989-10-01

209

Monoclonal antibody therapy of cancer  

Microsoft Academic Search

The most significant recent advances in the application of monoclonal antibodies (mAbs) to oncology have been the introduction and approval of bevacizumab (Avastin), an anti–vascular endothelial growth factor antibody, and of cetuximab (Erbitux), an anti–epidermal growth factor antibody. In combination with standard chemotherapy regimens, bevacizumab significantly prolongs the survival of patients with metastatic cancers of the colorectum, breast and lung.

Gregory P Adams; Louis M Weiner

2005-01-01

210

Brucella antibodies in sudanese camels  

Microsoft Academic Search

Sera of 740 camels of both sexes from three regions of Sudan were tested for antibodies toBrucella abortus. The overall incidence of antibodies was 4.9%. The highest positive number of samples (7.5%) was from the Eastern Region followed by Darfur Region (3.1%) and the Central Region (2.0%).Brucella antibodies were as frequent in males (5.6%) as females (4.5%).

H. Abu Damir; S. J. Kenyon; Amna E. Khalaf Alla; O. F. Idris

1984-01-01

211

Evaluation of multiplex real-time PCR for detection of Haemophilus ducreyi, Treponema pallidum, herpes simplex virus type 1 and 2 in the diagnosis of genital ulcer disease in the Rakai District, Uganda  

PubMed Central

Objective: To develop a real-time PCR assay that reliably and accurately detects the predominant sexually transmitted aetiological agents of genital ulcer disease (GUD) (Haemophilus ducreyi, Treponema pallidum and herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2)) and to assess the use of real-time PCR diagnostic testing in a rural African field site. Methods: Two multiplex real-time PCR reactions were used to detect H ducreyi/and HSV-1/HSV-2 in ulcer swabs from 100 people with symptomatic genital ulcers in rural Rakai, Uganda. Results were compared with syphilis, HSV-1 and HSV-2 serology. Results: Of 100 GUD samples analysed from 43 HIV positive and 57 HIV negative individuals, 71% were positive for one or more sexually transmitted infection (STI) pathogens by real-time PCR (61% for HSV-2, 5% for T pallidum, 3% for HSV-1, 1% for H ducreyi and 1% for dual H ducreyi/HSV-2). The frequency of HSV in genital ulcers was 56% (32/57) in HIV negative individuals and 77% (33/43) in HIV positive individuals (p=0.037). Assay reproducibility was evaluated by repeat PCR testing in the USA with 96% agreement (?=0.85). Conclusions: STI pathogens were detected in the majority of GUD swab samples from symptomatic patients in Rakai, Uganda, by real-time PCR. HSV-2 was the predominant cause of genital ulcers. Real-time PCR technology can provide sensitive, rapid and reproducible evaluation of GUD aetiology in a resource-limited setting. PMID:19066198

Suntoke, T R; Hardick, A; Tobian, A A R; Mpoza, B; Laeyendecker, O; Serwadda, D; Opendi, P; Gaydos, C A; Gray, R H; Wawer, M J; Quinn, T C; Reynolds, S J

2009-01-01

212

Antiphospholipid antibodies and infertility.  

PubMed

Since the late 1980s some publications have proposed that antiphospholipid antibodies (aPL) may have some relationship with infertility, considering reported deleterious effects that aPL exert on trophoblast proliferation and growth. Although not included in current classification criteria for antiphospholipid syndrome, many physicians investigate for aPL in patients with a history of infertility, including antibodies not listed in classification criteria, and most of those patients will receive anticoagulant therapy if any of those antibodies have a result considered positive. A review of literature was conducted searching for studies that investigated the association of aPL and infertility and if aPL positivity alters in vitro fertilization (IVF) outcome. The definition of infertility, routine work-up to exclude other causes of infertility, definition of IVF failure as inclusion criteria and control populations were heterogeneous among studies. Most of them enrolled women over 40 years of age, and exclusion of other confounding factors was also inconsistent. Of 29 studies that assessed aPL positivity rates in infertile women, the majority had small sample sizes, implying a lack of power, and 13 (44.8%) reported higher frequency of aPL in infertile patients compared to controls, but most of them investigated a panel of non-criteria aPL tests, whose clinical significance is highly controversial. Only two studies investigated all three criteria tests, and medium-high titer of anticardiolipin cut-off conforming to international guidelines was used in one study. Considering IVF outcome, there was also disparity in this definition: few studies assessed the live birth rate, others the implantation rate. Of 14 publications that addressed the relationship between aPL and IVF outcome, only two described a detrimental effect of these autoantibodies. In conclusion, available data do not support an association between aPL and infertility, and aPL positivity does not seem to influence IVF outcome. Well-designed clinical studies recruiting women with a clear diagnosis of infertility and a high-risk aPL profile should be performed to test whether clinically relevant aPL do-or not-exert an effect on human fertility. PMID:25228713

Chighizola, Cb; de Jesus, Gr

2014-10-01

213

Micromechanical antibody sensor  

DOEpatents

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.

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

214

Natural antibody - Biochemistry and functions  

PubMed Central

Natural antibodies have been common knowledge in the scientific community for more than half a century. Initially disregarded, their functions have garnered a newfound interest recently. Natural antibodies are usually polyreactive IgM antibodies and are implicated in numerous physiologic and pathologic processes. Current research demonstrates they play a role in adaptive and innate immune responses, autoimmunity, and apoptosis. Evidence exists that they are involved in the modulation of neurodegenerative disorders and malignancy. Furthermore, natural antibodies have been implicated in ischemia reperfusion injury and atherosclerosis. As such the study of natural antibodies may provide new insight into normal physiologic processes whilst concurrently paving the road for a wide-range of possible therapeutic options.

Rahyab, Ali Seyar; Alam, Amit; Kapoor, Aricka; Zhang, Ming

2012-01-01

215

Bio-Rad Laboratories I n f e c t I o u s D I s e a s e c o n t r o l s Syphilis Total  

E-print Network

Immunoglobulin G and Immunoglobulin M antibodies to Treponema pallidum and non-treponemal antibodies (Reagin). 60 pallidum IgG Treponema pallidum IgM Reagin Refer to the package insert for performance and stability claims Representative for more information. Discover the power of UnityTM Solutions at www.QCNet.com. Analytes Treponema

Rodriguez, Carlos

216

Computer-aided antibody design  

PubMed Central

Recent clinical trials using antibodies with low toxicity and high efficiency have raised expectations for the development of next-generation protein therapeutics. However, the process of obtaining therapeutic antibodies remains time consuming and empirical. This review summarizes recent progresses in the field of computer-aided antibody development mainly focusing on antibody modeling, which is divided essentially into two parts: (i) modeling the antigen-binding site, also called the complementarity determining regions (CDRs), and (ii) predicting the relative orientations of the variable heavy (VH) and light (VL) chains. Among the six CDR loops, the greatest challenge is predicting the conformation of CDR-H3, which is the most important in antigen recognition. Further computational methods could be used in drug development based on crystal structures or homology models, including antibody–antigen dockings and energy calculations with approximate potential functions. These methods should guide experimental studies to improve the affinities and physicochemical properties of antibodies. Finally, several successful examples of in silico structure-based antibody designs are reviewed. We also briefly review structure-based antigen or immunogen design, with application to rational vaccine development. PMID:22661385

Kuroda, Daisuke; Shirai, Hiroki; Jacobson, Matthew P.; Nakamura, Haruki

2012-01-01

217

FTA-ABS test  

MedlinePLUS

... blood test to detect antibodies to the bacteria Treponema pallidum, which causes syphilis. This test is used when ... Tramont EC. Treponema pallidum (syphilis). In: Mandell GL, Bennett JE, ... and Practice of Infectious Diseases . 7th ed. Philadelphia, ...

218

Antibodies in human infectious disease  

Microsoft Academic Search

Investigation of human antibody responses to viral pathogens at the molecular level is revealing novel aspects of the interplay\\u000a of viruses with the humoral immune system. In viral infection, at least two types of human antibody responses exist: a response\\u000a to mature envelope on virions that is neutralizing and a response to immature forms of envelope (viral debris) that is

Paul W. H. I. Parren; Pascal Poignard; Henrick J. Ditzel; R. Anthony Williamson; Dennis R. Burton

2000-01-01

219

Bispecific Antibodies and Gene Therapy  

Microsoft Academic Search

\\u000a Gene therapy is the transfer of therapeutic genes, via gene transfer vectors, into patients for therapeutic purposes. Different\\u000a gene therapy strategies are being pursued, including long-term gene correction of monogenetic diseases, eradication of tumor\\u000a cells in cancer patients, or genetic vaccination for infectious diseases. Bispecific antibodies and gene therapy are connected\\u000a in two ways. First, bispecific antibodies are tools of

Dirk M. Nettelbeck

220

Human Monoclonal Antibody Against Mesothelin  

Cancer.gov

Mesothelin is a cell surface protein that is naturally expressed at very low levels, but that is significantly increased in aggressive tumors such as mesotheliomas, and pancreatic and ovarian tumors. Therefore, mesothelin is an excellent candidate for tumor-targeted immunotherapeutics. However, the only antibodies against mesothelin that are currently available for clinical trials are of murine origin. The use of these antibodies may be limited by their potential to elicit adverse immune responses in patients with repeated doses.

221

Antibodies to watch in 2014.  

PubMed

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

Reichert, Janice M

2014-01-01

222

Avian Diagnostic and Therapeutic Antibodies  

SciTech Connect

A number of infectious agents have the potential of causing significant clinical symptomology and even death, but dispite this, the number of incidence remain below the level that supports producing a vaccine. Therapeutic antibodies provide a viable treatment option for many of these diseases. We proposed that antibodies derived from West Nile Virus (WNV) immunized geese would be able to treat WNV infection in mammals and potential humans. We demonstrated that WNV specific goose antibodies are indeed successful in treating WNV infection both prophylactically and therapeutically in a golden hamster model. We demonstrated that the goose derived antibodies are non-reactogenic, i.e. do not cause an inflammatory response with multiple exposures in mammals. We also developed both a specific pathogen free facility to house the geese during the antibody production phase and a patent-pending purification process to purify the antibodies to greater than 99% purity. Therefore, the success of these study will allow a cost effective rapidly producible therapeutic toward clinical testing with the necessary infrastructure and processes developed and in place.

Bradley, David Sherman [UND SMHS] [UND SMHS

2012-12-31

223

Selecting and screening recombinant antibody libraries  

Microsoft Academic Search

During the past decade several display methods and other library screening techniques have been developed for isolating monoclonal antibodies (mAbs) from large collections of recombinant antibody fragments. These technologies are now widely exploited to build human antibodies with high affinity and specificity. Clever antibody library designs and selection concepts are now able to identify mAb leads with virtually any specificity.

Hennie R Hoogenboom

2005-01-01

224

Engineered antibodies take center stage.  

PubMed

The start of the post-genomic era provides a useful juncture for reflection on the state of antibody engineering, which will be a critical technology for relating function and pathology to genomic sequence in biology and medicine. The phenomenal progress in deciphering the human genome has given significant impetus to the application of engineered antibodies in proteomics. Thus, advances in phage display antibody libraries can now help to define novel gene function and the measurement of abnormal protein expression in pathological states. Furthermore, intrabody and antibody engineering provide vehicles for the development of molecular medicines of the future. In addition to these new directions, antibody engineering has begun to show concrete success in its long-term efforts to develop targeted immunotherapies for cancer and other diseases. The cornerstones of clinical development are the detailed academic clinical trials that continue to push the boundaries of engineered antibodies into the real world. The field displays a healthy impatience for practical results, as research accelerates with concerted efforts to transfer preclinical insights into clinical trials. Growing private and governmental expenditures will lead to the rapid expansion of life-saving immunotherapeutic agents. The present review developed from our effort to report on the 11th Annual International Conference on Antibody Engineering (3-6 December 2000). This annual meeting is a forum for discussions on the latest advances in antibody engineering groups from around the world, and now includes the broader agenda of engineering in molecular immunology. In bringing scientists together to exchange ideas at this open forum, new collaborations and the threads of new discoveries are woven. For example, Professors Gerhard Wagner (Harvard Medical School), Dennis Burton (Scripps Research Institute), and Peter Hudson (CSIRO, Melbourne, Australia) gave exciting insights on structural immunobiology that had implications across many disciplines. The growth in antibody engineering was highlighted by the attendance of some 600 participants at the meeting, doubling that of the 1999 meeting. Dramatic clinical acceptance of monoclonal antibodies during the past two years has fostered this growth, with sales in 2000 of 1.8 billion dollars and projections for 2001 of 3 billion dollars. However, economic measures cannot begin to convey the medical revolution that is being effected by these first humanized and chimerized monoclonal antibodies. At this juncture, the 10 monoclonal antibody therapeutics in clinical use are of murine origin, of which 3 are entirely murine (OKT3, Mylotarg, 90Y-labeled Bexxar), 4 have been chimerized (human constant domains replacing murine) (ReoPro, Rituxan and its 131I-labeled analogue (Zevalin), Simulect, Remicade) and 3 were chimerized and humanized (human residues being substituted for at least some mouse-specific framework residues in VH and VL) (Zenapax, Herceptin, Synagis). Fully humanized anti-CD52 (CAMPATH-1H) has also been approved by the FDA for the treatment of B-cell chronic lymphocytic leukemia and should become available in late 2001. Humanization was initially developed by Dr. Greg Winter at the MRC Laboratory of Molecular Biology (Cambridge, UK), who presented the meeting's keynote address, "Antibodies as a Paradigm for Molecular Evolution". His pioneering work in antibody phage display libraries has been reformulated into a daring approach to develop truly novel proteins with genetically paired structural elements. He described studies in combinatorial protein engineering with enormous implications for both industrial and therapeutic applications of macromolecules. PMID:11847424

Huston, J S; George, A J

2001-01-01

225

Anti-actin antibodies revealed by counter-immunoelectrophoresis. Relation to smooth muscle antibodies and bile canalicular antibodies.  

PubMed Central

In investigations by counter-immunoelectrophoresis, anti-actin antibodies were found in 59% of patients with chronic hepatitis and in 8% of patients with non-hepatic diseases and normal blood donors. Anti-actin antibodies were found more frequently in patients with hepatitis and IgG smooth muscle antibodies than in other groups of diseases and normal subjects with IgG smooth muscle antibodies. Anti-actin antibodies showed no correlation with bile canalicular antibodies. Images Fig. 1 Fig. 2 PMID:6893598

Diederichsen, H; Riisom, K

1980-01-01

226

Uses of monoclonal antibody 8H9  

DOEpatents

This invention provides a composition comprising an effective amount of monoclonal antibody 8H9 or a derivative thereof and a suitable carrier. This invention provides a pharmaceutical composition comprising an effective amount of monoclonal antibody 8H9 or a derivative thereof and a pharmaceutically acceptable carrier. This invention also provides an antibody other than the monoclonal antibody 8H9 comprising the complementary determining regions of monoclonal antibody 8H9 or a derivative thereof, capable of binding to the same antigen as the monoclonal antibody 8H9. This invention provides a substance capable of competitively inhibiting the binding of monoclonal antibody 8H9. This invention also provides an isolated scFv of monoclonal antibody 8H9 or a derivative thereof. This invention also provides the 8H9 antigen. This invention also provides different uses of the monoclonal antibody 8H9 or its derivative.

Cheung, Nai-Kong V.

2013-04-09

227

Entanglement model of antibody viscosity.  

PubMed

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

Schmit, Jeremy D; He, Feng; Mishra, Shradha; Ketchem, Randal R; Woods, Christopher E; Kerwin, Bruce A

2014-05-15

228

Molecular-specific urokinase antibodies  

NASA Technical Reports Server (NTRS)

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.

Atassi, M. Zouhair (Inventor); Morrison, Dennis R. (Inventor)

2009-01-01

229

Novel Antibody Vectors for Imaging  

PubMed Central

Non-invasive molecular imaging approaches include nuclear, optical, MRI, CT, ultrasound and photoacoustic imaging, which require accumulation of a signal delivered by a probe at the target site. Monoclonal antibodies (mAbs) are high affinity molecules that can be used for specific, high signal delivery to cell surface molecules. However, their long circulation time in blood makes them unsuitable as imaging probes. Efforts to improve antibodies pharmacokinetics without compromising affinity and specificity have been made through protein engineering. Antibody variants that differ in antigen binding sites and size have been generated and evaluated as imaging probes to target tissues of interest. Fast clearing fragments such as single-chain Fv (scFv; 25 kDa) with one antigen binding site (monovalent) demonstrated low accumulation in tumors due the low exposure time to the target. Using scFv as building block to produce larger, bivalent fragments such as scFv dimers (diabodies, 50 kDa) and scFv-fusion proteins (80 kDa minibodies and 105 kDa scFv-Fc) resulted in higher tumor accumulation due to their longer residence time in blood. Imaging studies with these fragments following radiolabeling have demonstrated excellent, high contrast images in gamma cameras and PET scanners. Several studies have also investigated antibody fragments conjugated to fluorescence (near infrared dyes), bioluminescence (luciferases) and quantum dots for optical imaging and iron oxides nanoparticles for MRI. However, these studies indicate that there are several factors that influence successful targeting and imaging. These include stability of the antibody fragment, the labeling chemistry (direct or indirect), whether critical residues are modified, the number of antigen expressed on the cell, and whether the target has a rapid recycling rate or internalizes upon binding. The preclinical data presented are compelling and it is evident that antibody-based molecular imaging tracers will play an important future role in the diagnosis and management of cancer and other diseases. PMID:20350626

Olafsen, Tove; Wu, Anna M.

2010-01-01

230

Trend in prevalence of syphilis among voluntary blood donors in Xi'an, China from 2006 to 2010.  

PubMed

This study was conducted to examine the prevalence of syphilis among blood donors in the Xi'an region of China. All blood donors were unremunerated volunteers recruited from 2006 to 2010. Anti-Treponema pallidum and anti-HIV serology responses were determined using ELISA kits. Among 159 902 voluntary blood donors tested, a total of 575 syphilis (0.36%) and 55 HIV (0.03%) infections were identified. While an increasing trend was shown for the prevalence of both syphilis and HIV over the 5-year period, there was no statistical correlation between the two infections. Our results indicate that syphilis and HIV infections are increasing risk factors for the spread of blood-borne infections. Further investigations and improvements in blood collection and testing procedures are needed to help ensure the safety of donated blood in China. PMID:24291114

Chen, Yaozhen; Liu, Zhixin; Zhang, Qingping; Chen, Jie; Sun, Wengli; Yi, Jing; Zhang, Lingling; Zhao, Peng; Li, Long; Mu, Shijie; Yin, Wen; Zhang, Xianqing; Hu, Xingbin

2014-02-01

231

Rh Antibodies Detectable only by Enzyme Technique  

PubMed Central

The titration of Rh antibodies at intervals during pregnancy by various methods has led to evidence for the existence of an Rh antibody or antibodies detectable by an enzyme (papain) method but not by anti-human-globulin. Adsorption experiments show that this antibody is less readily absorbed by red cells than the other Rh antibodies unless the cells are pretreated with enzyme. This fact may possibly account for the finding of a negative or weakly positive anti-human-globulin test associated with moderate or high titres by enzyme techniques. The relationship of this antibody to the general immunization process in pregnant women is discussed. PMID:13886834

Dodd, Barbara E.; Eeles, Doreen A.

1961-01-01

232

Emerging antibody combinations in oncology  

PubMed Central

The use of monoclonal antibodies (mAbs) has become a general approach for specifically targeting and treating human disease. In oncology, the therapeutic utility of mAbs is usually evaluated in the context of treatment with standard of care, as well as other small molecule targeted therapies. Many anti-cancer antibody modalities have achieved validation, including the targeting of growth factor and angiogenesis pathways, the induction of tumor cell killing or apoptosis and the blocking of immune inhibitory mechanisms to stimulate anti-tumor responses. But, as with other targeted therapies, few antibodies are curative because of biological complexities that underlie tumor formation and redundancies in molecular pathways that enable tumors to adapt and show resistance to treatment. This review discusses the combinations of antibody therapeutics that are emerging to improve efficacy and durability within a specific biological mechanism (e.g., immunomodulation or the inhibition of angiogenesis) and across multiple biological pathways (e.g., inhibition of tumor growth and induction of tumor cell apoptosis). PMID:21697653

Hariharan, Kandasamy

2011-01-01

233

Monoclonal antibodies: application in radiopharmacy.  

PubMed

In this study was carried on a systematic review of the data was carried out in the topic of monoclonal antibodies in the last 40 years. All the data collected and summarized revealed that this new class of medicine may bring great advance in the field of radiopharmacy, oncology and imaging. PMID:24251731

Ligiero, Thais Braga; de Souza Albernaz, Marta; de Carvalho, Samira Marques; de Oliveira, Silvia Maria Velasques; Santos-Oliveira, Ralph

2013-12-01

234

Rabbit polyclonal antibody to Peripherin  

Microsoft Academic Search

Peripherin is a Class III intermediate filament subunit found in both the peripheral and central nervous systems, though it is concentrated, as its name suggests, in the neurons of peripheral ganglia and their processes. Antibodies to peripherin can be used in identifying, classifying, and studying neurons in the nervous system. Peripherin is also a good diagnostic marker for ballooned axons

Species Reactivity

2006-01-01

235

Novel antibodies as anticancer agents  

Microsoft Academic Search

In recent years antibodies, whether generated by traditional hybridoma technology or by recombinant DNA strategies, have evolved from Paul Ehrlich's ‘magic bullets’ to a modern age ‘guided missile’. In the recent years of immunologic research, we are witnessing development in the fields of antigen screening and protein engineering in order to create specific anticancer remedies. The developments in the field

I Zafir-Lavie; Y Michaeli; Y Reiter

2007-01-01

236

Detection of Campylobacter species using monoclonal antibodies  

NASA Astrophysics Data System (ADS)

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.

Young, Colin R.; Lee, Alice; Stanker, Larry H.

1999-01-01

237

2006 Nature Publishing Group Department of Antibody  

E-print Network

and the second largest class of drugs after vaccines. The generation of potent antibody therapeutics, which I, which further increases the cost. Although antibody therapeutics are often safe and well tolerated, rare

Cai, Long

238

Towards a carbon nanotube antibody sensor  

E-print Network

This work investigated single-walled carbon nanotube (SWNT)/polymer-protein A complexes for optically reporting antibody concentration via a change in near infrared fluorescent emission after antibody binding. SWNT have ...

Bojö, Peter

2012-01-01

239

A Monoclonal Antibody Against PMEL.  

PubMed

PMEL, also known as Pmel17 or gp100, is a melanocyte-specific glycoprotein that is essential for the formation of stage II melanosomes. As it has a highly restricted expression pattern in normal tissues and a transient presence on the cell surface, PMEL is believed to be a potential target for antibody drug conjugate therapy in some pigmentary diseases. The production of a high specificity and high affinity monoclonal antibody against human PMEL was helpful for the antibody drug conjugate therapy study. In the present study, monoclonal antibodies (MAbs) against PMEL were obtained by immunizing BALB/c mice with the recombinant PMEL-GST fusion protein. Three mAbs (A3F, G11B, and J7E) with a titer of 1:6000, 1:10,000, and 1:3000, respectively, were obtained. Immunoglobulin subclass assay revealed that A3F was IgG2b, G11B was IgG1, and J7E was IgG2a. Specificity analysis by Western blotting demonstrated that A3F and J7E cross-reacted with GPNMB or LAMP; however, G11B reacted with PMEL only. Immunohistochemistry experiments showed that G11B could bind human PMEL antigen in normal skin. Flow cytometry assay demonstrated that G11B could bind to the surface of PMEL positive melanoma cells but not PMEL negative cells. Taken together, these results show that this G11B provides a useful tool for the antibody drug conjugate therapy study in some pigmentary diseases. PMID:25118787

Shi, Fangyuan; Xu, Zhenjie; Chen, Hongdong; Wang, Xin; Cui, Jihong; Zhang, Ping; Zhang, Ping; Xie, Xin

2014-10-01

240

Antibody response to Rocky Mountain spotted fever.  

PubMed Central

Various techniques were compared to determine the most sensitive method for detection of rocky Mountain spotted fever antibody. A radiometabolic technique for detection of Rocky Mountain spotted fever antibody is also described. In infected monkeys, the fluorescent antibody technique yielded the earliest evidence of seroconversion; with some monkeys the microagglutination procedure was equally effective. The fluorescent antibody and microagglutination measurements showed higher titers than those for complement fixation, Weil-Felix, or the radiometabolic techniques. PMID:819455

Kenyon, R H; Canonico, P G; Sammons, L S; Bagley, L R; Pedersen, C E

1976-01-01

241

Original article Monoclonal antibodies against goldfish  

E-print Network

immunoglobulin and antibody levels by ELISA in carp (Cyprinus carpio) AK Siwicki C Vergnet J Charlemagne M Dunier decreased until day 28. monoclonal antibody / ELISA / ELISPOT / antibody-secreting cells / Cyprinus carpio suite. anticorps monoc/ona//EL/S/Cyprinus carpio

Paris-Sud XI, Université de

242

Induction and detection of antibodies to squalene  

Microsoft Academic Search

An enzyme-linked immunosorbent assay (ELISA) utilizing antigen coated on hydrophobic polyvinyldiene fluoride (PVDF) membranes is described for detecting antibodies that bind to squalene (SQE). Because of the prior lack of availability of validated antibodies to SQE, positive controls for the assay were made by immunization with formulations containing SQE to create monoclonal antibodies (mAbs) that reacted with SQE. Among eight

Gary R Matyas; Nabila M Wassef; Mangala Rao; Carl R Alving

2000-01-01

243

Anti-DNA antibodies in SLE  

SciTech Connect

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.

Voss, E.W.

1988-01-01

244

Antibody immobilization using heterobifunctional crosslinkers  

Microsoft Academic Search

Covalent attachment of functional proteins to a solid support is important for biosensors. One method employs thiol-terminal silanes and heterobifunctional crosslinkers such as N-succinimidyl 4-maleimidobutyrate (GMBS) to immobilize proteins through amino groups onto glass, silica, silicon or platinum surfaces. In this report, several heterobifunctional crosslinkers are compared to GMBS for their ability to immobilize active antibodies onto glass cover slips

Lisa C. Shriver-Lake; Brian Donner; Rebecca Edelstein; Kristen Breslin; Suresh K. Bhatia; Frances S. Ligler

1997-01-01

245

Single-Chain Antibody Library  

DOE Data Explorer

Researchers at Pacific Northwest National Laboratory (PNNL) have constructed a nonimmune library consisting of 109 human antibody scFv fragments, which have been cloned and expressed on the surface of yeast. Nanomolar-affinity scFvs are routinely obtained by magnetic bead screening and flow cytometric sorting. The yeast library can be amplified 1010 fold without measurable loss of clonal diversity. This allows for indefinite expansion of the library. All scFv clones can be assessed directly on the yeast cell surface by immunofluorescent labeling and flow cytometry, obviating separate subcloning, expression, and purification steps. The ability to use multiplex library screening demonstrates the utility of this approach for high-throughput antibody isolation for proteomic applications. The yeast library may be used for research projects or teaching performed for U.S. Government purposes only. If you would like to request an aliquot of the single-chain antibody library for your research, please print and fill out the Materials Transfer Agreement (MTA) [PDF, 20K]. The website provides the contact information for mailing the MTA. [copied from http://www.sysbio.org/dataresources/singlechain.stm

Baird, Cheryl

246

Improved monoclonal antibodies to halodeoxyuridine  

DOEpatents

The development, method of production, characterization and methods of use of two hybridomas, CIdU-1 (ATCC Accession No. HB-8321) and CIdU-2 (ATCC Accession No. HB-8320), are described. These secrete IgG/sub 1/(K) immunoglobulins that react with halodeoxyuridine (HdU or halodU) such as bromo, chloro, fluoro and iodo deoxyuridine (BrdU, CldU, FdU and IdU), whether these are free in solution or incorporated into single stranded DNA in whole cells. The antibodies do not react with naturally occurring free nucleic acids or with deoxyribonucleic acid (DNA) or ribonucleic acid (RNA) polymers. These antibodies are suitable for use in enzyme immunoassays for free CldU, FdU, IdU and BrdU and for detecting cells with these nucleotides incorporated into them. The monoclonal antibodies are useful in the detection of the sensitivity of tumor cells to specific chemotherapeutic agents, in the measurement of the rate of cellular DNA synthesis, in the measurement of the rate of proliferation of normal and malignant cells and in the detection of HPRT deficiency in cells. 1 tab.

Vanderlaan, M.; Dolbeare, F.A.; Gray, J.W.; Thomas, C.B.

1983-10-18

247

A Dual Point-of-Care Test Shows Good Performance in Simultaneously Detecting Nontreponemal and Treponemal Antibodies in Patients With Syphilis: A Multisite Evaluation Study in China  

PubMed Central

Background.?Rapid point-of-care (POC) syphilis tests based on simultaneous detection of treponemal and nontreponemal antibodies (dual POC tests) offer the opportunity to increase coverage of syphilis screening and treatment. This study aimed to conduct a multisite performance evaluation of a dual POC syphilis test in China. Methods.?Participants were recruited from patients at sexually transmitted infection clinics and high-risk groups in outreach settings in 6 sites in China. Three kinds of specimens (whole blood [WB], fingerprick blood [FB], and blood plasma [BP]) were used for evaluating sensitivity and specificity of the Dual Path Platform (DPP) Syphilis Screen and Confirm test using its treponemal and nontreponemal lines to compare Treponema pallidum particle agglutination (TPPA) assay and toluidine red unheated serum test (TRUST) as reference standards. Results.?A total of 3134 specimens (WB 1323, FB 488, and BP 1323) from 1323 individuals were collected. The sensitivities as compared with TPPA were 96.7% for WB, 96.4% for FB, and 94.6% for BP, and the specificities were 99.3%, 99.1%, and 99.6%, respectively. The sensitivities as compared with TRUST were 87.2% for WB, 85.8% for FB, and 88.4% for BP, and the specificities were 94.4%, 96.1%, and 95.0%, respectively. For specimens with a TRUST titer of 1:4 or higher, the sensitivities were 100.0% for WB, 97.8% for FB, and 99.6% for BP. Conclusions.?DPP test shows good sensitivity and specificity in detecting treponemal and nontreponemal antibodies in 3 kinds of specimens. It is hoped that this assay can be considered as an alternative in the diagnosis of syphilis, particularly in resource-limited areas. PMID:23132172

Yin, Yue-Ping; Chen, Xiang-Sheng; Wei, Wan-Hui; Gong, Kuang-Long; Cao, Wen-Ling; Yong, Gang; Feng, Liang; Huang, Shu-Jie; Wang, Dong-Mei; Han, Yan; Chen, Shao-Chun; Mabey, David; Peeling, Rosanna W.

2013-01-01

248

IBC's 23rd Antibody Engineering and 10th Antibody Therapeutics Conferences and the Annual Meeting of The Antibody Society  

PubMed Central

Now in its 23rd and 10th years, respectively, the Antibody Engineering and Antibody Therapeutics conferences are the Annual Meeting of The Antibody Society. The scientific program covers the full spectrum of challenges in antibody research and development from basic science through clinical development. In this preview of the conferences, the chairs provide their thoughts on sessions that will allow participants to track emerging trends in (1) the development of next-generation immunomodulatory antibodies; (2) the complexity of the environment in which antibodies must function; (3) antibody-targeted central nervous system (CNS) therapies that cross the blood brain barrier; (4) the extension of antibody half-life for improved efficacy and pharmacokinetics (PK)/pharmacodynamics (PD); and (5) the application of next generation DNA sequencing to accelerate antibody research. A pre-conference workshop on Sunday, December 2, 2012 will update participants on recent intellectual property (IP) law changes that affect antibody research, including biosimilar legislation, the America Invents Act and recent court cases. Keynote presentations will be given by Andreas Plückthun (University of Zürich), who will speak on engineering receptor ligands with powerful cellular responses; Gregory Friberg (Amgen Inc.), who will provide clinical updates of bispecific antibodies; James D. Marks (University of California, San Francisco), who will discuss a systems approach to generating tumor targeting antibodies; Dario Neri (Swiss Federal Institute of Technology Zürich), who will speak about delivering immune modulators at the sites of disease; William M. Pardridge (University of California, Los Angeles), who will discuss delivery across the blood-brain barrier; and Peter Senter (Seattle Genetics, Inc.), who will present his vision for the future of antibody-drug conjugates. For more information on these meetings or to register to attend, please visit www.IBCLifeSciences.com/AntibodyEng or call 800-390-4078. Members of The Antibody Society and mAbs journal subscribers receive a 20% discount for meeting registration. To obtain this discount, email kdostie@ibcusa.com. mAbs is the official therapeutics journal of The Antibody Society and offers a discounted subscription to Society members for $49. PMID:23007482

Marquardt, John; Begent, Richard H.J.; Chester, Kerry; Huston, James S.; Bradbury, Andrew; Scott, Jamie K.; Thorpe, Philip E.; Veldman, Trudi; Reichert, Janice M.; Weiner, Louis M.

2012-01-01

249

Analysis of anticholesterol antibodies using hydrophobic membranes  

Microsoft Academic Search

An analytical immunoblotting procedure and a serological enzyme-linked immunosorbent assay (ELISA) for the characterization of antibodies to cholesterol are described. Hydrophobic membranes consisting of polyvinylidene fluoride (PVDF) are used to immobilize cholesterol for immunodetection by anti-sterol antibodies. To determine whether antibodies to cholesterol were induced after immunization with liposomal cholesterol, we separated total lipid extracts of very-low density lipoproteins by

Jacinta Aniagolu; Glenn M. Swartz; Jan Dijkstra; John W. Madsen; Jennifer J. Raney; Shawn J. Green

1995-01-01

250

Reactivity of Langerhans cells with hybridoma antibody.  

PubMed Central

Reactivity of a monoclonal antibody with human Langerhans cells was demonstrated by a double-labeling immunofluorescence technique. Ia-bearing cells of the epidermis (Langerhans cells) were reactive with this antibody both in frozen sections and in cell suspensions prepared from human epidermis. This monoclonal antibody was unreactive with non-Ia-bearing epidermal cells and with peripheral blood B cells, T cells, and monocytes but did not bind to 70% of intrathymic lymphocytes. These observations further distinguish Langerhans cells from classical monocytes. Furthermore, this monoclonal antibody is a highly specific marker for the in vivo identification and in vitro isolation of Langerhans cells. Images PMID:6941307

Fithian, E; Kung, P; Goldstein, G; Rubenfeld, M; Fenoglio, C; Edelson, R

1981-01-01

251

Reconciling the structural attributes of avian antibodies.  

PubMed

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

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

252

6th Annual European Antibody Congress 2010  

PubMed Central

The 6th European Antibody Congress (EAC), organized by Terrapinn Ltd., was held in Geneva, Switzerland, which was also the location of the 4th and 5th EAC.1,2 As was the case in 2008 and 2009, the EAC was again the largest antibody congress held in Europe, drawing nearly 250 delegates in 2010. Numerous pharmaceutical and biopharmaceutical companies active in the field of therapeutic antibody development were represented, as were start-up and academic organizations and representatives from the US Food and Drug Administration (FDA). The global trends in antibody research and development were discussed, including success stories of recent marketing authorizations of golimumab (Simponi®) and canakinumab (Ilaris®) by Johnson & Johnson and Novartis, respectively, updates on antibodies in late clinical development (obinutuzumab/GA101, farletuzumab/MORAb-003 and itolizumab/T1 h, by Glycart/Roche, Morphotek and Biocon, respectively) and success rates for this fast-expanding class of therapeutics (Tufts Center for the Study of Drug Development). Case studies covering clinical progress of girentuximab (Wilex), evaluation of panobacumab (Kenta Biotech), characterization of therapeutic antibody candidates by protein microarrays (Protagen), antibody-drug conjugates (sanofi-aventis, ImmunoGen, Seattle Genetics, Wyeth/Pfizer), radio-immunoconjugates (Bayer Schering Pharma, Université de Nantes) and new scaffolds (Ablynx, AdAlta, Domantis/GlaxoSmithKline, Fresenius, Molecular Partners, Pieris, Scil Proteins, Pfizer, University of Zurich) were presented. Major antibody structural improvements were showcased, including the latest selection engineering of the best isotypes (Abbott, Pfizer, Pierre Fabre), hinge domain (Pierre Fabre), dual antibodies (Abbott), IgG-like bispecific antibodies (Biogen Idec), antibody epitope mapping case studies (Eli Lilly), insights in Fc?RII receptor (University of Cambridge), as well as novel tools for antibody fragmentation (Genovis). Improvements of antibody druggability (Abbott, Bayer, Pierre Fabre, Merrimack, Pfizer), enhancing IgG pharmacokinetics (Abbott, Chugai), progress in manufacturing (Genmab, Icosagen Cell Factory, Lonza, Pierre Fabre) and the development of biosimilar antibodies (Biocon, Sandoz, Triskel) were also discussed. Last but not least, identification of monoclonal antibodies (mAbs) against new therapeutic targets (Genentech, Genmab, Imclone/Lilly, Vaccinex) including Notch, cMet, TGF?RII, SEMA4D, novel development in immunotherapy and prophylaxis against influenza (Crucell), anti-tumor activity of immunostimulatory antibodies (MedImmune/Astra Zeneca) and translations to clinical studies including immunogenicity issues (Amgen, Novartis, University of Debrecen) were presented. PMID:21441785

2011-01-01

253

Uses of monoclonal antibody 8H9  

DOEpatents

This invention provides a composition comprising an effective amount of monoclonal antibody 8H9 or a derivative thereof and a suitable carrier. This invention provides a pharmaceutical composition comprising an effective amount of monoclonal antibody 8H9 or a derivative thereof and a pharmaceutically acceptable carrier. This invention also provides an antibody other than the monoclonal antibody 8H9 comprising the complementary determining regions of monoclonal antibody 8H9 or a derivative thereof, capable of binding to the same antigen as the monoclonal antibody 8H9. This invention provides a substance capable of competitively inhibiting the binding of monoclonal antibody 8H9. This invention also provides an isolated scFv of monoclonal antibody 8H9 or a derivative thereof. This invention also provides the 8H9 antigen. This invention also provides a method of inhibiting the growth of tumor cells comprising contacting said tumor cells with an appropriate amount of monoclonal antibody 8H9 or a derivative thereof.

Cheung, Nai-Kong V

2013-08-06

254

[Managing patients with therapeutic antibodies in odontostomatology].  

PubMed

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

Demoersman, J; Soueidan, A; Corre, P; Pers, J O

2014-06-01

255

The production of antibody fragments and antibody fusion proteins by yeasts and filamentous fungi  

Microsoft Academic Search

In this review we will focus on the current status and views concerning the production of antibody fragments and antibody fusion proteins by yeasts and filamentous fungi. We will focus on single-chain antibody fragment production (scFv and VHH) by these lower eukaryotes and the possible applications of these proteins. Also the coupling of fragments to relevant enzymes or other components

Vivi Joosten; Christien Lokman; Cees AMJJ van den Hondel; Peter J Punt

2003-01-01

256

A Dynamic Landscape for Antibody Binding Modulates Antibody-Mediated Neutralization of West Nile Virus  

Microsoft Academic Search

Neutralizing antibodies are a significant component of the host's protective response against flavivirus infection. Neutralization of flaviviruses occurs when individual virions are engaged by antibodies with a stoichiometry that exceeds a required threshold. From this “multiple-hit” perspective, the neutralizing activity of antibodies is governed by the affinity with which it binds its epitope and the number of times this determinant

Kimberly A. Dowd; Christiane A. Jost; Anna P. Durbin; Stephen S. Whitehead; Theodore C. Pierson

2011-01-01

257

Asthma and antibodies to pneumococcal virulence proteins  

PubMed Central

Purpose We previously reported that asthmatics had lower anti-serotype-specific pneumococcal polysaccharide antibody levels than non-asthmatics, and T-helper 2 (Th2)-immune profile was associated with suboptimal pneumococcal polysaccharide antibody. Our objective was to determine the influence of asthma status on anti-pneumococcal protein antigen antibody levels. Methods We conducted a cross-sectional study, which enrolled 16 children and adults with asthma and 14 subjects without asthma. Asthma was ascertained by predetermined criteria. Serum IgG antibody levels to pneumococcal surface protein A (PspA), pneumococcal surface protein C (PspC), pneumococcal choline-binding protein A (PcpA), and pneumolysin (PLY) were measured by ELISA assays. These antibody levels were compared between asthmatics and non-asthmatics. Th2-immune profile was determined by IL-5 secretion from PBMCs cultured with house dust mite (HDM) and staphylococcal enterotoxin B (SEB) at day seven. The correlation between the anti-pneumococcal antibody levels and Th2-HDM and SEB-responsive immune profile was assessed. Results Of the 30 subjects, 16 (53%) were male and the median age was 26 years. There were no significant differences in anti-PspA, anti-PspC, anti-PcpA, and anti-PLY antibody levels between asthmatics and non-asthmatics. Th2-immune profile was inversely correlated with anti-PspC antibody levels (r= ?0.53, p=0.003). This correlation was significantly modified by asthma status (r= ?0.74, p=0.001 for asthmatics vs. r= ?0.06, p=0.83 for non-asthmatics). Other pneumococcal protein antibodies were not correlated with Th2-immune profile. Conclusion No significant differences in anti-pneumococcal protein antigen antibody levels between asthmatics and non-asthmatics were found. Asthma status is an important effect modifier determining the negative influence of Th2-immune profile on anti-PspC antibody levels. PMID:23749296

Zhao, Hongxia; Jung, Ji A; Briles, David E.; Kita, Hirohito; Tsigrelis, Constantine; Juhn, Young J.

2013-01-01

258

Engineered antibody fragments and the rise of single domains  

Microsoft Academic Search

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))

Philipp Holliger; Peter J Hudson

2005-01-01

259

Transplacental Chikungunya Virus Antibody Kinetics, Thailand  

PubMed Central

Antibodies to chikungunya virus were detected by hemagglutination-inhibition assay in 33.6% of 2,000 infants' cord sera at delivery. Follow-up of 24 seropositive infants showed that the half-life of antibody persistence was 35.5 days. Chikungunya virus infection is common in Thailand, and routine use of diagnostic assays is needed. PMID:17283634

Endy, Timothy P.; Simasathien, Sriluck; Kerdpanich, Angkool; Polprasert, Napuschon; Aree, Chanchai; Vaughn, David W.; Nisalak, Ananda

2006-01-01

260

The therapeutic antibodies market to 2008.  

PubMed

The therapeutic biologics market is currently dominated by recombinant protein products. However, many of these products are mature, and growth of the biologics market will increasingly rely on the expansion of the therapeutic monoclonal antibody sector. Successive technology waves have driven the growth of the monoclonal antibody sector, which is currently dominated by chimeric antibodies. Chimeric products, led by Remicade and Rituxan, will continue to drive market share through to 2008. However, over the forecast period, humanized and fully human monoclonal antibodies, together with technologies such as Fabs and conjugated antibodies, will play an increasingly important role, driving monoclonal antibody market growth at a forecast compound annual growth rate of 20.9%, to reach $16.7 billion by 2008. In terms of therapeutic focus, the monoclonal antibody market is heavily focused on oncology and arthritis, immune and inflammatory disorders, and products within these therapeutic areas are set to continue to be the key growth drivers over the forecast period. Underlying the growth of the market is the evolution of the monoclonal antibody company business model, set to transition towards the highly successful innovator model. PMID:15760719

Pavlou, Alex K; Belsey, Mark J

2005-04-01

261

Cytolytic Antibodies to Melanocytes in Vitiligo  

Microsoft Academic Search

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.

Jian Cui; Yuko Arita; Jean-Claude Bystryn

1993-01-01

262

7th Annual European Antibody Congress 2011  

PubMed Central

The 7th European Antibody Congress (EAC), organized by Terrapin Ltd., was again held in Geneva, Switzerland, following on the tradition established with the 4th EAC. The 2011 version of the EAC was attended by nearly 250 delegates who learned of the latest advances and trends in the global development of antibody-based therapeutics. The first day focused on advances in understanding structure-function relationships, choosing the best format, glycoengineering biobetter antibodies, improving the efficacy and drugability of mAbs and epitope mapping. On the second day, the discovery of novel targets for mAb therapy, clinical pipeline updates, use of antibody combinations to address resistance, generation and identification of mAbs against new targets and biosimilar mAb development were discussed. Antibody-drug conjugates, domain antibodies and new scaffolds and bispecific antibodies were the topics of the third day. In total, nearly 50 speakers provided updates of programs related to antibody research and development on-going in the academic, government and commercial sectors. PMID:22453093

2012-01-01

263

Antibody neutralization and escape by HIV1  

Microsoft Academic Search

Neutralizing antibodies (Nab) are a principal component of an effective human immune response to many pathogens, yet their role in HIV-1 infection is unclear. To gain a better understanding of this role, we examined plasma from patients with acute HIV infection. Here we report the detection of autologous Nab as early as 52 days after detection of HIV-specific antibodies. The

Xiping Wei; Julie M. Decker; Shuyi Wang; Huxiong Hui; John C. Kappes; Xiaoyun Wu; Jesus F. Salazar-Gonzalez; Maria G. Salazar; J. Michael Kilby; Michael S. Saag; Natalia L. Komarova; Martin A. Nowak; Beatrice H. Hahn; Peter D. Kwong; George M. Shaw

2003-01-01

264

Antireticulin antibody: Incidence and diagnostic significance  

Microsoft Academic Search

Sera from 101 patients with adult coeliac disease, 46 patients with childhood coeliac disease, 50 patients with dermatitis herpetiformis, and 479 patients with various other diseases, including skin, gastrointestinal, haematological, and immunological disorders, have been tested for the presence of the antireticulin antibody. Positive sera were retested at higher dilutions. Antireticulin antibody was only found in a significant proportion of

P. P. Seah; Lionel Fry; E. J. Holborow; Mary A. Rossiter; W. F. Doe; A. F. Magalhaes; A. V. Hoffbrand

1973-01-01

265

Production of human monoclonal antibodies to myeloperoxidase.  

PubMed Central

Two mouse-human heterohybridomas secreting human antibodies to myeloperoxidase (MPO) were derived from the peripheral blood of a patient who developed microscopic polyarteritis as the result of long-term treatment with hydralazine. Forty-five immunoglobulin-secreting lines were obtained from the fusion of patient lymphocytes with the CB-F7 heteromyeloma cell line. Of these, two antibodies, one IgG and one IgM, bound to myeloperoxidase in solid phase ELISA and gave a perinuclear staining pattern on ethanol-fixed human neutrophil cytospin preparations. The staining patterns were similar to those seen with serum from the patient. Antigen-inhibition studies revealed that the affinity of the IgG monoclonal antibody was 28 times higher (k = 1.4 x 10(-7)) than the IgM antibody (k = 5 x 10(-5)). Cross-inhibition studies further suggested that the two monoclonal antibodies recognized the same epitope on MPO. Of the other secreting cell lines, none produced antibody which reacted with the panel of autoantigens used for testing. Neither mononuclear antibody reacted with this panel indicating that they were not simply polyreactive natural autoantibodies. These are the first human monoclonal antibodies to native myeloperoxidase to be reported. Images Figure 2 Figure 3 PMID:1337335

Ehrenstein, M R; Leaker, B; Isenberg, D; Cambridge, G

1992-01-01

266

Thyroglobulin Antibodies in Differentiated Thyroid Cancer  

Microsoft Academic Search

A retrospective review of patients with differentiated thyroid cancer (DTC) who were seen between 1984 and 1996 at the Royal Marsden Hospital identified 40 patients with serum thyroglobulin antibodies (TgAb). These antibodies can interfere with the immunoradiometric assay for serum thyroglobulin (Tg) used at this hospital, with resulting underestimation of the Tg level. A review of the case notes was

P. Hjiyiannakis; J. Mundy; C. Harmer

1999-01-01

267

Fixation of Complement to Fragments of Antibody  

Microsoft Academic Search

A specific precipitate of ovalbumin and its rabbit-serum antibody, after fixing human serum complement, was digested with papain. The digest was analyzed immunochemically for complexes of antigen, antibody fragments, and components of complement. The results indicated that complement is not bound to Porter fragment III, but very likely is bound to fragments I and II.

A. M. Reiss; O. J. Plescia

1963-01-01

268

8th Annual European Antibody Congress 2012  

PubMed Central

The 8th European Antibody Congress (EAC), organized by Terrapin Ltd., was again held in Geneva, Switzerland, following on the tradition established with the 4th EAC. The new agenda format for 2012 included three parallel tracks on: (1) naked antibodies; (2) antibody drug conjugates (ADCs); and (3) bispecific antibodies and alternative scaffolds. The meeting started and closed with three plenary lectures to give common background and to share the final panel discussion and conclusions. The two day event included case studies and networking for nearly 250 delegates who learned of the latest advances and trends in the global development of antibody-based therapeutics. The monoclonal antibody track was focused on understanding the structure-function relationships, optimization of antibody design and developability, and processes that allow better therapeutic candidates to move through the clinic. Discussions on novel target identification and validation were also included. The ADC track was dedicated to evaluation of the ongoing success of the established ADC formats alongside the rise of the next generation drug-conjugates. The bispecific and alternative scaffold track was focused on taking stock of the multitude of bispecific formats being investigated and gaining insight into recent innovations and advancements. Mechanistic understanding, progression into the clinic and the exploration of multispecifics, redirected T cell killing and alternative scaffolds were extensively discussed. In total, nearly 50 speakers provided updates of programs related to antibody research and development on-going in the academic, government and commercial sectors. PMID:23493119

Beck, Alain; Carter, Paul J.; Gerber, Hans-Peter; Lugovskoy, Alexey A.; Wurch, Thierry; Junutula, Jagath R.; Kontermann, Roland E; Mabry, Robert

2013-01-01

269

Monoclonal antibodies to rat renal antigens.  

PubMed Central

We have developed hybridoma cell lines which secrete monoclonal antibodies to some rat renal antigens, namely the brush border of proximal tubular epithelium and the cytoplasm of tubular cells. The immunoglobulin class of the hybridoma was found to be IgG1. Specific antibody activity against either glomerular basement membrane (GBM) and tubular basement membrane (TBM) or Bowman's capsule and a part of TBM was observed, although these hybridoma cell lines have not yet been successfully established. In particular, the hybridoma secreting antibodies to TBM did not remain stable during antibody production, and was lost during the culture and cloning procedures. These monoclonal antibodies should be of value in research on the pathogenesis of human glomerulonephritis. Images Figure 1 Figure 2 PMID:6376337

Shimizu, F; Orikasa, M; Sato, K; Oite, T

1984-01-01

270

Dual targeting strategies with bispecific antibodies  

PubMed Central

Monoclonal antibodies are widely used for the treatment of cancer, inflammatory and infectious diseases and other disorders. Most of the marketed antibodies are monospecific and therefore capable of interacting and interfering with a single target. However, complex diseases are often multifactorial in nature, and involve redundant or synergistic action of disease mediators or upregulation of different receptors, including crosstalk between their signaling networks. Consequently, blockade of multiple, different pathological factors and pathways may result in improved therapeutic efficacy. This result can be achieved by combining different drugs, or use of the dual targeting strategies applying bispecific antibodies that have emerged as an alternative to combination therapy. This review discusses the various dual targeting strategies for which bispecific antibodies have been developed and provides an overview of the established bispecific antibody formats. PMID:22453100

2012-01-01

271

Viral antibodies in coyotes from California.  

PubMed

Prevalence of antibodies against canine parvovirus (CPV), canine distemper virus (CDV), and canine adenovirus type 1 (CAV) were determined among 152 coyotes (Canis latrans) at the Naval Petroleum Reserves (NPRC; California, USA) from 1985 to 1990. Overall prevalence of antibodies to CPV, CDV, and CAV was 66%, 37%, and 68%, respectively. Prevalence of CPV and CDV varied significantly among years. Antibody prevalence did not differ between sexes for any disease, but did vary significantly among age classes and was lowest for pups (< 1-yr-old). Among pups, antibody prevalence increased with age for all three diseases. Coyotes are a potential source of viral exposure for endangered San Joaquin kit foxes (Vulpes macrotis mutica), but variation in coyote abundance did not appear to influence antibody prevalence among kit foxes. PMID:9577772

Cypher, B L; Scrivner, J H; Hammer, K L; O'Farrell, T P

1998-04-01

272

Antibody-Mediated Lung Transplant Rejection  

PubMed Central

Antibody-mediated rejection after lung transplantation remains enigmatic. However, emerging evidence over the past several years suggests that humoral immunity plays an important role in allograft rejection. Indeed, the development of donor-specific antibodies after transplantation has been identified as an independent risk factor for acute cellular rejection and bronchiolitis obliterans syndrome. Furthermore, cases of acute antibody-mediated rejection resulting in severe allograft dysfunction have been reported, and these demonstrate that antibodies can directly injure the allograft. However, the incidence and toll of antibody-mediated rejection are unknown because there is no widely accepted definition and some cases may be unrecognized. Clearly, humoral immunity has become an important area for research and clinical investigation. PMID:23002428

Hachem, Ramsey

2012-01-01

273

Antibody to intermediate filaments of the cytoskeleton.  

PubMed Central

IgM antibodies against cultures of intermediate filaments (IMF) of the cytoskeleton were demonstrated by immunofluorescence in the sera of 94 (80%) of 118 patients with seropositive rheumatoid arthritis. These antibodies reacted with IMF in cultures of both human fetal fibroblasts and laryngeal carcinoma (HEp2) cells. Of 10 patients from whom paired synovial fluids were also available 8 had anti-IMF antibodies in both serum and fluid. In seronegative RA the incidence of anti-IMF was 40%, in ankylosing spondylitis 25%, in osteoarthrosis 16%, and in normal subjects 14%. Only a minority of RA sera positive for anti-IMF antibodies were also positive for smooth muscle antibody. Absorption experiments suggest that in RA anti-IMF is directed at the intermediate filament protein, vimentin. Images PMID:7039524

Osung, O A; Chandra, M; Holborow, E J

1982-01-01

274

PROPERTIES OF ANTIBODIES CYTOPHILIC FOR MACROPHAGES  

PubMed Central

Cytophilic activity for macrophages was shown to be a property possessed by most, if not all, of the complement binding 7S ?2-population of guinea pig antibodies. Cytophilic antibodies were also demonstrated in rabbit and mouse antisera to sheep red cells. While each species antibody best sensitized homologous macrophages, cross species sensitization was also observed. The binding site for macrophages resides on the Fc fragment, therefore, on the H chains, and is destroyed by pepsin hydrolysis or reduction and alkylation. The binding reaction is reversible with a high rate of dissociation at 37°C. Cytophilic activity is not complement dependent, and was shown to be that property of opsonizing antibody which provides the receptors that permit the binding of the antibody to the macrophage cell membrane in preparation for phagocytosis. PMID:4159249

Berken, Arthur; Benacerraf, Baruj

1966-01-01

275

Rapid selection of cell subpopulation-specific human monoclonal antibodies from a synthetic phage antibody library.  

PubMed Central

Peripheral blood leukocytes incubated with a semisynthetic phage antibody library and fluorochrome-labeled CD3 and CD20 antibodies were used to isolate human single-chain Fv antibodies specific for subsets of blood leukocytes by flow cytometry. Isolated phage antibodies showed exclusive binding to the subpopulation used for selection or displayed additional binding to a restricted population of other cells in the mixture. At least two phage antibodies appeared to display hitherto-unknown staining patterns of B-lineage cells. This approach provides a subtractive procedure to rapidly obtain human antibodies against known and novel surface antigens in their native configuration, expressed on phenotypically defined subpopulations of cells. This approach does not depend on immunization procedures or the necessity to repeatedly construct phage antibody libraries. Images Fig. 1 PMID:7537380

de Kruif, J; Terstappen, L; Boel, E; Logtenberg, T

1995-01-01

276

Immobilization of antibodies and enzyme-labeled antibodies by radiation polymerization  

SciTech Connect

Immobilization of antibodies and enzyme-labeled antibodies by radiation polymerization at low temperatures was studied. The antibody activity of antibody was not affected by irradiation at an irradiation dose of below 8 MR and low temperatures. Immobilization of peroxidase-labeled anti-rabbit IgG goat IgG, anti-peroxidase, peroxidase, and anti-alpha-fetoprotein was carried out with hydrophilic and hydrophobic monomers. The activity of the immobilized enzyme-labeled antibody membranes varied with the thickness of the membranes and increased with decreasing membrane thickness. The activity of the immobilized antibody particles was varied by particle size. Immobilized anti-alpha-fetoprotein particles and membranes can be used for the assay of alpha-fetoprotein by the antigen-antibody reaction, such as a solid-phase sandwich method with high sensitivity.

Kumakura, M.; Kaetsu, I.; Suzuki, M.; Adachi, S.

1983-04-01

277

Radiohalogenated half-antibodies and maleimide intermediate therefor  

DOEpatents

N-(m-radiohalophenyl) maleimide can be conjugated with a reduced antibody having a mercapto group to provide a radiolabelled half-antibody having immunological specific binding characteristics of whole antibody.

Kassis, Amin I. (Chestnut Hill, MA); Khawli, Leslie A. (Newton Centre, MA)

1991-01-01

278

21 CFR 866.3290 - Gonococcal antibody test (GAT).  

Code of Federal Regulations, 2010 CFR

...2010-04-01 false Gonococcal antibody test (GAT). 866.3290 Section 866.3290 Food...866.3290 Gonococcal antibody test (GAT). (a) Identification. A gonococcal antibody test (GAT) is an in vitro device that...

2010-04-01

279

Principles and application of antibody libraries for infectious diseases.  

PubMed

Antibodies have been used efficiently for the treatment and diagnosis of many diseases. Recombinant antibody technology allows the generation of fully human antibodies. Phage display is the gold standard for the production of human antibodies in vitro. To generate monoclonal antibodies by phage display, the generation of antibody libraries is crucial. Antibody libraries are classified according to the source where the antibody gene sequences were obtained. The most useful library for infectious diseases is the immunized library. Immunized libraries would allow better and selective enrichment of antibodies against disease antigens. The antibodies generated from these libraries can be translated for both diagnostic and therapeutic applications. This review focuses on the generation of immunized antibody libraries and the potential applications of the antibodies derived from these libraries. PMID:25214212

Lim, Bee Nar; Tye, Gee Jun; Choong, Yee Siew; Ong, Eugene Boon Beng; Ismail, Asma; Lim, Theam Soon

2014-12-01

280

42 CFR 493.865 - Standard; Antibody identification.  

Code of Federal Regulations, 2010 CFR

... 2010-10-01 false Standard; Antibody identification. 493.865 Section...These Tests § 493.865 Standard; Antibody identification. (a) Failure to... (e) Failure to identify the same antibody in two consecutive or two out of...

2010-10-01

281

Magnetic microparticle antibodies and their application to RIAs  

Microsoft Academic Search

Three types of magnetic microparticle antibodies were developed: 1) magnetic second antibody I (MSA-I) where the antibody molecules were directly immobilized by physical adsorption on Fe3O4 microparticles (magnetic nucleus, MN) 10nm±34% in diameter, 2) magnetic second antibody II (MSA-II) where the antibody molecules were immobilized by chemical coupling on the MN coated with polyacrolein, and 3) magnetic, first antibody (MFA-T3)

Shen Rongsen; Wang Renzhi; Xing Ruiyun; Li Yingoi; Zhou Fengqi; Jiang Shaohua; Lin Zhihao; Xu Banglei

1996-01-01

282

21 CFR 866.5100 - Antinuclear antibody immunological test system.  

...immunochemical techniques the autoimmune antibodies in serum, other...erythematosus (a multisystem autoimmune disease in which antibodies attack the...tissues), hepatitis (a liver disease), rheumatoid arthritis,...

2014-04-01

283

21 CFR 866.5100 - Antinuclear antibody immunological test system.  

Code of Federal Regulations, 2012 CFR

...immunochemical techniques the autoimmune antibodies in serum, other...erythematosus (a multisystem autoimmune disease in which antibodies attack the...tissues), hepatitis (a liver disease), rheumatoid...

2012-04-01

284

21 CFR 866.5100 - Antinuclear antibody immunological test system.  

Code of Federal Regulations, 2011 CFR

...immunochemical techniques the autoimmune antibodies in serum, other...erythematosus (a multisystem autoimmune disease in which antibodies attack the...tissues), hepatitis (a liver disease), rheumatoid...

2011-04-01

285

21 CFR 866.5100 - Antinuclear antibody immunological test system.  

Code of Federal Regulations, 2013 CFR

...immunochemical techniques the autoimmune antibodies in serum, other...erythematosus (a multisystem autoimmune disease in which antibodies attack the...tissues), hepatitis (a liver disease), rheumatoid arthritis,...

2013-04-01

286

21 CFR 866.5100 - Antinuclear antibody immunological test system.  

Code of Federal Regulations, 2010 CFR

...immunochemical techniques the autoimmune antibodies in serum, other...erythematosus (a multisystem autoimmune disease in which antibodies attack the...tissues), hepatitis (a liver disease), rheumatoid...

2010-04-01

287

Progress towards recombinant anti-infective antibodies  

PubMed Central

The global market for monoclonal antibody therapeutics reached a total of $11.2 billion in 2004, with an impressive 42% growth rate over the previous five years and is expected to reach ~$34 billion by 2010. Coupled with this growth are stream-lined product development, production scale-up and regulatory approval processes for the highly conserved antibody structure. While only one of the 21 current FDA-approved antibodies, and one of the 38 products in advanced clinical trials target infectious diseases, there is increasing academic, government and commercial interest in this area. Synagis, an antibody neutralizing respiratory syncitial virus (RSV), garnered impressive sales of $1.1 billion in 2006 in spite of its high cost and undocumented effects on viral titres in human patients. The success of anti-RSV passive immunization has motivated the continued development of anti-infectives to treat a number of other infectious diseases, including those mediated by viruses, toxins and bacterial/fungal cells. Concurrently, advances in antibody technology suggest that cocktails of several monoclonal antibodies with unique epitope specificity or single monoclonal antibodies with broad serotype specificity may be the most successful format. Recent patents and patent applications in these areas will be discussed as predictors of future anti-infective therapeutics. PMID:19149692

Pai, Jennifer C.; Sutherland, Jamie N.; Maynard, Jennifer A.

2009-01-01

288

[Antiganglioside antibodies in neuropathies and motor neuronopathies].  

PubMed

The presence of antiganglioside antibodies is associated with several neurologic disorders. These antibodies recognize several epitopes, generally saccharides present in these glucolipids. The presence of antiGM antibodies has been described in certain clinical syndromes, the main one being multifocal motor neuropathy with and without conduction blocks. The frequency of antiGM1 class IgM antibody falls between 20 and 80% in this disease. Axon predominant Guillain-Barré syndrome is also associated with high titers of antiGM1 antibodies, although in this case class IgG is implicated. The most important association to date has been established between Miller-Fisher syndrome and the presence of antiGQ1b antibodies. Several authors have reported molecular similarities among these gangliosides and bacterial lipopolysaccharides, mainly Campylobacter iejuni. The principal aims in the study of antiganglioside antibodies are to establish their pathogenic role as well as the clinical usefulness of analyzing for them, and to discover new specificities that aid in the diagnosis and classification of neuropathies, whether they are predominantly motor disorders or chronic sensory ones. PMID:9044577

Gallardo, E; Serrano, C; Prat, C; Illa, I

1996-12-01

289

X antigen/antibody markers in hepadnavirus infections. Antibodies to the X gene product(s).  

PubMed

Antibodies to the X antigen of hepatitis B virus and woodchuck hepatitis virus were assayed in serial sera from infected individuals and compared with other markers of infection. Antibody to the X antigen was found in 11 of 17 (65%) patients and 17 of 40 (42%) woodchucks that were surface-antigen positive. In comparison, this antibody was found in 5 of 14 (36%) patients and in none of 4 woodchucks that were surface-antigen negative. In 5 of 6 patients showing seroconversion from hepatitis B e antigen to antibody, antibody to X appeared at or near the time of seroconversion. In patients persistently positive for e antigen, X antibody often appeared when viral DNA became undetectable in the serum. In 14 of 17 (82%) woodchucks positive for antibody to X antigen, it also appeared near or after the time that viral DNA in serum disappeared. X antibodies were detected with great frequency only in populations with high frequencies of other hepatitis B virus markers. The results are consistent with the conclusion that antibody to X antigen is a marker of hepadnavirus infections that seems to be associated with a decrease in viral replication. Antibodies to the X antigen, then, may be a host response to the replication complex of the virus. PMID:2365196

Feitelson, M A; Clayton, M M

1990-08-01

290

Emerging antibody products and Nicotiana manufacturing.  

PubMed

Antibody based products are not widely available to address multiple global health challenges due to high costs, limited manufacturing capacity, and long manufacturing lead times. Nicotiana-based manufacturing of antibody products may now begin to address these challenges as a result of revolutionary advances in transient expression and altered glycosylation pathways. This review provides examples of emerging antibody-based products (mucosal and systemic) that could be competitive and commercially viable when the attributes of Nicotiana-based manufacturing (large scale, versatile, rapid, low cost) are utilized. PMID:21358287

Whaley, Kevin J; Hiatt, Andrew; Zeitlin, Larry

2011-03-01

291

SAbDab: the structural antibody database.  

PubMed

Structural antibody database (SAbDab; http://opig.stats.ox.ac.uk/webapps/sabdab) is an online resource containing all the publicly available antibody structures annotated and presented in a consistent fashion. The data are annotated with several properties including experimental information, gene details, correct heavy and light chain pairings, antigen details and, where available, antibody-antigen binding affinity. The user can select structures, according to these attributes as well as structural properties such as complementarity determining region loop conformation and variable domain orientation. Individual structures, datasets and the complete database can be downloaded. PMID:24214988

Dunbar, James; Krawczyk, Konrad; Leem, Jinwoo; Baker, Terry; Fuchs, Angelika; Georges, Guy; Shi, Jiye; Deane, Charlotte M

2014-01-01

292

Complete Genome Sequence of Treponema pallidum, the  

E-print Network

spirochete, Borrelia burgdorferi, the agent of Lyme disease, identified unique and common genes agent of Lyme disease, are similar in having relatively small genomes and surviv- ing only. The disease quickly reached epidemic pro- portions in Europe and spread across the world during the early 16th

Salzberg, Steven

293

Antigen Specific Plasmacytomas and Antibodies Derived Therefrom.  

National Technical Information Service (NTIS)

Disclosed herein is a method for producing antibodies against an antigen of interest. Animal cells are exposed to both the antigen of interest and a recombinant retroviral vector. The vector contains a combination of oncogenes capable of inducing plasmacy...

R. G. Risser, D. A. Largaespada, J. F. Mushinski, E. M. Wessinger

1990-01-01

294

Detection of typhus antibodies by latex agglutination.  

PubMed Central

A latex test for assay of antibodies to endemic and epidemic typhus rickettsiae is simple, group-specific, sensitive, and reproducible. Cross-reactivity within the typhus group was extensive. PMID:6780601

Hechemy, K E; Osterman, J V; Eisemann, C S; Elliott, L B; Sasowski, S J

1981-01-01

295

[Influence of levamisole on antibody production].  

PubMed

IgM and IgG anti-sheep red blood cells agglutinins were measured from dayy + 2 to day + 30 in SPF mice immunized with 108 SRC and treated with 2.5 or 25 mg LMS/dg. Untreated control mice produced high levels (1:8200) of IgM-antibodies from day + 4 to day + 20. Early, day + 4, IgG-agglutinins appeared together with IgM-antibodies in mice treated with 2.5 mg LMS/kg, whereas administration of 25 mg LMS/kg induced the synthesis of antibodies belonging only to the IgG class. Titres were significantly lower (1:512) in LMS-treated mice than in controls. The switch to IgG of antibody formation is another for the ability of LMS to stimulate T cell activities. PMID:848884

Renoux, G; Renoux, M

1977-01-01

296

Cost modeling for monoclonal antibody manufacturing  

E-print Network

The Novartis BioPharmOps division is responsible for manufacturing large molecule products, including monoclonal antibodies, for late stage clinical trials and commercial sales. The BioPharmOps site in Huningue, France is ...

Simpson, Christina M. (Christina Margaret)

2011-01-01

297

Localization of tumors by radiolabelled antibodies  

Microsoft Academic Search

A method of utilizing radiolabelled antibodies to carcinoembryonic antigens for determining the site of tumors which produce or are associated with carcinoembryonic antigen is disclosed. 3 claims, no drawings.

H. J. Hansen; F. J. Primus

1975-01-01

298

Polynucleotides encoding anti-sulfotyrosine antibodies  

DOEpatents

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.

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

2011-01-11

299

Antibodies against the calcium-binding protein  

SciTech Connect

Plant microsomes contain a protein clearly related to a calcium-binding protein, calsequestrin, originally found in the sarcoplasmic reticulum of muscle cells, responsible for the rapid release and uptake of Ca{sup 2+} within the cells. The location and role of calsequestrin in plant cells is unknown. To generate monoclonal antibodies specific to plant calsequestrin, mice were immunized with a microsomal fraction from cultured cells of Streptanthus tortuosus (Brassicaceae). Two clones cross-reacted with one protein band with a molecular weight equal to that of calsequestrin (57 kilodaltons) by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting. This band is able to bind {sup 45}Ca{sup 2+} and can be recognized by a polyclonal antibody against the canine cardiac muscle calsequestrin. Rabbit skeletal muscle calsequestrin cross-reacted with the plant monoclonal antibodies. The plant monoclonal antibodies generated here are specific to calsequestrin protein.

Chou, Mei; Jensen, K.G.; Sjolund, R.D. (Univ. of Iowa, Iowa City (USA)); Krause, K.H.; Campbell, K.P. (Univ. of Iowa College of Medicine, Iowa City (USA))

1989-12-01

300

Analysis of antibody assay methods and classes of viral antibodies in serodiagnosis of cytomegalovirus infection.  

PubMed Central

Forty-nine serum pairs with antibody to cytomegalovirus (CMV) were evaluated for rises in antibody titer (greater than or equal to fourfold) by indirect hemagglutination (IHA) and complement fixation (CF), using a freeze-thaw antigen (FT) and a glycine extract antigen (GE). In this sample CF-FT detected more rises in antibody titer than did CF-GE. IHA detected the least number. The apparent reason for stationary antibody titers with CF-GE and IHA was the presence of high antibody titers in the first serum specimen. Separation of immunoglobulin classes of 20 serum pairs by sucrose gradient centrifugation indicated that these antibodies with IHA were of the immunoglobulin M (IgM) class and those with CF-GE were of the IgG class. By separation of immunoglobulin classes, rises in IgG CMV antibody titers were seen with IHA, rises not observed in the whole serum because of high IgM antibody titers in the first serum specimen. Absence of rises in antibody titers with CF-FT was due in part to too early sampling of the second serum specimen (less than 21 days) and in part to an apparent inability of some individuals to respond with antibody reactive with FT antigen. CF-GE and CF-FT antibodies of the IgM class were detected in some sera, usually in specimens collected more than 10 days after the onset of symptoms. Although reactive with CMV antigen, the specificity of these IgM antibodies in relation to rheumatoid factor requires clarification. PMID:212446

Cremer, N E; Hoffman, M; Lennette, E H

1978-01-01

301

454 antibody sequencing - error characterization and correction  

PubMed Central

Background 454 sequencing is currently the method of choice for sequencing of antibody repertoires and libraries containing large numbers (106 to 1012) of different molecules with similar frameworks and variable regions which poses significant challenges for identifying sequencing errors. Identification and correction of sequencing errors in such mixtures is especially important for the exploration of complex maturation pathways and identification of putative germline predecessors of highly somatically mutated antibodies. To quantify and correct errors incorporated in 454 antibody sequencing, we sequenced six antibodies at different known concentrations twice over and compared them with the corresponding known sequences as determined by standard Sanger sequencing. Results We found that 454 antibody sequencing could lead to approximately 20% incorrect reads due to insertions that were mostly found at shorter homopolymer regions of 2-3 nucleotide length, and less so by insertions, deletions and other variants at random sites. Correction of errors might reduce this population of erroneous reads down to 5-10%. However, there are a certain number of errors accounting for 4-8% of the total reads that could not be corrected unless several repeated sequencing is performed, although this may not be possible for large diverse libraries and repertoires including complete sets of antibodies (antibodyomes). Conclusions The experimental test procedure carried out for assessing 454 antibody sequencing errors reveals high (up to 20%) incorrect reads; the errors can be reduced down to 5-10% but not less which suggests the use of caution to avoid false discovery of antibody variants and diversity. PMID:21992227

2011-01-01

302

Construction of human antibody gene libraries and selection of antibodies by phage display.  

PubMed

Antibody phage display is the most commonly used in vitro selection technology and has yielded thousands of useful antibodies for research, diagnostics, and therapy.The prerequisite for successful generation and development of human recombinant antibodies using phage display is the construction of a high-quality antibody gene library. Here, we describe the methods for the construction of human immune and naive scFv gene libraries.The success also depends on the panning strategy for the selection of binders from these libraries. In this article, we describe a panning strategy that is high-throughput compatible and allows parallel selection in microtiter plates. PMID:24037844

Frenzel, André; Kügler, Jonas; Wilke, Sonja; Schirrmann, Thomas; Hust, Michael

2014-01-01

303

Antibody Responses during Hepatitis B Viral Infection  

PubMed Central

Hepatitis B is a DNA virus that infects liver cells and can cause both acute and chronic disease. It is believed that both viral and host factors are responsible for determining whether the infection is cleared or becomes chronic. Here we investigate the mechanism of protection by developing a mathematical model of the antibody response following hepatitis B virus (HBV) infection. We fitted the model to data from seven infected adults identified during acute infection and determined the ability of the virus to escape neutralization through overproduction of non-infectious subviral particles, which have HBs proteins on their surface, but do not contain nucleocapsid protein and viral nucleic acids. We showed that viral clearance can be achieved for high anti-HBV antibody levels, as in vaccinated individuals, when: (1) the rate of synthesis of hepatitis B subviral particles is slow; (2) the rate of synthesis of hepatitis B subviral particles is high but either anti-HBV antibody production is fast, the antibody affinity is high, or the levels of pre-existent HBV-specific antibody at the time of infection are high, as could be attained by vaccination. We further showed that viral clearance can be achieved for low equilibrium anti-HBV antibody levels, as in unvaccinated individuals, when a strong cellular immune response controls early infection. PMID:25078553

Ciupe, Stanca M.; Ribeiro, Ruy M.; Perelson, Alan S.

2014-01-01

304

Decay of maternal antibodies in broiler chickens.  

PubMed

The objective of this study was to determine the decay rate of maternal antibodies against major broiler chicken pathogens. A total of 30 one-day-old broiler chicks were obtained from a commercial hatchery and reared in isolation. These chicks were retrieved from a parent flock that received a routine vaccination program. Chicks were bled at hatch and sequentially thereafter every 5 d through 30 d of age. Maternal antibody titers were measured by ELISA for avian encephalomyelitis (AEV), avian influenza virus (AIV), chicken anemia virus (CAV), infectious bursal disease virus (IBDV), infectious bronchitis virus (IBV), infectious laryngotracheitis virus (ILTV), Mycoplasma gallisepticum (MG), Mycoplasma synoviae (MS), and reovirus (Reo). Maternal antibody titers for Newcastle disease virus (NDV) were measured using a hemagglutination inhibition test. Half-life estimates of maternal antibody titers were 5.3, 4.2, 7, 5.1, 3.9, 3.8, 4.9, 4.1, 6.3, and 4.7 d for AEV, AIV, CAV, IBDV, IBV, ILTV, MG, MS, NDV, and Reo, respectively. The statistical analysis revealed significant differences among half-lives of maternal antibody titers against certain pathogens. Furthermore, all maternal antibody titers were depleted by 10 d of age except for IBDV. PMID:23960115

Gharaibeh, Saad; Mahmoud, Kamel

2013-09-01

305

Comparison of fluorescent-antibody, neutralizing-antibody, and complement-enhanced neutralizing-antibody assays for detection of serum antibody to respiratory syncytial virus.  

PubMed

A comparison of three assays for the detection of serum antibody to respiratory syncytial virus (RSV) was carried out on 47 serum samples obtained sequentially from infants and young children with RSV infection. Neutralizing-antibody (NA) activity was determined by a semimicromethod of plaque reduction. Complement-enhanced NA activity was determined by the addition of guinea pig complement to NA assays. RSV antibody responses in immunoglobulin G, immunoglobulin M, and immunoglobulin A classes were determined by using indirect immunofluorescence techniques for fluorescent-antibody (FAb) assay. Antibody to RSV was detectable by all three techniques as early as 4 days after the onset of illness. At all phases of illness, titers obtained by complement-enhanced NA assays were significantly greater than those obtained by NA or FAb assays (P less than 0.01). RSV-FAb titers determined in the immunoglobulin G class correlated well with those determined by complement-enhanced NA or NA assays. The data suggest that the FAb assay for detection of RSV antibody in serum is somewhat less sensitive but also less laborious and more rapid than NA assays. PMID:7016915

Kaul, T N; Welliver, R C; Ogra, P L

1981-05-01

306

Fetal arthrogryposis and maternal serum antibodies.  

PubMed

Arthrogryposis multiplex congenital (AMC) describes multiple joint contractures resulting from lack of movement in utero. Antibodies directed at the fetal isoform of the muscle acetylcholine receptor (AChR) have been reported in a small number of asymptomatic mothers of AMC babies. We examined sera from 179 mothers of AMC babies and 20 parous and non-parous controls to look for antibodies to AChR or undefined muscle or neuronal proteins. We found positive AChR antibodies in only three sera (1.5%) from asymptomatic AMC mothers. However, there was reactivity with muscle or with neuronal antigens in 33% of the sera, and reactivity to undefined neuronal antigens was more common in sera from mothers of AMC babies with CNS involvement (p=0.001) than those without. The offspring of mothers with AChR antibodies may benefit from treatment during pregnancy. Other maternal antibodies require further study, but these observations add to the emerging literature on maternal antibodies associated with developmental intrauterine disorders. PMID:16919948

Dalton, Paola; Clover, Linda; Wallerstein, Robert; Stewart, Helen; Genzel-Boroviczeny, Orsolya; Dean, Andrew; Vincent, Angela

2006-08-01

307

Avian Diagnostic and Therapeutic Antibodies to Viral Emerging Pathogens  

SciTech Connect

During the current period the following key objectives were achieved: demonstration of high titer antibody production by geese following immunization with inactived H1N1 virus; completion of the epitope mapping of West Nile Virus-specific goose antibodies and initiation of epitope mapping of H1N1 flu-specific goose antibodies; advancement in scalable purification of goose antibodies.

David Bradley

2011-03-31

308

Mechanisms of Allergen-Antibody Interaction of Cockroach Allergen Bla g 2 with Monoclonal Antibodies  

E-print Network

Mechanisms of Allergen-Antibody Interaction of Cockroach Allergen Bla g 2 with Monoclonal is strongly associated with asthma, and involves the production of IgE antibodies against inhaled allergens. Reports of conformational epitopes on inhaled allergens are limited. The conformational epitopes for two

309

High level transient production of recombinant antibodies and antibody fusion proteins in HEK293 cells  

PubMed Central

Background The demand of monospecific high affinity binding reagents, particularly monoclonal antibodies, has been steadily increasing over the last years. Enhanced throughput of antibody generation has been addressed by optimizing in vitro selection using phage display which moved the major bottleneck to the production and purification of recombinant antibodies in an end-user friendly format. Single chain (sc)Fv antibody fragments require additional tags for detection and are not as suitable as immunoglobulins (Ig)G in many immunoassays. In contrast, the bivalent scFv-Fc antibody format shares many properties with IgG and has a very high application compatibility. Results In this study transient expression of scFv-Fc antibodies in human embryonic kidney (HEK) 293 cells was optimized. Production levels of 10-20 mg/L scFv-Fc antibody were achieved in adherent HEK293T cells. Employment of HEK293-6E suspension cells expressing a truncated variant of the Epstein Barr virus (EBV) nuclear antigen (EBNA) 1 in combination with production under serum free conditions increased the volumetric yield up to 10-fold to more than 140 mg/L scFv-Fc antibody. After vector optimization and process optimization the yield of an scFv-Fc antibody and a cytotoxic antibody-RNase fusion protein further increased 3-4-fold to more than 450 mg/L. Finally, an entirely new mammalian expression vector was constructed for single step in frame cloning of scFv genes from antibody phage display libraries. Transient expression of more than 20 different scFv-Fc antibodies resulted in volumetric yields of up to 600 mg/L and 400 mg/L in average. Conclusion Transient production of recombinant scFv-Fc antibodies in HEK293-6E in combination with optimized vectors and fed batch shake flasks cultivation is efficient and robust, and integrates well into a high-throughput recombinant antibody generation pipeline. PMID:23802841

2013-01-01

310

IgG2 antibodies block IgE antibody-induced asthma in guinea pigs.  

PubMed

In order to examine the blocking activity of IgG2 antibodies to guinea pig for IgE antibodies-induced guinea pig asthma, experiments were carried out as follows. Guinea pigs were passively sensitized intravenously with guinea pig serum containing IgE antibodies to ovalbumin (OA). 8 days after sensitization, IgG2 purified from guinea pigs hyperimmunized with OA was intravenously injected. One hour later, the guinea pigs were challenged by inhalation of OA solution. Asthma attacks were not observed in the guinea pigs, whereas the attacks were observed in guinea pigs passively sensitized with the IgE antibodies but injected IgG2 fraction from normal guinea pigs 1 h before inhalation. These observations suggested that IgG antibodies that increased after immunotherapy might block asthma caused by inhalation of allergens in humans. PMID:3957447

Yamauchi, N; Ito, K; Suko, M; Ishii, A; Miyamoto, T

1986-01-01

311

Antibody phage display libraries: contributions to oncology.  

PubMed

Since the advent of phage display technology, dating back to 1985, antibody libraries displayed on filamentous phage surfaces have been used to identify specific binders for many different purposes, including the recognition of tumors. Phage display represents a high-throughput technique for screening billions of random fusion antibodies against virtually any target on the surface or inside cancer cells, or even soluble markers found in patient serum. Many phage display derived binders targeting important tumor markers have been identified. Selection directed to tumoral cells' surfaces lead to the identification of unknown tumoral markers. Also the improvement of methods that require smaller amounts of cells has opened the possibility to use this approach on patient samples. Robust techniques combining an antibody library displayed on the phage surface and protein microarray allowed the identification of auto antibodies recognized by patient sera. Many Ab molecules directly or indirectly targeting angiogenesis have been identified, and one of them, ramucirumab, has been tested in 27 phase I-III clinical trials in a broad array of cancers. Examples of such antibodies will be discussed here with emphasis on those used as probes for molecular imaging and other clinical trials. PMID:22754305

Dantas-Barbosa, Carmela; de Macedo Brigido, Marcelo; Maranhao, Andrea Queiroz

2012-01-01

312

[Therapeutic monoclonal antibodies in onco-hematology].  

PubMed

Rituximab, a chimeric monoclonal anti-CD20 antibody, was introduced into clinical practice in 1997, and has proven to be highly effective in the treatment of B-lymphoproliferative disorders and autoimmune diseases. Despite such success, in vivo mechanisms of action of anti-CD20 have only recently began to be unraveled, pointing to the crucial role of antibody-dependent cellular cytotoxicity response mediated through Fcg receptor signalling. Better understanding of pharmacokinetics and factors influencing individual exposure mediated through anti-CD20 will allow to engineer these molecules to increase their effector responses. Meanwhile, other formats have also been investigated, such as radiolabeled anti-CD20, or coupling of antibodies to cytotoxic drugs such as anti-CD33 used in myeloid leukemia. However these antibodies are used in combination with standard chemotherapy and cannot substitute for cytotoxic drugs. This review summarizes the knowledge acquired through our clinical use of anti-CD20 and authorized monoclonal antibodies in oncohematology and proposes some news areas that will lead to the development of new and more effective therapeutic strategies. PMID:20035683

Cartron, Guillaume; Rossi, Jean-François

2009-12-01

313

Antibodies designed as effective cancer vaccines  

PubMed Central

Antigen/antibody complexes can efficiently target antigen presenting cells to allow stimulation of the cellular immune response. Due to the difficulty of manufacture and their inherent instability complexes have proved inefficient cancer vaccines. However, anti-idiotypic antibodies mimicking antigens have been shown to stimulate both antibody and T cell responses. The latter are due to T cell mimotopes expressed within the complementarity-determining regions (CDRs) of antibodies that are efficiently presented to dendritic cells in vivo. Based on this observation we have designed a DNA vaccine platform called ImmunoBody™, where cytotoxic T lymphocyte (CTL) and helper T cell epitopes replace CDR regions within the framework of a human IgG1 antibody. The ImmunoBody™ expression system has a number of design features which allow for rapid production of a wide range of vaccines. The CDR regions of the heavy and light chain have been engineered to contain unique restriction endonuclease sites, which can be easily opened, and oligonucleotides encoding the T cell epitopes inserted. The variable and constant regions of the ImmunoBody™ are also flanked by restriction sites, which permit easy exchange of other IgG subtypes. Here we show a range of T cell epitopes can be inserted into the ImmunoBody™ vector and upon immunization these T cell epitopes are efficiently processed and presented to stimulate high frequency helper and CTL responses capable of anti-tumor activity. PMID:20046577

Metheringham, RL; Pudney, VA; Gunn, B; Towey, M; Spendlove, I

2009-01-01

314

[Prevalence of antinuclear envelope antibodies and their isotypes in sera positive for antinuclear antibodies].  

PubMed

Antinuclear antibodies detected in HEp-2 cells by indirect immunofluorescence assay display a great variety of images, including the nuclear envelope pattern. This is quite a less frequent finding. Two thousand five hundred and ninety-four sera were processed, and 37.6% of ANA were detected. The prevalence of anti-nuclear envelope antibodies (ANEA) was of 1.2%, with a high association with autoimmune liver diseases (83%) and a low association with systemic lupus erythematosus. In 21 sera of patients with ANEA, no anti-DNAn antibodies were found; but 28.6% of anti-smooth muscle antibodies and 19% of anti-mitochondrial antibodies were detected. The triple rodent tissue section proved to be a less sensitive substrate than HEp-2 for the detection of ANEA. When using conjugates against different isotypes of antibodies for the detection of ANA, 90.5% of IgG, 66.6% of IgA and 9.5% of IgM. Two patients had ANEA-IgA at high titers (> or = 1:160) without ANEA-IgG. In this work, the importance of performing complementary tests for the detection of anti-smooth muscle antibodies, anti-mitochondrial antibodies and anti-DNAn is highlighted in order to apply these tests as guidelines for the clinical diagnosis of patients with ANEA. Besides, this study expresses the need of using total anti-Ig antibodies as conjugate for IIF-HEp-2 instead of anti-lgG; until the role of IgA antibodies in these autoimmune diseases is clarified. PMID:16977968

Arcavi, Miriam; Orfus, Gladys

2006-01-01

315

Back to the future: recombinant polyclonal antibody therapeutics  

PubMed Central

Antibody therapeutics are one of the fastest growing classes of pharmaceuticals, with an annual US market over $20 billion, developed to treat a variety of diseases including cancer, auto-immune and infectious diseases. Most are currently administered as a single molecule to treat a single disease, however there is mounting evidence that cocktails of multiple antibodies, each with a unique binding specificity and protective mechanism, may improve clinical efficacy. Here, we review progress in the development of oligoclonal combinations of antibodies to treat disease, focusing on identification of synergistic antibodies. We then discuss the application of modern antibody engineering technologies to produce highly potent antibody preparations, including oligoclonal antibody cocktails and truly recombinant polyclonal antibodies. Specific examples illustrating the synergy conferred by multiple antibodies will be provided for diseases caused by botulinum toxin, cancer and immune thrombocytopenia. The bioprocessing and regulatory options for these preparations will be discussed. PMID:24443710

Wang, Xian-zhe; Coljee, Vincent W.; Maynard, Jennifer A.

2013-01-01

316

The maturation of antibody technology for the HIV epidemic.  

PubMed

Antibodies are one of our most useful biological tools. Indeed, improvements in antibody-based technologies have ushered in a new era of antibody-based therapeutics, research and diagnostic tools. Although improved technologies have led to the development of therapeutic antibodies for treatment of malignancies and inflammatory conditions, the use of advanced antibody technology in the therapy of viral infections is in its infancy. Non-human primate studies have demonstrated that antibodies against the HIV envelope can both prevent viral infection and control viremia. Despite the obvious potential of antibody therapies against HIV, there remain limitations in production and purification capacity that require further research. Recent advances in recombinant antibody technology have led to the development of a range of novel antibody fragments, such as single-domain nanobodies and bispecific antibodies, that are capable of targeting cancer cells to cytotoxic T cells. Novel antibody production techniques have also been designed, allowing antibodies to be obtained from non-mammalian cells, bovine colostrum and the periplasm and cytoplasm of bacteria. These advances may allow large-scale production of HIV antibodies that are capable of protecting against HIV infection or serving as therapeutics that reduce the need for life-long antiretroviral treatment. This review summarises recent advances in antibody-based technologies and discusses the possibilities and challenges of using these advances to design prophylactics and therapeutics against HIV. PMID:24797582

Winnall, Wendy R; Beasley, Matthew D; Center, Rob J; Parsons, Matthew S; Kiefel, Ben R; Kent, Stephen J

2014-08-01

317

Removal of Species Constraints in Antibody Detection ?  

PubMed Central

Serum antibodies from myriad species, particularly birds, can provide key information regarding the transmission and the expansion of the territory of emerging pathogens. Expedient antibody analysis is constrained by a lack of species-specific reagents, a deficiency potentially highlighted by the recent swine-origin influenza A virus (H1N1) outbreak. Available methodologies present difficulties that discourage thorough serologic monitoring of potential disease vectors or hosts. Rapid high-throughput procedures that combined serum amine labeling via biotinylation, contaminant removal, and microsphere-based immunoassays for antibodies to three arboviruses were developed. Agent-specific adaptations of this simple format should facilitate expanded surveillance and diagnostic capabilities regarding pathogens of human and veterinary importance. PMID:19923570

Basile, Alison Jane; Biggerstaff, Brad J.; Kosoy, Olga L.; Junna, Shilpa R.; Panella, Nicholas A.; Powers, Ann M.; Stark, Lillian M.; Nemeth, Nicole M.

2010-01-01

318

Method for altering antibody light chain interactions  

DOEpatents

A method for recombinant antibody subunit dimerization including modifying at least one codon of a nucleic acid sequence to replace an amino acid occurring naturally in the antibody with a charged amino acid at a position in the interface segment of the light polypeptide variable region, the charged amino acid having a first polarity; and modifying at least one codon of the nucleic acid sequence to replace an amino acid occurring naturally in the antibody with a charged amino acid at a position in an interface segment of the heavy polypeptide variable region corresponding to a position in the light polypeptide variable region, the charged amino acid having a second polarity opposite the first polarity. Nucleic acid sequences which code for novel light chain proteins, the latter of which are used in conjunction with the inventive method, are also provided.

Stevens, Fred J. (Naperville, IL); Stevens, Priscilla Wilkins (Evanston, IL); Raffen, Rosemarie (Elmhurst, IL); Schiffer, Marianne (Downers Grove, IL)

2002-01-01

319

Losing your nerves? Maybe it's the antibodies  

PubMed Central

We propose that the normal immunocompetent B cell repertoire is replete with B cells making antibodies that recognize brain antigens. Although B cells that are reactive with self antigen are normally silenced during B cell maturation, the blood–brain barrier (BBB) prevents many brain antigens from participating in this process. This enables the generation of a B cell repertoire that is sufficiently diverse to cope with numerous environmental challenges. It requires, however, that the integrity of the BBBs is uninterrupted throughout life to protect the brain from antibodies that crossreact with microorganisms and brain antigens. Under conditions of BBB compromise, and during fetal development, we think that these antibodies can alter brain function in otherwise healthy individuals. PMID:19424277

Diamond, Betty; Huerta, Patricio T.; Mina-Osorio, Paola; Kowal, Czeslawa; Volpe, Bruce T.

2009-01-01

320

Monoclonal antibodies specific for sickle cell hemoglobin  

SciTech Connect

Two mouse hybridoma cell lines were isolated which produce monoclonal antibodies that bind hemoglobin S. The mice were immunized with peptide-protein conjugates to stimulate a response to the amino terminal peptide of the beta chain of hemoglobin S, where the single amino acid difference between A and S occurs. Immunocharacterization of the antibodies shows that they bind specifically to the immunogen peptide and to hemoglobin S. The specificity for S is high enough that one AS cell in a mixture with a million AA cells is labeled by antibody, and such cells can be analyzed by flow cytometry. Immunoblotting of electrophoretic gels allows definitive identification of hemoglobin S as compared with other hemoglobins with similar electrophoretic mobility. 12 references, 4 figures.

Jensen, R.H.; Vanderlaan, M.; Grabske, R.J.; Branscomb, E.W.; Bigbee, W.L.; Stanker, L.H.

1985-01-01

321

Method for preparation of single chain antibodies  

DOEpatents

This invention provides a method for identifying cells expressing a target single chain antibody (scFv) directed against a target antigen from a collection of cells that includes cells that do not express the target scFv, comprising the step of combining the collection of cells with an anti-idiotype directed to an antibody specific for the target antigen and detecting interaction, if any, of the anti-idiotype with the cells, wherein the occurrence of an interaction identifies the cell as one which expresses the target scFv. This invention also provides a method for making a single chain antibody (scFv) directed against an antigen, wherein the selection of clones is made based upon interaction of those clones with an appropriate anti-idiotype, and heretofore inaccessible scFv so made. This invention provides the above methods or any combination thereof. Finally, this invention provides various uses of these methods.

Cheung, Nai-Kong V. (New York, NY); Guo, Hong-fen (New York, NY)

2012-04-03

322

Antibody-based biological toxin detection  

SciTech Connect

Fiber optic evanescent fluorosensors are under investigation in our laboratory for the study of drug-receptor interactions for detection of threat agents and antibody-antigen interactions for detection of biological toxins. In a direct competition assay, antibodies against Cholera toxin, Staphylococcus Enterotoxin B or ricin were noncovalently immobilized on quartz fibers and probed with fluorescein isothiocyanate (FITC) - labeled toxins. In the indirect competition assay, Cholera toxin or Botulinum toxoid A was immobilized onto the fiber, followed by incubation in an antiserum or partially purified anti-toxin IgG. These were then probed with FITC-anti-IgG antibodies. Unlabeled toxins competed with labeled toxins or anti-toxin IgG in a dose dependent manner and the detection of the toxins was in the nanomolar range.

Menking, D.E.; Goode, M.T. [Army Edgewood Research, Development and Engineering Center, Aberdeen Proving Ground, MD (United States)

1995-12-01

323

In vivo modulators of antibody kinetics.  

PubMed

The aim of the present study was to summarize the effect of in vivo modulation of antibody kinetics and to present new data on the in vivo effect of the cell membrane active detergent Tween 80 and the cytokine interleukin-2 (IL-2) on the accumulation and clearance of a radioactive antibody. Mice bearing Lewis lung carcinoma xenografts and rats bearing DMBA-induced mammary carcinomas were studied after injecting I-125 labeled IgG1 monoclonal antibody (3c4c7g6) raised against a tyrosine kinase receptor protein Tie. Expression of Tie is known to be abundant in vascular endothelia and possibly related to malignant angiogenesis. Tween 80 was administered intratumorally (0.04% of tumor volume), whereas IL-2 was administered intraperitoneally. In the Lewis lung tumor model, the absolute tumor uptake varied between 2 and 5% ID/g, and maximum uptake was achieved after 24 h with Tween, and after 48 h without Tween. Tween manipulation did not increase the uptake in any normal organ, but it enhanced antibody clearance from the blood. In the DMBA rat model, IL-2 had no effect on blood clearance, but enhanced the uptake of Tie antibody into the tumor from 2.5-0.9 to 4.5-0.4% ID/g at 48 h. These data indicate that antibody biodistribution and pharmacokinetics can be modulated by a surface detergent and a cytokine, giving decreased exposure to critical organs, and increased uptake into the tumor. This type of manipulation provides an opportunity to optimize radioimmunotherapy. PMID:8679255

Jekunen, A; Kairemo, K; Karnani, P

1996-01-01

324

Production and characterization of antibodies against microcystins.  

PubMed

Antibodies against a microcystin (MCYST) leucine-arginine variant (MCYST-LR) were demonstrated 4 weeks after immunization of rabbits with either MCYST-LR-polylysine- or MCYST-LR-ethylenediamine-modified bovine serum albumin. A radioimmunoassay (RIA), a direct competitive enzyme-linked immunosorbent assay (ELISA), and an indirect competitive ELISA were developed for characterization of the antibodies. Indirect ELISA and RIA revealed that MCYST-LR-ethylenediamine-bovine serum albumin was a better immunogen. Competitive RIA and direct ELISA revealed that the antibodies had good cross-reactivities with an MCYST-arginine-arginine variant (MCYST-RR), MCYST-LR, an MCYST-tyrosine-arginine variant (MCYST-YR), and nodularin (NODLN); but they had lower reactivities with variants MCYST-leucine-tyrosine (MCYST-LY) and MCYST-leucine-alanine (MCYST-LA). The antibodies did not cross-react with ozonolyzed MCYST-LR. The concentrations causing 50% inhibition of binding of reduced MCYST-LR to the antibodies by MCYST-RR, MCYST-LR, MCYST-YR, NODLN, MCYST-LA, and MCYST-LY in the RIA were 43, 105, 112, 503, 671, and 1,920 ng/ml, respectively. The concentrations causing 50% inhibition of binding of MCYST-LR-horseradish peroxidase to the antibodies by MCYST-RR, MCYST-LR, MCYST-YR, NODLN, MCYST-LY, and MCYST-LA in the ELISA were 1.75, 2.2, 3.4, 4.6, 50, and 114 ng/ml, respectively. PMID:2506810

Chu, F S; Huang, X; Wei, R D; Carmichael, W W

1989-08-01

325

Production and characterization of antibodies against microcystins.  

PubMed Central

Antibodies against a microcystin (MCYST) leucine-arginine variant (MCYST-LR) were demonstrated 4 weeks after immunization of rabbits with either MCYST-LR-polylysine- or MCYST-LR-ethylenediamine-modified bovine serum albumin. A radioimmunoassay (RIA), a direct competitive enzyme-linked immunosorbent assay (ELISA), and an indirect competitive ELISA were developed for characterization of the antibodies. Indirect ELISA and RIA revealed that MCYST-LR-ethylenediamine-bovine serum albumin was a better immunogen. Competitive RIA and direct ELISA revealed that the antibodies had good cross-reactivities with an MCYST-arginine-arginine variant (MCYST-RR), MCYST-LR, an MCYST-tyrosine-arginine variant (MCYST-YR), and nodularin (NODLN); but they had lower reactivities with variants MCYST-leucine-tyrosine (MCYST-LY) and MCYST-leucine-alanine (MCYST-LA). The antibodies did not cross-react with ozonolyzed MCYST-LR. The concentrations causing 50% inhibition of binding of reduced MCYST-LR to the antibodies by MCYST-RR, MCYST-LR, MCYST-YR, NODLN, MCYST-LA, and MCYST-LY in the RIA were 43, 105, 112, 503, 671, and 1,920 ng/ml, respectively. The concentrations causing 50% inhibition of binding of MCYST-LR-horseradish peroxidase to the antibodies by MCYST-RR, MCYST-LR, MCYST-YR, NODLN, MCYST-LY, and MCYST-LA in the ELISA were 1.75, 2.2, 3.4, 4.6, 50, and 114 ng/ml, respectively. PMID:2506810

Chu, F S; Huang, X; Wei, R D; Carmichael, W W

1989-01-01

326

IgA antibodies in Rett syndrome.  

PubMed

The level of IgA antibodies to gluten and gliadin proteins found in grains and to casein found in milk, as well as the level of IgG to gluten and gliadin, have been examined in 23 girls with Rett syndrome and 53 controls. Highly statistically significant increases were found for the Rett population compared to the controls. The reason for this remains unknown, but because IgA antibodies reflect the uptake of proteins and/or epitopes of proteins from the gut, this may be indicative of increased protein uptake. PMID:16613867

Reichelt, K L; Skjeldal, O

2006-03-01

327

Antibodies for treatment of Clostridium difficile infection.  

PubMed

Antibodies for the treatment of Clostridium difficile infection (CDI) have been demonstrated to be effective in the research and clinical environments. Early uncertainties about molecular and treatment modalities now appear to have converged upon the systemic dosing of mixtures of human IgG1. Although multiple examples of high-potency monoclonal antibodies (MAbs) exist, significant difficulties were initially encountered in their discovery. This minireview describes historical and contemporary MAbs and highlights differences between the most potent MAbs, which may offer insight into the pathogenesis and treatment of CDI. PMID:24789799

Humphreys, David P; Wilcox, Mark H

2014-07-01

328

Natural killer cell mediated antibody-dependent cellular cytotoxicity in tumor immunotherapy with therapeutic antibodies.  

PubMed

In the last decade several therapeutic antibodies have been Federal Drug Administration (FDA) and European Medicines Agency (EMEA) approved. Although their mechanisms of action in vivo is not fully elucidated, antibody-dependent cellular cytotoxicity (ADCC) mediated by natural killer (NK) cells is presumed to be a key effector function. A substantial role of ADCC has been demonstrated in vitro and in mouse tumor models. However, a direct in vivo effect of ADCC in tumor reactivity in humans remains to be shown. Several studies revealed a predictive value of Fc?RIIIa-V158F polymorphism in monoclonal antibody treatment, indicating a potential effect of ADCC on outcome for certain indications. Furthermore, the use of therapeutic antibodies after allogeneic hematopoietic stem cell transplantation is an interesting option. Studying the role of the Fc?RIIIa-V158F polymorphism and the influence of Killer-cell Immunoglobuline-like Receptor (KIR) receptor ligand incompatibility on ADCC in this approach may contribute to future transplantation strategies. Despite the success of approved second-generation antibodies in the treatment of several malignancies, efforts are made to further augment ADCC in vivo by antibody engineering. Here, we review currently used therapeutic antibodies for which ADCC has been suggested as effector function. PMID:23543707

Seidel, Ursula J E; Schlegel, Patrick; Lang, Peter

2013-01-01

329

Increasing stability of antibody via antibody engineering: Stability engineering on an anti-hVEGF.  

PubMed

Antibody stability is very important for expression, activity, specificity, and storage. This knowledge of antibody structure has made it possible for a computer-aided molecule design to be used to optimize and increase antibody stability. Many computational methods have been built based on knowledge or structure, however, a good integrated engineering system has yet to be developed that combines these methods. In the current study, we designed an integrated computer-aided engineering protocol, which included several successful methods. Mutants were designed considering factors that affected stability and multiwall filter screening was used to improve the design accuracy. Using this protocol, the thermo-stability of an anti-hVEGF antibody was significantly improved. Nearly 40% of the single-point mutants proved to be more stable than the parent antibody and most of the mutations could be stacked effectively. The T50 also improved about 7°C by combinational mutation of seven sites in the light chain and three sites in the heavy chain. Data indicate that the protocol is an effective method for optimization of antibody structure, especially for improving thermo-stability. This protocol could also be used to enhance the stability of other antibodies. Proteins 2014; 82:2620-2630. © 2014 Wiley Periodicals, Inc. PMID:24916692

Wang, Shuang; Liu, Ming; Zeng, Dadi; Qiu, Weiyi; Ma, Pingping; Yu, Yunzhou; Chang, Hongyan; Sun, Zhiwei

2014-10-01

330

Antibody inhibition of protein activity in starfish oocytes.  

PubMed

Antibodies are widely utilized in cell and molecule biology for immunoblots, immunostaining, immunoprecipitation, immunoaffinity purification, and immunoassay. Some antibodies can be used for in vivo inhibition experiments. These antibodies bind to their target molecules and neutralize their functions, providing functional information in the study of their biological role. Here, we describe our methods for obtaining inhibitory antibodies against desired proteins. We then describe in the starfish oocyte system how to inhibit a target protein, even in the nucleus, by injection of antibody into the cytoplasm, and how to evaluate antibody inhibition of cell cycle regulators in small numbers of oocytes. PMID:24567224

Okumura, Eiichi; Hara, Masatoshi; Kishimoto, Takeo

2014-01-01

331

Novel human antibody therapeutics: The age of the Umabs  

PubMed Central

Monoclonal antibodies represent a major and increasingly important category of biotechnology products for the treatment of human diseases. The state-of-the-art of antibody technology has evolved to the point where therapeutic monoclonal antibodies, that are practically indistinguishable from antibodies induced in humans, are routinely generated. We depict how our science-based approach can be used to further improve the efficacy of antibody therapeutics, illustrated by the development of three monoclonal antibodies for various cancer indications: zanolimumab (directed against CD4), ofatumumab (directed against CD20) and zalutumumab (directed against epidermal growth factor receptor). PMID:18702090

Ruuls, Sigrid R; van Bueren, Jeroen J Lammerts; van de Winkel, Jan G J; Parren, Paul W H I

2008-01-01

332

Equine antihapten antibody. The subunits and fragments of anti-beta-lactoside antibody.  

PubMed

Eight antigenically unique immunoglobulins have been identified in purified equine anti-p-azophenyl-beta-lactoside (Lac) antibody isolated from a single horse. The Fc fragments of the gammaGa-, gammaGb-, gammaGc-, and -gammaA-globulins have been shown to possess unique antigenic determinants. Common gammaG- and gammaA-Fc fragment antigenic determinants, which were absent from the 10Sgamma(1)- and gammaM-globulins, have also been observed. All antibody populations share two antigenically distinct light (B, L) chain variants. The association of anti-Lac antibody with the hapten p-(p-dimethylamino-benzeneazo)-phenyl-beta-lactoside has been measured by equilibrium dialysis and by fluorescence quenching. A variation in the affinity of anti-Lac antibody for hapten has been observed. The affinity of antibody was unaltered by enzymatic removal of the Fc fragments by peptic digestion or dissociation of the two combining sites on the papain 3.5S Fab fragments, indicating that the observed heterogeneity of affinities was not a direct function of the heterogeneity in structure of the Fc fragments. Isolated heavy (A, H) chains of gammaA-anti-Lac antibody have been shown to have retamed affinity for Lac dye by equilibrium dialysis and by analytical ultracentrifugation, employing a combination of schlieren and absorption optics. The heavy (A, H) chains from two physically separable, antigenically distinct antibody populations, isolated from the same animal and having affinity for the same haptenic determinant, have been found to differ in their amino acid composition. Anti-Lac antibody light (B, L) chains have also been shown to be chemically heterogeneous, and contained populations of polypeptide chains possessing, and populations lacking methionine. The relevance of the observed structural heterogeneity of equine anti-Lac antibody to the problem of defining the mechanism of acquisition of immunological specificity is briefly discussed. PMID:4959973

Rockey, J H

1967-02-01

333

Antitubulin antibody in healthy adults and patients with infectious mononucleosis and its relationship to smooth muscle antibody (SMA).  

PubMed Central

Antibody to tubulin in man has been studied using a specific radioimmunoassay, affinity chromatography radioimmunoassay but markedly increased levels were noted in patients with infectious mononucleosis where the antibody was predominantly IgM in type. This finding was confirmed on fluorescence microscopy. Affinity chromatography purified antibody produced characteristic microtubular staining of fixed 3T3 cells, but in addition, produced weak staining of cryostat sections of rat tissue, similar in distribution to that of smooth muscle antibody. Our studies indicate that the IgM smooth muscle antibody found in infectious mononucleosis by IF techniques is at least in part due to an antitubulin antibody. Images FIG. 1 PMID:6993069

Mead, G M; Cowin, P; Whitehouse, J M

1980-01-01

334

Innovative monoclonal antibody therapies in multiple sclerosis  

Microsoft Academic Search

The recent years have witnessed great efforts in establishing new therapeutic options for multiple sclerosis (MS), especially for relapsing-remitting disease courses. In particular, the application of monoclonal antibodies provide innovative approaches allowing for blocking or depleting specific molecular targets, which are of interest in the pathogenesis of MS. While natalizumab received approval by the US Food and Drug Administration and

Ralf A. Linker; Bernd C. Kieseier

335

IgA Antibodies in Rett Syndrome  

ERIC Educational Resources Information Center

The level of IgA antibodies to gluten and gliadin proteins found in grains and to casein found in milk, as well as the level of IgG to gluten and gliadin, have been examined in 23 girls with Rett syndrome and 53 controls. Highly statistically significant increases were found for the Rett population compared to the controls. The reason for this…

Reichelt, K. L.; Skjeldal, O.

2006-01-01

336

IgA antibodies in Rett syndrome  

Microsoft Academic Search

The level of IgA antibodies to gluten and gliadin proteins found in grains and to casein found in milk, as well as the level of IgG to gluten and gliadin, have been examined in 23 girls with Rett syndrome and 53 controls. Highly statistically significant increases were found for the Rett population compared to the controls. The reason for this

K. L. Reichelt; O. Skjeldal

2006-01-01

337

Ebola Virus Antibodies in Fruit Bats, Bangladesh  

PubMed Central

To determine geographic range for Ebola virus, we tested 276 bats in Bangladesh. Five (3.5%) bats were positive for antibodies against Ebola Zaire and Reston viruses; no virus was detected by PCR. These bats might be a reservoir for Ebola or Ebola-like viruses, and extend the range of filoviruses to mainland Asia. PMID:23343532

Islam, Ariful; Yu, Meng; Anthony, Simon J.; Epstein, Jonathan H.; Khan, Shahneaz Ali; Khan, Salah Uddin; Crameri, Gary; Wang, Lin-Fa; Lipkin, W. Ian; Luby, Stephen P.; Daszak, Peter

2013-01-01

338

Antibody-Catalyzed Degradation of Cocaine  

Microsoft Academic Search

Immunization with a phosphonate monoester transition-state analog of cocaine provided monoclonal antibodies capable of catalyzing the hydrolysis of the cocaine benzoyl ester group. An assay for the degradation of radiolabeled cocaine identified active enzymes. Benzoyl esterolysis yields ecgonine methyl ester and benzoic acid, fragments devoid of cocaine's stimulant activity. Passive immunization with such an artificial enzyme could provide a treatment

Donald W. Landry; Kang Zhao; Ginger X.-Q. Yang; Michael Glickman; Taxiarchis M. Georgiadis

1993-01-01

339

Magic Bullets and Monoclonals: An Antibody Tale  

NSDL National Science Digital Library

FASEB Breakthroughs in Bioscience article. This most recent article describes the century of fundamental immunology research that led to todayĂÂs cutting edge monoclonal antibody therapies, used to treat millions of patients for several types of cancer, autoimmune and inflammatory disorders, and infectious disease.

Margie Patlak (Federation of American Societies for Experimental Biology Office of Public Affairs)

2010-07-12

340

Subcutaneous Administration of Monoclonal Antibodies in Oncology  

PubMed Central

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

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

2014-01-01

341

Antibodies to Squalene in Gulf War Syndrome  

Microsoft Academic Search

Gulf War Syndrome (GWS) is a multisystemic illness afflicting many Gulf War-era veterans. The molecular pathological basis for GWS has not been established. We sought to determine whether the presence of antibodies to squalene correlates with the presence of signs and symptoms of GWS. Participants in this blinded cohort study were individuals immunized for service in Desert Shield\\/Desert Storm during

Pamela B. Asa; Yan Cao; Robert F. Garry

2000-01-01

342

Developing recombinant antibodies for biomarker detection  

SciTech Connect

Monoclonal antibodies (mAbs) have an essential role in biomarker validation and diagnostic assays. A barrier to pursuing these applications is the reliance on immunization and hybridomas to produce mAbs, which is time-consuming and may not yield the desired mAb. We recommend a process flow for affinity reagent production that utilizes combinatorial protein display systems (eg, yeast surface display or phage display) rather than hybridomas. These systems link a selectable phenotype-binding conferred by an antibody fragment-with a means for recovering the encoding gene. Recombinant libraries obtained from immunizations can produce high-affinity antibodies (<10 nM) more quickly than other methods. Non-immune libraries provide an alternate route when immunizations are not possible, or when suitable mAbs are not recovered from an immune library. Directed molecular evolution (DME) is an integral part of optimizing mAbs obtained from combinatorial protein display, but can also be used on hybridoma-derived mAbs. Variants can easily be obtained and screened to increase the affinity of the parent mAb (affinity maturation). We discuss examples where DME has been used to tailor affinity reagents to specific applications. Combinatorial protein display also provides an accessible method for identifying antibody pairs, which are necessary for sandwich-type diagnostic assays.

Baird, Cheryl L.; Fischer, Christopher J.; Pefaur, Noah B.; Miller, Keith D.; Kagen, Jacob; Srivastava, Sudhir; Rodland, Karin D.

2010-10-01

343

Engineering aglycosylated antibody variants with immune effector functions  

E-print Network

Monoclonal antibodies have emerged as a promising class of therapeutics for the treatment of human disease, and in particular human cancer. While multiple mechanisms contribute to antibody efficacy, the engagement and ...

Sazinsky, Stephen L. (Stephen Lael)

2009-01-01

344

Neutralizing antibodies to HIV-1 induced by immunization  

PubMed Central

Most neutralizing antibodies act at the earliest steps of viral infection and block interaction of the virus with cellular receptors to prevent entry into host cells. The inability to induce neutralizing antibodies to HIV has been a major obstacle to HIV vaccine research since the early days of the epidemic. However, in the past three years, the definition of a neutralizing antibody against HIV has been revolutionized by the isolation of extremely broad and potent neutralizing antibodies from HIV-infected individuals. Considerable hurdles remain for inducing neutralizing antibodies to a protective level after immunization. Meanwhile, novel technologies to bypass the induction of antibodies are being explored to provide prophylactic antibody-based interventions. This review addresses the challenge of inducing HIV neutralizing antibodies upon immunization and considers notable recent advances in the field. A greater understanding of the successes and failures for inducing a neutralizing response upon immunization is required to accelerate the development of an effective HIV vaccine. PMID:23401570

McCoy, Laura E.

2013-01-01

345

Beyond natural antibodies: the power of in vitro display technologies  

PubMed Central

In vitro display technologies, best exemplified by phage and yeast display, were first described for the selection of antibodies some twenty years ago. Since that time a large number of antibodies, some with remarkable properties, have been selected and improved upon using these methods. The first antibodies derived using in vitro display methods are now in the clinic, with many more waiting in the wings. Here we discuss the scope of the technology, some of the powerful antibodies selected, and the future potential in a post-genomic world. Unique advantages offered by in vitro display technologies include the ability to carefully define selection conditions, allowing the derivation of antibodies recognizing predefined epitopes or conformations, the further improvement of selected antibodies, the potential for high throughput applications and the immediate availability of genes encoding the selected antibody. We anticipate that the high throughput potential of these technologies will soon lead to their use to select antibodies against all human proteins. PMID:21390033

Bradbury, Andrew R.M.; Sidhu, Sachdev; Dubel, Stefan; McCafferty, John

2011-01-01

346

Quantitative analysis of perivascular antibody distribution in solid tumors  

E-print Network

Monoclonal antibodies and proteins derived from them are an emerging class of anticancer therapeutics that have shown efficacy in a range of blood and solid tumors. Antibodies targeting solid tumors face considerable ...

Rhoden, John J. (John Joseph)

2013-01-01

347

Screening individual hybridomas by microengraving to discover monoclonal antibodies  

E-print Network

The demand for monoclonal antibodies (mAbs) in biomedical research is significant, but the current methodologies used to discover them are both lengthy and costly. Consequently, the diversity of antibodies available for ...

Ogunniyi, Adebola Oluwakayode

348

NCI Requests Cancer Targets for Monoclonal Antibody Production and Characterization  

Cancer.gov

In an effort to provide well-characterized monoclonal antibodies to the scientific community, NCI's Antibody Characterization Program requests cancer-related protein targets for affinity production and distribution.

349

NCI Requests Targets for Monoclonal Antibody Production and Characterization  

Cancer.gov

In an effort to provide well-characterized monoclonal antibodies to the scientific community, NCI's Antibody Characterization Program requests cancer-related protein targets for affinity production and distribution.

350

Monoclonal Antibodies against Cytosolic Thyroid Hormone Binding Protein.  

National Technical Information Service (NTIS)

The present invention related to the preparation of monoclonal antibodies having specific binding affinity for a cytoplasmic thyroid hormone binding protein. Heretofore, not even polyclonal antibodies to the cytosolic thyroid hormone binding protein exist...

S. Y. Cheng

1988-01-01

351

Sperm auto - antibodies and anti-nuclear antigen antibodies in chronic dioxin - exposed veterans.  

PubMed

The authors studied anti-nuclear antibodies (ANA) in 25 chronic dioxin - exposed veterans by IIF technics with Hep-2 cell line and sperm autoantibodies by agglutination test of Franklin-Dukes. The site of antibody binding on spermatozoon is detected by IIF test. The control group for ANA detection is 63 healthy persons of the same age as that of dioxin - exposed veterans and the control group for sperm autoantibodies is 36 healthy males of 28-63 years old, having 1-2 children. Obtained results show that the rate of ANA positive in veterans group is normal, and sperm auto-antibodies is also at normal range. The site of antibody binding on spermatozoon is predominantly head - head, rarely head - neck or tail - tail. PMID:8907229

Chinh, T T; Phi, P T; Thuy, N T

1996-02-01

352

CiteAb: a searchable antibody database that ranks antibodies by the number of times they have been cited  

PubMed Central

Background Research antibodies are used by thousands of scientists working in diverse disciplines, but it is common to hear concerns about antibody quality. This means that researchers need to carefully choose the antibodies they use to avoid wasting time and money. A well accepted way of selecting a research antibody is to identify one which has been used previously, where the associated data has been peer-reviewed and the results published. Description CiteAb is a searchable database which ranks antibodies by the number of times they have been cited. This allows researchers to easily find antibodies that have been used in peer-reviewed publications and the accompanying citations are listed, so users can check the data contained within the publications. This makes CiteAb a useful resource for identifying antibodies for experiments and also for finding information to demonstrate antibody validation. The database currently contains 1,400,000 antibodies which are from 90 suppliers, including 87 commercial companies and 3 academic resources. Associated with these antibodies are 140,000 publications which provide 306,000 antibody citations. In addition to searching, users can also browse through the antibodies and add their own publications to the CiteAb database. Conclusions CiteAb provides a new way for researchers to find research antibodies that have been used successfully in peer-reviewed publications. It aims to assist these researchers and will hopefully help promote progress in many areas of life science research. PMID:24528853

2014-01-01

353

Antibodies and neuronal autoimmune disorders of the CNS.  

PubMed

We review the neuronal antibodies described in CNS disorders in order to clarify their diagnostic value, emphasize potentials pitfalls and limitations in the diagnosis of paraneoplastic neurological syndromes (PNS), and examine the current evidence for a possible pathogenic role. We propose to classify the neuronal antibodies associated with syndromes resulting from CNS neuronal dysfunction into two groups according to the location of the antigen: inside the neuron or in the cell membrane. Group I includes antibodies which target intracellular antigens and probably are not pathogenic. They are further subdivided into three groups. Group Ia comprises well-characterized onconeural antibodies (Hu (ANNA1), Yo (PCA1), Ri (ANNA2), CV2 (CRMP5), amphiphysin, Ma2) that are useful for the diagnosis of PNS. Group Ib antibodies (SOX and ZIC) are cancer-specific but there is no evidence that the immune response is in any way pathogenically related to the PNS. Antibodies in group Ic (glutamic acid decarboxylase (GAD), adenylate kinase 5 and Homer 3) identify non-PNS: stiff-person syndrome (SPS), cerebellar ataxia, and limbic encephalitis (LE). Group II antibodies recognize neuronal surface antigens. Antibodies in group IIa associate with characteristic CNS syndromes but their detection does not indicate that the disorder is paraneoplastic. Antibodies to potassium channels, AMPA and GABA(B) receptors are associated with LE, NMDA receptor antibodies identify a well-defined encephalitis, and antibodies against glycine receptors associate with SPS with encephalitis. A pathogenic role of the antibodies is suggested by the response of symptoms to immunotherapy and the correlation between antibody titers and neurological outcome. Lastly, Group IIb includes antibodies that are found in patients with paraneoplastic cerebellar ataxia associated with lung cancer (P/Q type calcium channels antibodies) or Hodgkin disease (metabotropic glutamate receptor type 1 antibodies). PMID:20035430

Graus, Francesc; Saiz, Albert; Dalmau, Josep

2010-04-01

354

FLT3 Antibody-Based Therapeutics for Leukemia Therapy  

Microsoft Academic Search

Antibodies represent a unique class of therapeutics because of their high specificity toward a defined target antigen. Recent\\u000a clinical success with antibody-based cancer therapeutics has led to an upsurge in the development of these agents. Antibodies\\u000a directed against FLT3 represent a promising approach for the treatment of human leukemia. We discuss some basic aspects of\\u000a antibody-based cancer therapeutics, including their

Yiwen Li; Zhenping Zhu

2005-01-01

355

Lattice Formation in Complement Fixation: Studies with Univalent Rabbit Antibody  

Microsoft Academic Search

Hybrid univalent 6.5S antibody molecules, formed by recombination of half-molecules of rabbit antibody to ovalbumin with those of normal rabbit gamma G-globulin, fail to fix complement in reactions with homologous antigen. Such hybrid molecules, however, block complement fixation by intact antibody to ovalbumin. Molecules of antibody reconstituted in the absence of other protein retain the capacity to fix complement. The

H. H. Fudenberg; R. Hong; A. Nisonoff

1965-01-01

356

Development of murine monoclonal antibodies to methamphetamine and methamphetamine analogues  

Microsoft Academic Search

Methamphetamine and ecstasy are addictive drugs that cause major health problems in young people. Here we report on the development of high-affinity monoclonal antibodies to methamphetamine and its analogues, which may constitute powerful tools for antibody-based therapy. Six haptens, methamphetamine and ecstasy analogues, were synthesized, linked to a carrier protein and injected into mice. Several specific monoclonal antibodies were subsequently

Yannic Danger; Caroline Gadjou; Anne Devys; Hervé Galons; Dominique Blanchard; Gilles Folléa

2006-01-01

357

Precipitating Antibodies in the Carpet Snake against Parasitic Nematodes  

Microsoft Academic Search

To our knowledge very little attention has been paid to the formation or presence of precipitating antibodies in the cold-blooded vertebrates. Indications of active antibody formation have nevertheless been reported in frogs, turtles and fish; however, these estimations were made using either agglutination or anaphylaxis1. This communication is a survey of naturally occurring antibodies in the carpet snake (Morelia spilotes

H. Timourian; C. Dobson; J. F. A. Sprent

1961-01-01

358

Breaking the one antibody-one target axiom  

Microsoft Academic Search

Studies at the interface of chemistry and biology have allowed us to develop an immunotherapeutic approach called chemically programmed antibodies (cpAbs), which combines the merits of traditional small-molecule drug design with immunotherapy. In this approach, a catalytic antibody catalyzes the covalent conjugation of a small molecule or peptide to the active site of the antibody, effectively recruiting the binding specificity

Fang Guo; Sanjib Das; Barbara M. Mueller; Carlos F. Barbas III; Richard A. Lerner; Subhash C. Sinha

2006-01-01

359

Monoclonal antibodies and method for detecting dioxins and dibenzofurans  

DOEpatents

Compositions of matter are described which include five monoclonal antibodies that react with dioxins and dibenzofurans, and the five hybridomas that produce these monoclonal antibodies. In addition, a method for the use of these antibodies in a sensitive immunoassay for dioxins and dibenzofurans is given, which permits detection of these pollutants in samples at concentrations in the range of a few parts per billion.

Vanderlaan, Martin (San Ramon, CA); Stanker, Larry H. (Livermore, CA); Watkins, Bruce E. (Livermore, CA); Bailey, Nina R. (Berkley, CA)

1989-01-01

360

Genetically engineered antibodies and their application to brain delivery  

Microsoft Academic Search

Techniques of genetic engineering and expression have been applied to the production of antibodies in a variety of expression systems. Combinatorial libraries produced in bacteriophage may present an alternative to animal immunization as a source of antigen-binding specificities. Transfectomas which express genetically engineered antibody genes provide one approach to overcoming some of the limitations inherent in classical monoclonal antibodies. Novel

Sherie L. Morrison; Seung-Uon Shin

1995-01-01

361

Monoclonal antibody specific for a pigmentation associated antigen  

SciTech Connect

Monoclonal antibody TA99, which specifically binds to a pigmentation associated antigen present on melanoma cells is described. Additionally, the hybridoma cell line deposited with the ATCC under Accession Number HB 8704 from which the antibody is derived, as well as methods for using the antibody are described.

Thomson, T.M.; Mattes, M.J.; Old, L.J.; Lloyd, K.O

1989-01-17

362

Detection of Antibodies to Melanocytes in Vitiligo by Specific Immunoprecipitation  

Microsoft Academic Search

Immunoprecipitation was used to assay for antibodies to normal human melanocytes in the sera of 12 patients with common vitiligo and 12 normal individuals. The procedure is based on the specific immunoprecipitation using protein A-sepharose of antibodies binding to detergent-soluble, radioiodinated macromolecules of normal human melanocytes grown in culture. Antibodies to melanocytes were found in all 12 patients with vitiligo

Gail K. Naughton; Magdalena Eisinger; Jean-Claude Bystryn

1983-01-01

363

Continuous cultures of fused cells secreting antibody of predefined specificity  

Microsoft Academic Search

THE manufacture of predefined specific antibodies by means of permanent tissue culture cell lines is of general interest. There are at present a considerable number of permanent cultures of myeloma cells1,2 and screening procedures have been used to reveal antibody activity in some of them. This, however, is not a satisfactory source of monoclonal antibodies of predefined specificity. We describe

G. Köhler; C. Milstein

1975-01-01

364

Antibody drug conjugates — Trojan horses in the war on cancer  

Microsoft Academic Search

Antibody drug conjugates (ADCs) consist of an antibody attached to a cytotoxic drug by means of a linker. ADCs provide a way to couple the specificity of a monoclonal antibody (mAb) to the cytotoxicity of a small-molecule drug and, therefore, are promising new therapies for cancer. ADCs are prodrugs that are inactive in circulation but exert their cytotoxicity upon binding

U. Iyer; V. J. Kadambi

365

Bispecific Antibodies: Molecules That Enable Novel Therapeutic Strategies  

Microsoft Academic Search

Bispecific antibodies are unique in the sense that they can bind simultaneously two different antigens. This property enables the development of therapeutic strategies that are not possible with conventional monoclonal antibodies. The large panel of imaginative bispecific antibody formats that has been developed reflects the strong interest for these molecules. Although in many cases the manufacturing of clinical grade material

Nicolas Fischer; Olivier Léger

2007-01-01

366

Lymphocyte-reactive antibodies in acquired immune deficiency syndrome  

Microsoft Academic Search

Antilymphocyte antibodies were studied using the Terasaki microcytotoxicity technique in 21 gay patients including 7 with Kaposi's sarcoma, 5 with opportunistic infection, and 9 with lymphadenopathy syndrome. A significant increase in lymphocyte-reactive antibody was noted in 61% of this group. Similar studies using serum from 25 apparently healthy gay males showed lymphocytotoxic antibody in only one instance. When isolated T-cell

Ralph C. Williams; Henry Masur; Thomas J. Spira

1984-01-01

367

Single-chain antibodies as diagnostic tools and therapeutic agents.  

PubMed

Over three decades after the generation of the first mouse monoclonal antibodies by Kohler and Milstein, recombinant antibodies are the fastest growing class of therapeutic proteins. Furthermore, antibodies are key detection reagents in research and diagnostics. Technology improvements have provided several approaches to manufacturing human antibodies with high affinity for biologically relevant targets. Approximately 300 development programs for therapeutic antibodies have been reported in industrial and academic laboratories, and this clearly demonstrates the expectations towards antibody technology. Antibody fragments are a subclass with growing clinical importance. This review focuses on single-chain antibodies as one of the smallest possible format for recombinant antibodies and their use as diagnostic tools and therapeutic agents. We describe the structure, selection and production of single-chain antibodies. Furthermore, we review current applications of antibody fragments focusing on thrombus targeting using fibrin- and platelet-specific single-chain antibodies as well as describing novel noninvasive imaging approaches for the diagnosis of thrombosis and inflammation. PMID:19492141

Hagemeyer, Christoph E; von Zur Muhlen, Constantin; von Elverfeldt, Dominik; Peter, Karlheinz

2009-06-01

368

Behavioral and Psychological Responses to HIV Antibody Testing.  

ERIC Educational Resources Information Center

Considers effects of informing individuals of their antibody status as determined by human immunodeficiency virus (HIV) antibody testing. Reviews research examining changes in psychological distress and in behaviors associated with HIV infections among individuals who have undergone antibody testing. Identifies methodological issues in studying…

Jacobsen, Paul B.; And Others

1990-01-01

369

A Simple Model System to Demonstrate Antibody Structure and Functions.  

ERIC Educational Resources Information Center

A model that can be used to show the arrangement of light and heavy chains, disulfide linkages, domains, and subclass variations in antibodies is given. It can be constructed and modified to illustrate Fab, F(ab')2, and Fc fragments, single domain and bifunctional antibodies, and labeling of antibodies. (Author)

O'Kennedy, Richard

1991-01-01

370

Anti-DNA antibodies in the primary antiphospholipid syndrome (PAPS)  

PubMed

Primary antiphospholipid syndrome (PAPS) is considered a distinct entity from SLE and patients with PAPS are generally regarded as being dsDNA antibody negative. Levels of IgG and IgM ss and ds DNA antibodies were measured by ELISA in 30 patients who fulfilled the criteria for the diagnosis of PAPS. We compared these patients with 20 normal controls and seven patients with idiopathic SLE. We also examined all the sera for anti-nuclear antibodies by Hep-2 cells and for dsDNA antibodies by Crithidia. We found that 16 patients with PAPS had antibodies to ss and/or dsDNA. Only three of the 16 positive patients had both IgG and IgM anti-DNA antibodies. Twelve patients had anti-nuclear antibodies, but only two were weakly positive for dsDNA antibodies by Crithidia immunofluorescence. Eleven out of 30 patients with PAPS had IgM anti-dsDNA antibodies compared to two out of the seven SLE patients. The PAPS patients with anti-DNA antibodies were clinically indistinguishable from the PAPS patients without antibodies against DNA. Our results show that 53% of patients with PAPS had antibodies to DNA which supports the view that PAPS and SLE are probably overlapping disorders. PMID:8495254

Ehrenstein, M R; Swana, M; Keeling, D; Asherson, R; Hughes, G R; Isenberg, D A

1993-05-01

371

FINAL REPORT. ENGINEERED ANTIBODIES FOR MONITORING OF POLYNUCLEAR AROMATIC HYDROCARBONS  

EPA Science Inventory

This project was conducted to remove the major barrier to the timely development and use of more versatile antibody-based detection and sample cleanup methods. The main objective was to adapt combinatorial antibody library and antibody engineering methods for preparing a panel of...

372

Have we overestimated the benefit of human(ized) antibodies?  

PubMed Central

The infusion of animal-derived antibodies has been known for some time to trigger the generation of antibodies directed at the foreign protein as well as adverse events including cytokine release syndrome. These immunological phenomena drove the development of humanized and fully human monoclonal antibodies. The ability to generate human(ized) antibodies has been both a blessing and a curse. While incremental gains in the clinical efficacy and safety for some agents have been realized, a positive effect has not been observed for all human(ized) antibodies. Many human(ized) antibodies trigger the development of anti-drug antibody responses and infusion reactions. The current belief that antibodies need to be human(ized) to have enhanced therapeutic utility may slow the development of novel animal-derived monoclonal antibody therapeutics for use in clinical indications. In the case of murine antibodies, greater than 20% induce tolerable/negligible immunogenicity, suggesting that in these cases humanization may not offer significant gains in therapeutic utility. Furthermore, humanization of some murine antibodies may reduce their clinical effectiveness. The available data suggest that the utility of human(ized) antibodies needs to be evaluated on a case-by-case basis, taking a cost-benefit approach, taking both biochemical characteristics and the targeted therapeutic indication into account. PMID:20935511

Getts, Meghann T; McCarthy, Derrick P; Chastain, Emily ML; Miller, Stephen D

2010-01-01

373

Envelope Deglycosylation Enhances Antigenicity of HIV-1 gp41 Epitopes for Both Broad Neutralizing Antibodies and Their Unmutated Ancestor Antibodies  

E-print Network

The HIV-1 gp41 envelope (Env) membrane proximal external region (MPER) is an important vaccine target that in rare subjects can elicit neutralizing antibodies. One mechanism proposed for rarity of MPER neutralizing antibody ...

Ma, Ben-Jiang; Alam, S. Munir; Go, Eden P.; Lu, Xiaozhi; Desaire, Heather; Tomaras, Georgia D.; Bowman, Cindy; Sutherland, Laura L.; Scearce, Richard M.; Santra, Sampa; Letivn, Norman L.; Kepler, Thomas B.; Liao, Hua-Xin; Haynes, Barton F.

2011-09-01

374

Galactosylation variations in marketed therapeutic antibodies.  

PubMed

There are currently ~25 recombinant full-length IgGs (rIgGs) in the market that have been approved by regulatory agencies as biotherapeutics to treat various human diseases. Most of these are based on IgG1k framework and are either chimeric, humanized or human antibodies manufactured using either Chinese hamster ovary (CHO) cells or mouse myeloma cells as the expression system. Because CHO and mouse myeloma cells are mammalian cells, rIgGs produced in these cell lines are typically N-glycosylated at the conserved asparagine (Asn) residues in the CH2 domain of the Fc, which is also the case with serum IgGs. The Fc glycans present in these rIgGs are for the most part complex biantennary oligosaccharides with heterogeneity associated with the presence or the absence of several different terminal sugars. The major Fc glycans of rIgGs contain 0 or 1 or 2 (G0, G1 and G2, respectively) terminal galactose residues as non-reducing termini and their relative proportions may vary depending on the cell culture conditions in which they were expressed. Since glycosylation is strongly associated with antibody effector functions and terminal galactosylation may affect some of those functions, a panel of commercially available therapeutic rIgGs expressed in CHO cells and mouse myeloma cells were examined for their galactosylation patterns. The results suggest that the rIgGs expressed in CHO cells are generally less galactosylated compared to the rIgGs expressed in mouse myeloma cells. Accordingly, rIgGs produced in CHO cells tend to contain higher levels of G0 glycans compared with rIgGs produced in mouse myeloma cell lines. Despite the apparent wide variability in galactose content, adverse events or safety issues have not been associated with specific galactosylation patterns of therapeutic antibodies. Nevertheless, galactosylation may have an effect on the mechanisms of action of some therapeutic antibodies (e.g., effector pathways) and hence further studies to assess effects on product efficacy may be warranted for such antibodies. For antibodies that do not require effector functions for biological activity, however, setting a narrow specification range for galactose content may be unnecessary. PMID:22531450

Raju, T Shantha; Jordan, Robert E

2012-01-01

375

New choices for patients needing kidney transplantation across antibody barriers.  

PubMed

Antibodies in the blood of a kidney transplant recipient can provide a barrier to transplantation, which is additional to the usual possibility of cellular rejection. The antibodies most frequently encountered are ABO (blood group) and human leucocyte antigen (HLA) (tissue-type) antibodies. About 250 living donor transplants each year in the United Kingdom have been stopped because of an antibody barrier. It is now possible to offer a choice of treatment modalities to these people, including exchange transplantation and antibody-incompatible transplantation. It is likely that both schemes will complement each other and both are available in the United Kingdom. PMID:18498573

Higgins, Rob; Hathaway, Mark; Lowe, David; Zehnder, Daniel; Krishnan, Nithya; Hamer, Rizwan; Briggs, David

2008-06-01

376

Production of antibodies which recognize opiate receptors on murine leukocytes  

SciTech Connect

An antibody has been developed which recognizes opiate receptors on cells of the immune system. This antibody blocks specific binding of the radiolabeled opiate receptor ligand, /sup 3/H-dihydromorphine, to receptors on murine splenocytes. Additionally, the anti-receptor antibody competes with ..beta..-endorphin, meta-enkephalin, and naloxone for the same binding site on the leukocytes. Moreover, the anti-receptor antibody possesses agonist activity similar to ..beta..-endorphin in suppressing cAMP production by lymphocytes. These results suggest the development of an antibody which recognizes classical opiate receptors on cells of the immune system.

Carr, D.J.J.; Bost, K.L.; Blalock, J.E.

1988-01-01

377

Antibodies to idiotypes of isologous immunoglobulins  

PubMed Central

To determine if the immunoglobulins (Igs) capable of eliciting the formation of isologous anti-idiotypic antibodies are rare exceptions, BABL/c mice were immunized with five myeloma proteins of BALB/c origin. Anti-idiotypes were produced against all but one. The idiotype of the exception (T15) is remarkably abundant in BALB/c mice, whose unresponsiveness is probably due to tolerance. Nevertheless, prolonged immunization with T15 finally induced the formation of isologous antibodies that seemed largely to be specific for IgA proteins, especially those with k-light-chains; the reactions of a few of these isologous antisera with T15 were slightly inhibited by phosphorylcholine, suggesting that some anti-idiotypes were probably formed even to this unusually prevalent idiotype. It is likelythat under appropriate conditions almost any myeloma protein can elicit isologous anti-idiotypes. PMID:805211

1975-01-01

378

Antibody production in Syphacia obvelata infected mice.  

PubMed

Antibody response to Syphacia obvelata infection was observed in AKR/J mice by ELISA. Experimental infection with the pinworm eggs showed the presence of specific IgG against S. obvelata somatic antigens at 12 days postinfection, and that it increased steadily thereafter. Sera of S. obvelata-infected mice showed cross-reactivity with somatic antigens of other Syphacia species such as S. mesocriceti and S. muris, but not with Aspiculuris asiatica. Western blotting of S. obvelata antigen with sera of S. obvelata-infected mice showed a corresponding increase in the number of bands during the course of infection. Infected mice showed significantly higher antibody production to sheep red blood cells than the uninfected control mice. Thus, S. obvelata infection is shown to alter the humoral response to nonparasitic antigenic stimuli. These observations indicate that infection by helminths, which apparently do not produce clinical symptoms, might modulate the immune system of the host and, therefore, affect experimental results. PMID:7623197

Sato, Y; Ooi, H K; Nonaka, N; Oku, Y; Kamiya, M

1995-08-01

379

Primary antibody deficiency and Crohn's disease  

PubMed Central

Five patients with primary antibody deficiency were investigated because of intermittent but persistent diarrhoea of several years duration despite immunoglobulin replacement therapy. We found no evidence of Giardia lambia or other intestinal pathogens to explain their gastrointestinal symptoms. All five had definite radiological evidence of small bowel Crohn's disease and three had histological specimens available with abnormalities consistent with Crohn's disease. One patient had a non-caseating granuloma in an oral ulcer. A second patient with stricturing disease in the small bowel had a mucosal inflammatory infiltrate with non-caseating granulomas. A third had transmural inflammation but no granulomas. All five patents were diagnosed as having Crohn's disease and have responded symptomatically to steroid therapy.???Keywords: antibody deficiency; Crohn's disease PMID:10448496

Conlong, P; Rees, W.; Shaffer, J; Nicholson, D.; Jewell, D.; Heaney, M.; Jones, A.; Snowden, N.

1999-01-01

380

Antibody-mediated cofactor-driven reactions  

DOEpatents

Chemical reactions capable of being rate-enhanced by auxiliary species which interact with the reactants but do not become chemically bound to them in the formation of the final product are performed in the presence of antibodies which promote the reactions. The antibodies contain regions within their antigen binding sites which recognize the auxiliary species in a conformation which promotes the reaction. The antigen binding site frequently recognizes a particular transition state complex or other high energy complex along the reaction coordinate, thereby promoting the progress of the reaction along the desired route as opposed to other less favorable routes. Various classes of reaction together with appropriate antigen binding site specificities tailored for each are disclosed.

Schultz, Peter G. (Oakland, CA)

1993-01-01

381

Detection of antinuclear antibodies in SLE.  

PubMed

The antinuclear antibodies (ANA) also known as antinuclear factors (ANF) are unwanted molecules which bind and destroy certain structures within the nucleus. In systemic lupus erythematosus (SLE), they are produced in excess; hence their detection in the blood of patients is important for diagnosis and monitoring of the disease. Several methods are available which can be used to detect ANA; nevertheless, indirect immunofluorescence antinuclear antibody test (IF-ANA) is considered a "reference method" for their detection. Though IF-ANA is relatively easier to perform, its interpretation requires considerable skill and experience. The chapter therefore is aimed to provide comprehensive details to readers, not only about its methodology but also the result interpretation and reporting aspects of IF-ANA. PMID:24497352

Kumar, Yashwant; Bhatia, Alka

2014-01-01

382

Innovative Monoclonal Antibody Therapies in Multiple Sclerosis  

PubMed Central

The recent years have witnessed great efforts in establishing new therapeutic options for multiple sclerosis (MS), especially for relapsing–remitting disease courses. In particular, the application of monoclonal antibodies provide innovative approaches allowing for blocking or depleting specific molecular targets, which are of interest in the pathogenesis of MS. While natalizumab received approval by the US Food and Drug Administration and the European Medicines Agency in 2006 as the first monoclonal antibody in MS therapy, rituximab, alemtuzumab, and daclizumab were successfully tested for relapsing-remitting MS in small cohorts in the meantime. Here, we review the data available from these recent phase II trials and at the same time critically discuss possible pitfalls which may be relevant for clinical practice. The results of these studies may not only broaden our therapeutic options in the near future, but also provide new insights into disease pathogenesis. PMID:21180564

Kieseier, Bernd C.

2008-01-01

383

Next generation and biosimilar monoclonal antibodies  

PubMed Central

The Next Generation and Biosimilar Monoclonal Antibodies: Essential Considerations Towards Regulatory Acceptance in Europe workshop, organized by the European Centre of Regulatory Affairs Freiburg (EUCRAF), was held February 3–4, 2011 in Freiburg, Germany. The workshop attracted over 100 attendees from 15 countries, including regulators from 11 agencies, who interacted over the course of two days. The speakers presented their authoritative views on monoclonal antibodies (mAbs) as attractive targets for development, the experience to date with the regulatory process for biosimilar medicinal products, the European Medicines Agency draft guideline on biosimilar mAbs, as well as key elements in the development of mAbs. Participants engaged in many lively discussions, and much speculation on the nature of the quality, non-clinical and clinical requirements for authorization of biosimilar mAbs. PMID:21487235

2011-01-01

384

Age-dependent variations of antibody avidity.  

PubMed Central

Age-dependent variations of antibody avidity were studied in the C3HeB/FeJ mouse. Spleen cells from donors of different ages (10--720 days) were transferred and stimulated with TNP-HRBC in lethally irradiated syngenic recipients. The anti-TNP antibody response of the donor cells was estimated from the number of direct PFC per recipient spleen by the Jerne technique with TNP-SRBC. Avidity of the antibodies secreted by PFC was evaluated from the amount of added TNP-BSA that inhibited 50% of the anti-TNP PFC. Under these experimental conditions allowing the exclusion of any influence of the donor milieu during the immune response, age-dependent variations of the antibody response and avidity could be attributed to changes in the donor spleen cell population. Avidity was found to increase with the response and to vary parabolically with age. After appropriate correction of the number of PFC to make it independent from age, avidity values were fitted by a multiple curvilinear regression in which the independent variables playing a significant role were the corrected number of PFC in its linear term and the age in its linear and quadratic terms. From comparison of the standard coefficients of this regression, the observed variations of avidity could be attributed in part (82%) to the response and in part (18%) to the age. For any value of response, avidity increased 15-fold from day 10 to reach a maximum at day 110 and then declined 5-fold at the age of 720 days. Heterogeneity of avidity also changed parabolically with age as high avidity classes were present in adulthood and absent at 10 and 720 days. PMID:361545

Doria, G; D'Agostaro, G; Poretti, A

1978-01-01

385

[Predictive biomarkers for anti-EGFR antibodies].  

PubMed

The clinical significance of KRAS gene testing prior to using anti-epidermal growth factor receptor(EGFR)antibodies for colorectal cancer patients has been established in past randomized clinical trials. Thus, testing for the 7 most common mutations of KRAS codons 12 and 13 is now recommended as a clinical practice. However, pooled analysis of randomized controlled studies in Western countries in patients treated with cetuximab has suggested that patients with tumors showing the KRAS p. G13D mutation[a glycine(G)to aspartate(D)transition mutation] have longer overall survival and progression-free survival when compared to patients with other KRAS mutations. Furthermore, even among patients whose tumors are wild-type for KRAS codons 12 and 13, response rates are only 13~17% for anti-EGFR antibody monotherapy. These facts suggest that additional activating mutations in the RAS-RAF-MAPK or PI3K-AKT-mTOR pathways may also confer resistance to anti-EGFR antibody therapies. Indeed, recent retrospective studies have shown that mutations in KRAS codon 61 and 146, BRAF, NRAS, and PIK3CA may also predict resistance to anti-EGFR antibodies in colorectal cancer patients. On the other hand, the continuous use of anti-EGFR therapies for KRAS wild-type patients may lead to secondary resistance. Acquired EGFR or KRAS mutations have occasionally been detected among specimens from these patients. We review the latest personalized therapy available for colorectal cancer patients using KRAS mutational testing. We also illustrate future perspectives for patient selection using KRAS, BRAF, NRAS, PIK3CA, and other mutations. PMID:23152013

Bando, Hideaki; Yoshino, Takayuki

2012-11-01

386

Antineutrophil Cytoplasmic Antibodies, Autoimmune Neutropenia, and Vasculitis  

PubMed Central

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

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

2011-01-01

387

The biotechnology and applications of antibody engineering  

Microsoft Academic Search

The exquisite specificity of monoclonal antibodies (MAb) has long provided the potential for creating new reagents for the\\u000a in vivo delivery of therapeutic drugs or toxins to defined cellular target sites or improved methods of diagnosis. However,\\u000a many difficulties associated with their production, affinity, specificity, and use in vivo have largely confined their application\\u000a to research or in vitro diagnostics.

Ralph Rapley

1995-01-01

388

Stiff-person syndrome with amphiphysin antibodies  

PubMed Central

Background: Stiff-person syndrome (SPS), formerly Stiff-man syndrome, is a rare autoimmune disease usually exhibiting severe spasms and thoracolumbar stiffness, with very elevated glutamic acid decarboxylase antibodies (GAD Ab). A paraneoplastic variant, less well characterized, is associated with amphiphysin antibodies (amphiphysin Ab). The objective of this study was to identify distinctive clinical features of amphiphysin Ab-associated SPS. Methods: Records associated with 845 sera tested in the Yale SPS project were examined, and 621 patients with clinically suspected SPS were included in the study. Clinical characteristics were assessed with correction for multiple comparisons. Results: In all, 116 patients had GAD antibodies and 11 patients had amphiphysin Ab; some clinical information was available for 112 and 11 of these patients, respectively. Patients with amphiphysin Ab-associated SPS were exclusively female; mean age was 60. All except one had breast cancer; none had diabetes. Compared to patients with GAD Ab-associated SPS, those with amphiphysin Ab were older (p = 0.02) and showed a dramatically different stiffness pattern (p < 0.0000001) with cervical involvement more likely, p ? 0.001. Electromyography showed continuous motor unit activity or was reported positive in eight. Benzodiazepines at high dose (average 50 mg/day diazepam) were partially effective. Four patients were steroid responsive and tumor excision with chemotherapy produced marked clinical improvement in three of five patients. Conclusions: Amphiphysin Ab-associated stiff-person syndrome is strongly associated with cervical region stiffness, female sex, breast cancer, advanced age, EMG abnormalities, and benzodiazepine responsiveness. The condition may respond to steroids and can dramatically improve with cancer treatment. GLOSSARY EAE = experimental autoimmune encephalitis; GAD Ab = glutamic acid decarboxylase antibodies; ICC = immunocytochemistry; PERM = progressive variant with encephalomyelitis, rigidity, and myoclonus; SPS = stiff-person syndrome. PMID:18971449

Murinson, Beth B.; Guarnaccia, Joseph B.

2008-01-01

389

Antibody-Based Vascular Tumor Targeting  

Microsoft Academic Search

\\u000a The inhibition of angiogenesis represents a major step toward a more selective and better-tolerated therapy of cancer. An\\u000a alternative way to take advantage of a tumor’s absolute dependence on a functional neovasculature is illustrated by the strategy\\u000a of “antibody-based vascular tumor targeting.” This technology aims at the selective delivery of bioactive molecules to the\\u000a tumor site by their conjugation to

Christoph Schliemann; Dario Neri

390

Antiphospholipid antibodies and Mycoplasma pneumoniae infection.  

PubMed Central

Anticardiolipin antibody levels were measured in 57 patients with Mycoplasma pneumoniae infection and 21 patients with other infections. Significantly more patients in the mycoplasma group had increased IgM and IgG anticardiolipin. Within the mycoplasma group significantly higher titres were found in patients with severe infection (assessed by need for hospital admission) and in patients with cold agglutinins. A tendency for particularly high titres to occur in patients with extra-pulmonary complications was identified. PMID:2371184

Snowden, N.; Wilson, P. B.; Longson, M.; Pumphrey, R. S.

1990-01-01

391

Modulating the pharmacokinetics of therapeutic antibodies  

Microsoft Academic Search

With the advent of antibody fragments and alternative binding scaffolds, that are devoid of Fc-regions, strategies to increase\\u000a the half-life of small proteins are becoming increasingly important. Currently, the established method is chemical PEGylation,\\u000a but more elaborate approaches are being described such as polysialylation, amino acid polymers and albumin-binding derivatives.\\u000a This article reviews the main strategies for pharmacokinetic enhancement, primarily

A. Constantinou; C. Chen; M. P. Deonarain

2010-01-01

392

Monoclonal antibody targets, kills leukemia cells  

Cancer.gov

Researchers at the University of California, San Diego Moores Cancer Center have identified a humanized monoclonal antibody that targets and directly kills chronic lymphocytic leukemia (CLL) cells. The findings, published in the online Early Edition of the Proceedings of the National Academy of Sciences on March 25, 2013 represent a potential new therapy for treating at least some patients with CLL, the most common type of blood cancer in the United States.

393

Studies on production of anticollagen antibodies in silicosis  

SciTech Connect

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.

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

1993-01-01

394

Discovery of internalizing antibodies to tumor antigens from phage libraries  

PubMed Central

Phage antibody technology can be used to generate human antibodies to essentially any antigen. Many therapeutic target antigens are cell surface receptors, which can be challenging targets for antibody generation. In addition, for many therapeutic applications, one needs antibodies that not only bind the cell surface receptor but that also are internalized into the cell upon binding. This allows use of the antibody to deliver a range of payloads into the cell to achieve a therapeutic effect. In this chapter we describe how human phage antibody libraries can be selected directly on tumor cell lines to generate antibodies that bind cell surface receptors and which upon binding are rapidly internalized into the cell. Specific protocols show how to: 1) directly select cell binding and internalizing antibodies from human phage antibody libraries; 2) screen the phage antibodies in a high throughput flow cytometry assay for binding to the tumor cell line used for selection; 3) identify the antigen bound by the phage antibody using immunoprecipitation and mass spectrometry; and 4) direct cell binding and internalizing selections to a specific tumor antigen by sequential selection on a tumor cell line followed by selection on yeast displaying the target tumor antigen on the yeast surface. PMID:22208981

Zhou, Yu; Marks, James D

2014-01-01

395

IBC's 23rd Antibody Engineering and 10th Antibody Therapeutics Conferences and the Annual Meeting of The Antibody Society: December 2-6, 2012, San Diego, CA.  

PubMed

Now in its 23rd and 10th years, respectively, the Antibody Engineering and Antibody Therapeutics conferences are the Annual Meeting of The Antibody Society. The scientific program covers the full spectrum of challenges in antibody research and development from basic science through clinical development. In this preview of the conferences, the chairs provide their thoughts on sessions that will allow participants to track emerging trends in (1) the development of next-generation immunomodulatory antibodies; (2) the complexity of the environment in which antibodies must function; (3) antibody-targeted central nervous system (CNS) therapies that cross the blood brain barrier; (4) the extension of antibody half-life for improved efficacy and pharmacokinetics (PK)/pharmacodynamics (PD); and (5) the application of next generation DNA sequencing to accelerate antibody research. A pre-conference workshop on Sunday, December 2, 2012 will update participants on recent intellectual property (IP) law changes that affect antibody research, including biosimilar legislation, the America Invents Act and recent court cases. Keynote presentations will be given by Andreas Plückthun (University of Zürich), who will speak on engineering receptor ligands with powerful cellular responses; Gregory Friberg (Amgen Inc.), who will provide clinical updates of bispecific antibodies; James D. Marks (University of California, San Francisco), who will discuss a systems approach to generating tumor targeting antibodies; Dario Neri (Swiss Federal Institute of Technology Zürich), who will speak about delivering immune modulators at the sites of disease; William M. Pardridge (University of California, Los Angeles), who will discuss delivery across the blood-brain barrier; and Peter Senter (Seattle Genetics, Inc.), who will present his vision for the future of antibody-drug conjugates. For more information on these meetings or to register to attend, please visit www.IBCLifeSciences.com/AntibodyEng or call 800-390-4078. Members of The Antibody Society and mAbs journal subscribers receive a 20% discount for meeting registration. To obtain this discount, email kdostie@ibcusa.com. mAbs is the official therapeutics journal of The Antibody Society and offers a discounted subscription to Society members for $49. PMID:23007482

Marquardt, John; Begent, Richard H J; Chester, Kerry; Huston, James S; Bradbury, Andrew; Scott, Jamie K; Thorpe, Philip E; Veldman, Trudi; Reichert, Janice M; Weiner, Louis M

2012-01-01

396

Polyclonal antibody to soman-tyrosine  

PubMed Central

Soman forms a stable, covalent bond with tyrosine 411 of human albumin, with tyrosines 257 and 593 in human transferrin, and with tyrosine in many other proteins. The pinacolyl group of soman is retained, suggesting that pinacolyl methylphosphonate bound to tyrosine could generate specific antibodies. Tyrosine in the pentapeptide RYGRK was covalently modified with soman simply by adding soman to the peptide. The phosphonylated-peptide was linked to keyhole limpet hemocyanin, and the conjugate was injected into rabbits. The polyclonal antiserum recognized soman-labeled human albumin, soman-mouse albumin, and soman human transferrin, but not non-phosphonylated control proteins. The soman-labeled tyrosines in these proteins are surrounded by different amino acid sequences, suggesting that the polyclonal recognizes soman-tyrosine independent of the amino acid sequence. Antiserum obtained after 4 antigen injections over a period of 18 weeks was tested in a competition ELISA where it had an IC50 of 10?11 M. The limit of detection on Western blots was 0.01 ?g (15 picomoles) of soman-labeled albumin. In conclusion, a high-affinity, polyclonal antibody that specifically recognizes soman adducts on tyrosine in a variety of proteins has been produced. Such an antibody could be useful for identifying secondary targets of soman toxicity. PMID:23469927

Li, Bin; Duysen, Ellen G.; Froment, Marie-Thérčse; Masson, Patrick; Nachon, Florian; Jiang, Wei; Schopfer, Lawrence M.; Thiele, Geoffrey M.; Klassen, Lynell W.; Cashman, John; Williams, Gareth R.; Lockridge, Oksana

2013-01-01

397

Risk assessment in patients with antiphospholipid antibodies.  

PubMed

Antiphospholipid syndrome (APS) is an autoimmune disorder characterized by the association of antiphospholipid antibodies (aPL) with thrombosis and/or pregnancy loss: classification criteria were defined in the updated international consensus held in Sidney in 2005. Vascular and obstetric manifestations display partially different pathogenetic mechanisms. Thrombosis develop as a result of local procoagulative changes upon triggers influence (second-hit theory). Pregnancy morbidity is thought to be dependent on placental thrombosis and complement activation. The laboratory tests include Lupus Anticoagulant (LA), a functional assay, and anticardiolipin (aCL) and anti-?2-glycoprotein I antibodies detected by solid phase enzyme-linked immunosorbent assay (ELISA). The LA testing is relatively standardized while there's still significant interlaboratory discrepancy in ELISA tests. Current APS criteria are under discussion: since for vascular and obstetric APS, different pathogenetic mechanisms have been shown, some criteria variation could also be contemplated. What is the weight of aPL antibodies in provoking thrombosis and which contribution could be expected from aPL per se is debated. As thrombosis is generally considered to be multi-factorial, each case needs a risk-stratified approach. Any primary prophylaxis, intensity and duration of secondary prophylaxis should take into account aPL profile, other cardiovascular risk factors and systemic autoimmune diseases associated. We look forward to the publication of recommendations of the leading experts in the field, developed during the recent 14th International Congress in Rio de Janeiro, Brazil. PMID:24316917

Bertero, M; Kuzenko, A

2013-12-01

398

Antibody-Based Therapies in Multiple Myeloma  

PubMed Central

The unmet need for improved multiple myeloma (MM) therapy has stimulated clinical development of monoclonal antibodies (mAbs) targeting either MM cells or cells of the bone marrow (BM) microenvironment. In contrast to small-molecule inhibitors, therapeutic mAbs present the potential to specifically target tumor cells and directly induce an immune response to lyse tumor cells. Unique immune-effector mechanisms are only triggered by therapeutic mAbs but not by small molecule targeting agents. Although therapeutic murine mAbs or chimeric mAbs can cause immunogenicity, the advancement of genetic recombination for humanizing rodent mAbs has allowed large-scale production and designation of mAbs with better affinities, efficient selection, decreasing immunogenicity, and improved effector functions. These advancements of antibody engineering technologies have largely overcome the critical obstacle of antibody immunogenicity and enabled the development and subsequent Food and Drug Administration (FDA) approval of therapeutic Abs for cancer and other diseases. PMID:22046572

Tai, Yu-Tzu; Anderson, Kenneth C.

2011-01-01

399

Trial Watch: Monoclonal antibodies in cancer therapy.  

PubMed

Since the advent of hybridoma technology, dating back to 1975, monoclonal antibodies have become an irreplaceable diagnostic and therapeutic tool for a wide array of human diseases. During the last 15 years, several monoclonal antibodies (mAbs) have been approved by FDA for cancer therapy. These mAbs are designed to (1) activate the immune system against tumor cells, (2) inhibit cancer cell-intrinsic signaling pathways, (3) bring toxins in the close proximity of cancer cells, or (4) interfere with the tumor-stroma interaction. More recently, major efforts have been made for the development of immunostimulatory mAbs that either enhance cancer-directed immune responses or limit tumor- (or therapy-) driven immunosuppression. Some of these antibodies, which are thought to facilitate tumor eradication by initiating or sustaining a tumor-specific immune response, have already entered clinical trials. In this Trial Watch, we will review and discuss the clinical progress of the most important mAbs that are have entered clinical trials after January 2008. PMID:22720209

Galluzzi, Lorenzo; Vacchelli, Erika; Fridman, Wolf Hervé; Galon, Jerome; Sautčs-Fridman, Catherine; Tartour, Eric; Zucman-Rossi, Jessica; Zitvogel, Laurence; Kroemer, Guido

2012-01-01

400

Antinuclear antibodies: two-step detection strategy.  

PubMed

In order to evaluate the performance of the chemiluminescent immunoassay (CLIA) in antinuclear antibodies (ANA) testing, using indirect fluorescent antibody (IFA) on HEp-2 cells as a standard, 209 samples were studied from September to October/2010. The tests were conducted according to the procedures recommended by the manufacturers of the reagents. The interpretation of the IFA results was done according to the Brazilian standards. The charts of patients showing different results between the two techniques were analyzed. The CLIA efficiency was 89%, with a sensitivity of 65%, and a specificity of 94.7%. Nine (4.3%) false-positive and 14 (6.7%) false-negative results were detected. Of these, 13 (93%) represented no risk for the diagnosis and therapeutic management of the patients. The CLIA methodology reduced the need for the IFA manual technique by 77% (160/209). The ANA screening test proved to be a fast and acceptable methodology in the studied population. We established the following criteria for the introduction of an automated ANA screening: (1) Positive results must be confirmed by IFA to characterize the pattern and titer of the antibody. (2) Negative results are issued with a notice to request a new test by IFA when the clinical suspicion of autoimmune disease persists. PMID:24131388

Callado, Maria Roseli Monteiro; de Alencar Barroso, Maria Nancy; Alves, Vania Maria; de Lima Abreu, Maria Arenilda; Muniz, Lívia M Mesquita Mororó; Lima, José Rubens Costa

2014-01-01

401

Monoclonal antibodies based on hybridoma technology.  

PubMed

Based on the size and scope of the present global market for medicine, monoclonal antibodies (mAbs) have a very promising future, with applications for cancers through autoimmune ailments to infectious disease. Since mAbs recognize only their target antigens and not other unrelated proteins, pinpoint medical treatment is possible. Global demand is dramatically expanding. Hybridoma technology, which allows production of mAbs directed against antigens of interest is therefore privileged. However, there are some pivotal points for further development to generate therapeutic antibodies. One is selective generation of human mAbs. Employment of transgenic mice producing human antibodies would overcome this problem. Another focus is recognition sites and conformational epitopes in antigens may be just as important as linear epitopes, especially when membrane proteins such as receptors are targeted. Recognition of intact structures is of critical importance for medical purposes. In this review, we describe patent related information for therapeutic mAbs based on hybridoma technology and also discuss new advances in hybridoma technology that facilitate selective production of stereospecific mAbs. PMID:24237029

Yagami, Hisanori; Kato, Hiroshi; Tsumoto, Kanta; Tomita, Masahiro

2013-03-01

402

Monoclonal Antibodies Against Xenopus Greatwall Kinase  

PubMed Central

Mitosis is known to be regulated by protein kinases, including MPF, Plk1, Aurora kinases, and so on, which become active in M-phase and phosphorylate a wide range of substrates to control multiple aspects of mitotic entry, progression, and exit. Mechanistic investigations of these kinases not only provide key insights into cell cycle regulation, but also hold great promise for cancer therapy. Recent studies, largely in Xenopus, characterized a new mitotic kinase named Greatwall (Gwl) that plays essential roles in both mitotic entry and maintenance. In this study, we generated a panel of mouse monoclonal antibodies (MAbs) specific for Xenopus Gwl and characterized these antibodies for their utility in immunoblotting, immunoprecipitation, and immunodepletion in Xenopus egg extracts. Importantly, we generated an MAb that is capable of neutralizing endogenous Gwl. The addition of this antibody into M-phase extracts results in loss of mitotic phosphorylation of Gwl, Plk1, and Cdk1 substrates. These results illustrate a new tool to study loss-of-function of Gwl, and support its essential role in mitosis. Finally, we demonstrated the usefulness of the MAb against human Gwl/MASTL. PMID:22008075

Wang, Ling; Fisher, Laura A.; Wahl, James K.

2011-01-01

403

Monoclonal antibodies against Xenopus greatwall kinase.  

PubMed

Mitosis is known to be regulated by protein kinases, including MPF, Plk1, Aurora kinases, and so on, which become active in M-phase and phosphorylate a wide range of substrates to control multiple aspects of mitotic entry, progression, and exit. Mechanistic investigations of these kinases not only provide key insights into cell cycle regulation, but also hold great promise for cancer therapy. Recent studies, largely in Xenopus, characterized a new mitotic kinase named Greatwall (Gwl) that plays essential roles in both mitotic entry and maintenance. In this study, we generated a panel of mouse monoclonal antibodies (MAbs) specific for Xenopus Gwl and characterized these antibodies for their utility in immunoblotting, immunoprecipitation, and immunodepletion in Xenopus egg extracts. Importantly, we generated an MAb that is capable of neutralizing endogenous Gwl. The addition of this antibody into M-phase extracts results in loss of mitotic phosphorylation of Gwl, Plk1, and Cdk1 substrates. These results illustrate a new tool to study loss-of-function of Gwl, and support its essential role in mitosis. Finally, we demonstrated the usefulness of the MAb against human Gwl/MASTL. PMID:22008075

Wang, Ling; Fisher, Laura A; Wahl, James K; Peng, Aimin

2011-10-01

404

Antibodies to Orientia tsutsugamushi in Thai soldiers.  

PubMed

Thai soldiers who were conscripted, Royal Thai Army forces, professional Border Patrol Police, or local militia (Thai Rangers) located in any of seven provinces of Thailand were bled in April and again, four months later, in July 1989. In 1991, soldiers from five different locations in southern Thailand were bled once, in July. Serum samples were tested by indirect fluorescent antibody assay for antibody to Orientia (formerly Rickettsia) tsutsugamushi, etiologic agent of scrub typhus, with any titer > or = 1:50 considered positive. Prior to field exercises, prevalence of antibody varied significantly between different types of units, ranging between 18.6% for Thai Rangers and 6.8% for the Royal Thai Army. The April prevalence, July prevalence, and incidence varied significantly by province in 1989, with highest incidence being 14.5% in Kanchanaburi and the lowest 0% in Utraladit. The prevalence in southern Thailand in 1991 varied between 1.6% and 6.8%. The data demonstrate that O. tsutsugamushi is widely distributed in Thailand and that military activity consisting of field exercises that simulate combat conditions significantly expose soldiers to infection. PMID:8940989

Eamsila, C; Singsawat, P; Duangvaraporn, A; Strickman, D

1996-11-01

405

Antibody neutralization of retargeted measles viruses.  

PubMed

The measles virus (MV) vaccine lineage is a promising oncolytic but prior exposure to the measles vaccine or wild-type MV strains limits treatment utility due to the presence of anti-measles antibodies. MV entry can be redirected by displaying a polypeptide ligand on the Hemagglutinin (H) C-terminus. We hypothesized that retargeted MV would escape neutralization by monoclonal antibodies (mAbs) recognizing the H receptor-binding surface and be less susceptible to neutralization by human antisera. Using chimeric H proteins, with and without mutations that ablate MV receptor binding, we show that retargeted MVs escape mAbs that target the H receptor-binding surface by virtue of mutations that ablate infection via SLAM and CD46. However, C-terminally displayed domains do not mediate virus entry in the presence of human antibodies that bind to the underlying H domain. In conclusion, utility of retargeted oncolytic measles viruses does not extend to evasion of human serum neutralization. PMID:24725950

Lech, Patrycja J; Pappoe, Roland; Nakamura, Takafumi; Tobin, Gregory J; Nara, Peter L; Russell, Stephen J

2014-04-01

406

Synthetic haptens as probes of antibody response and immunorecognition.  

PubMed

The molecular forces that bind antibody to antigen have long fascinated chemists. The use of synthetic haptens to study immunochemical phenomena can be traced back to the classic work of Karl Lansteiner. His utilization of small-molecule-protein conjugates first demonstrated the shape-selective nature of antibody binding. Later work by Linus Pauling and David Pressman employed multivalent, synthetic ligands to establish the bivalent nature of antibodies and explain the nature of immunoprecipitation. Fluorescent probes such as dansyl, fluorescein, and Ru(bpy)(2+)(3) have been used to study affinity maturation, quantify antibody affinities, and investigate polyclonal antibody heterogeneity. Finally, X-ray crystallography has yielded a molecular picture of how antibodies exercise intermolecular forces (e.g., charge-charge interactions, H-bonding, and Van der Waals) to bind haptens. Studies inspired by Landsteiner's original work continue to play an important role in fields ranging from immunodiagnostics to catalytic antibodies. PMID:10694458

Shreder, K

2000-03-01

407

Antibody vs. HIV in a clash of evolutionary titans.  

PubMed

HIV has evolved many strategies to avoid neutralizing antibody responses, particularly to conserved regions on the external glycoprotein spikes of the virus. Nevertheless, a small number of antibodies have been evolved by the human immune system to recognize conserved parts of the glycoproteins, and therefore, have broadly neutralizing cross-strain activities. These antibodies constitute important tools in the quest to design immunogens that can elicit broadly neutralizing antibodies in humans and hence contribute to an effective HIV vaccine. Crystallographic analyses of the antibodies, in many cases in an antigen-complexed form, have revealed novel and, in some instances, remarkable structural adaptations to attain virus recognition. Antibodies, like HIV, can evolve relatively rapidly through mutation and selection. It seems that the structures of these broadly neutralizing antibodies bear witness to a heroic struggle between two titans of rapid evolution. PMID:16219699

Burton, Dennis R; Stanfield, Robyn L; Wilson, Ian A

2005-10-18

408

Synthesis of antibodies, including antiviral antibodies, in the knee joints of patients with arthritis.  

PubMed Central

Serum and synovial fluids from 16 patients with seronegative arthritis and eight with rheumatoid arthritis were studied for immunoglobulin levels and for antibody levels to five viruses. When allowances were made for the distribution of immunoglobulins between serum and synovial fluid there was evidence that in several patients antibody to one or more viruses was synthesised locally in the joint. IgG and especially IgM were present in greatly increased amounts in arthritic joints compared with normal joints. On the basis of serum/synovial fluid ratios inflammation and local immunoglobulin synthesis are discussed as possible causes. These results are compared with antiviral antibody and immunoglobulin ratios observed in the serum and cerebrospinal fluid of patients with multiple sclerosis. PMID:4062387

Mims, C A; Stokes, A; Grahame, R

1985-01-01

409

Anti-idiotypic antibodies induce neutralizing antibodies to bovine herpesvirus 1.  

PubMed Central

A neutralizing murine monoclonal antibody (mAb) of the IgG2a isotype (MM-113), specific for bovine herpesvirus 1 (BHV-1) glycoprotein gIV, was used to develop anti-idiotypic antibodies (anti-Id) in a calf. The bovine anti-Id were isolated from the serum of the immunized calf by affinity chromatography on an MM-113-Sepharose column, followed by repeated adsorption on a murine IgG2a column. The anti-Id thus obtained specifically reacted with MM-113, but not with isotype-matched controls. They also inhibited the binding of MM-113 to BHV-1 in a concentration-dependent manner. Mice immunized with the anti-Id produced neutralizing antibodies to BHV-1. The anti-Id bound to cells permissive to BHV-1 in a cell-binding radioimmunoassay (RIA). PMID:2165998

Srikumaran, S; Onisk, D V; Borca, M V; Nataraj, C; Zamb, T J

1990-01-01

410

Transplacental transfer of immune antibodies in the mouse demonstrated by antibody labeled in vivo with tritium  

E-print Network

. , Eisen, H. N. , Siegal, M. , Fitzgerald, P. J, , Sherman, B. , and Silverstein, A. : The Zone of Localization of Anti- bodies. J. Immunol. , 65, (1950): 559-563. 24. Prouvost-Danon, A. , and Binaghi, R. : Reaginic Antibody in Adult and Young Mice. Int.... , Eisen, H. N. , Siegal, M. , Fitzgerald, P. J, , Sherman, B. , and Silverstein, A. : The Zone of Localization of Anti- bodies. J. Immunol. , 65, (1950): 559-563. 24. Prouvost-Danon, A. , and Binaghi, R. : Reaginic Antibody in Adult and Young Mice. Int...

McKinney, Hubert Eugene

2012-06-07

411

Altered Antibody Profiles against Common Infectious Agents in Chronic Disease  

PubMed Central

Despite the important diagnostic value of evaluating antibody responses to individual human pathogens, antibody profiles against multiple infectious agents have not been used to explore health and disease mainly for technical reasons.  We hypothesized that the interplay between infection and chronic disease might be revealed by profiling antibodies against multiple agents. Here, the levels of antibodies against a panel of 13 common infectious agents were evaluated with the quantitative Luciferase Immunoprecipitation Systems (LIPS) in patients from three disease cohorts including those with pathogenic anti-interferon-? autoantibodies (IFN-? AAB), HIV and Sjögren’s syndrome (SjS) to determine if their antibody profiles differed from control subjects.  The IFN-? AAB patients compared to controls demonstrated statistically higher levels of antibodies against VZV (p=0.0003), EBV (p=0.002), CMV (p=0.003), and C. albicans (p=0.03), but lower antibody levels against poliovirus (p=0.04). Comparison of HIV patients with blood donor controls revealed that the patients had higher levels of antibodies against CMV (p=0.0008), HSV-2 (p=0.0008), EBV (p=0.001), and C. albicans (p=0.01), but showed decreased levels of antibodies against coxsackievirus B4 (p=0.0008), poliovirus (p=0.0005),   and HHV-6B (p=0.002). Lastly, SjS patients had higher levels of anti-EBV antibodies (p=0.03), but lower antibody levels against several enteroviruses including a newly identified picornavirus, HCoSV-A (p=0.004), coxsackievirus B4 (p=0.04), and poliovirus (p=0.02). For the IFN-? AAB and HIV cohorts, principal component analysis revealed unique antibody clusters that showed the potential to discriminate patients from controls.  The results suggest that antibody profiles against these and likely other common infectious agents may yield insight into the interplay between exposure to infectious agents, dysbiosis, adaptive immunity and disease activity. PMID:24312567

Burbelo, Peter D.; Ching, Kathryn H.; Morse, Caryn G.; Alevizos, Ilias; Bayat, Ahmad; Cohen, Jeffrey I.; Ali, Mir A.; Kapoor, Amit; Browne, Sarah K.; Holland, Steven M.; Kovacs, Joseph A.; Iadarola, Michael J.

2013-01-01

412

Implications of neutralising antibodies on therapeutic efficacy.  

PubMed

All biopharmaceutical preparations are potentially immunogenic and need to be evaluated for the risk of development of neutralising antibodies (NAbs). In the case of interferon-beta preparations for the treatment of multiple sclerosis, persistently high-titres of NAbs are detectable in up to one-third of patients depending on the preparation used. In contrast, treatment with glatiramer acetate is not associated with the development of such antibodies, since its mechanism of action does not involve binding and activation of a specific receptor. The development of NAbs to interferon-beta abrogates biological activity in the short term, although the loss of clinical benefit usually only becomes manifest a year or more after antibodies are first detected. This loss of clinical benefit is apparent as a recrudescence of disease activity visible on magnetic resonance imaging, an increase in the frequency of relapses and a progression of neurological disability. Loss of biological activity can be detected very early after the appearance of NAbs using the MxA test, which can be used as a prognostic biological marker for probable future treatment failure. Current European guidelines recommend that all patients treated with an interferon-beta should be monitored systematically for the appearance of NAbs and that patients who develop persistent NAbs should discontinue their interferon-beta treatment. An option for the patients who have experienced less than two relapses in the previous year is to switch to treatment with glatiramer acetate. Such a strategy offers the best guarantee of providing good control of disease activity and optimal allocation of healthcare resources. PMID:19200862

Bertolotto, Antonio

2009-02-01

413

Prevalence of antipituitary antibodies in acromegaly.  

PubMed

Acromegaly is a rare disorder due to an excessive production of growth hormone (GH), typically caused by a GH-secreting pituitary adenoma. Anti-pituitary antibodies (APAs) are often seen in patients with different kinds of pituitary pathologies. Because GH has been proposed as a possible antigen recognized by such antibodies, the prevalence of APAs may be higher in conditions characterized by excessive GH secretion. The primary aim of this study was to compare the prevalence of APAs in patients with acromegaly and in controls with other types of pituitary tumors and healthy subjects. Secondary aim was to characterize the pituitary cells targeted by the APAs. Thirty eight acromegaly patients and 215 controls, including 38 patients with prolactinomas, 64 with non-functioning pituitary adenomas (NFPA), and 113 healthy subjects were enrolled in the study. All subjects were tested for APAs using indirect immunofluorescence. Target cells recognized by APAs were identified by double staining immunofluorescence. APAs were significantly more prevalent in acromegaly cases than in healthy controls (10.5% vs. 1.8%, P < 0.05). This prevalence was similar to that found in patients with prolactinomas (7.9%) and NFPA (12.5%). Among APAs-positive subjects, antibodies recognizing somatotrope cells were more common in acromegaly cases than in healthy controls (3/4 vs. 0/113, P < 0.0001), but had similar frequencies in NFPA (2/8) and prolactinomas (1/3). APAs are more frequently found in patients with pituitary adenomas than healthy subjects, with no significant difference among the tumor types studied. GH-secreting cells could represent a target of the autoimmune response. PMID:22002711

Guaraldi, Federica; Caturegli, Patrizio; Salvatori, Roberto

2012-12-01

414

Phylogenetic study of transcortin using monoclonal antibodies.  

PubMed

We produced monoclonal antibodies that recognise three distinct epitopes of human transcortin. These epitopes are present on transcortin of humans with normal and altered transcortin levels, as well as on a variant with lower affinity for cortisol. One epitope is present on transcortin of Old World Monkeys and apes, the others are only present on transcortin of apes. The epitopes are not present on transcortin of other species. These results indicate that human transcortin contains a highly evolved and a more conserved part. PMID:2428359

Faict, D; De Moor, P

1986-08-14

415

[Natalizumab: An Antibody Targeting ?4-Integrin].  

PubMed

Abstract Multiple sclerosis (MS) is a demyelinating disease of the central nervous system (CNS), in which inflammation develops upon leukocyte invasion into the CNS via the blood-brain barrier. ?4-Integrin is an important cell adhesion molecule involved in the penetration process. Natalizumab is an ?4-integrin-targeting monoclonal antibody that was recently approved for use as a disease-modifying therapy for MS in Japan. In this article, the mechanism of action, efficacy, and side effects of natalizumab will be reviewed. PMID:25296869

Nakahara, Jin

2014-10-01

416

Novel antibody switching defects in human patients  

PubMed Central

Hyper-IgM syndrome (HIGM) is a primary immunodeficiency characterized by normal to elevated serum levels of IgM and low levels or the absence of IgG, IgA, and IgE. A new study AID expression in nonlymphoid cells (see related article on pages 136–142) characterizes HIGM type 4, a previously undocumented defect in antibody gene diversification caused by a selective block in class-switch recombination, providing significant insight towards understanding HIGM immunodeficiencies. PMID:12840053

Manis, John P.; Alt, Frederick W.

2003-01-01

417

Assays of thyroid-stimulating antibody  

SciTech Connect

A comparison is presented of the two major assay methods of thyroid-stimulating antibody (TSAb) of Graves' disease. The basic procedures involve: (1) some index of thyroid stimulation, usually in vitro, using TSAb to indicate its activity; and (2) indirect recognition by assessment of the inhibition of binding of radioiodinated thyrotropin (TSH) to a preparation of its receptor, i.e., TSH-binding inhibition or TBI. There is potential for misinterpretation of data acquired by testing patients' sera by one or the other basic procedure.

McKenzie, J.M.; Zakarija, M.

1985-01-01

418

Spectrum of antibodies to reproductive hormones in threatened abortion.  

PubMed

The spectrum of antibodies to reproductive hormones and the diagnostic significance of their measurements in threatened abortion during trimester I were studied. Enhanced production of antibodies to hormones was detected by ELISA in patients with threatened abortion (N=44) in comparison with women with normal gestation (N=30). These antibodies were detected more often than antiphospholipid antibodies (p<0.05). Antibodies to chorionic gonadotropin (IgM, IgG) and gonadotropin-releasing hormone (IgG) were associated with threatened abortion. According to ROC analysis, their measurements were diagnostically significant in this pathology (AUC>0.8). Subclasses IgG1 and IgG2 predominated among IgG to chorionic gonadotropin. Presumably, antibodies to chorionic gonadotropin and gonadotropin-releasing hormone could serve as independent factors of threatened abortion risk during trimester I. PMID:25348563

Menzhinskaya, I V; Van'ko, L V; Kiryushchenkov, P A; Tambovtseva, M A; Kashentseva, M M; Sukhikh, G T

2014-10-01

419

Generation of neutralising antibodies against porcine endogenous retroviruses (PERVs)  

SciTech Connect

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.

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

2011-03-01

420

Antibody-mediated prevention of Fusarium mycotoxins in the field.  

PubMed

Fusarium mycotoxins directly accumulated in grains during the infection of wheat and other cereal crops by Fusarium head blight (FHB) pathogens are detrimental to humans and domesticated animals. Prevention of the mycotoxins via the development of FHB-resistant varieties has been a challenge due to the scarcity of natural resistance against FHB pathogens. Various antibodies specific to Fusarium fungi and mycotoxins are widely used in immunoassays and antibody-mediated resistance in planta against Fusarium pathogens has been demonstrated. Antibodies fused to antifungal proteins have been shown to confer a very significantly enhanced Fusarium resistance in transgenic plants. Thus, antibody fusions hold great promise as an effective tool for the prevention of mycotoxin contaminations in cereal grains. This review highlights the utilization of protective antibodies derived from phage display to increase endogenous resistance of wheat to FHB pathogens and consequently to reduce mycotoxins in field. The role played by Fusarium-specific antibody in the resistance is also discussed. PMID:19325726

Hu, Zu-Quan; Li, He-Ping; Zhang, Jing-Bo; Glinka, Elena; Liao, Yu-Cai

2008-10-01

421

Cerebrospinal Fluid Aquaporin-4 Antibody Levels in Neuromyelitis Optica Attacks  

PubMed Central

To elucidate immunopathogenetic roles of aquaporin-4 antibodies in the cerebrospinal fluid (CSF) of neuromyelitis optica spectrum disorders (NMOSD), we analyzed aquaporin-4 antibody titers, cellular and inflammatory markers in the CSF collected from 11 aquaporin-4 antibody seropositive patients. The CSF aquaporin-4 antibody levels during attacks (but not in sera) closely correlated with pleocytosis, inflammatory cytokines including interleukin-6 that can regulate antibody-producing plasmablasts, and glial fibrillary acidic protein levels in the CSF. The amount of aquaporin-4 antibodies present in the central nervous system may have therapeutic implications, as it is associated with astrocyte injury and inflammatory responses during NMOSD attacks. Ann Neurol 2014;76:305–309 PMID:24977390

Sato, Douglas Kazutoshi; Callegaro, Dagoberto; de Haidar Jorge, Frederico M; Nakashima, Ichiro; Nishiyama, Shuhei; Takahashi, Toshiyuki; Simm, Renata Faria; Apostolos-Pereira, Samira Luisa; Misu, Tatsuro; Steinman, Lawrence; Aoki, Masashi; Fujihara, Kazuo

2014-01-01

422

In Vitro Antibody Affinity Maturation Targeting Germline Hotspots  

PubMed Central

Affinity-matured antibodies can exhibit increased biological efficacy. Regardless of whether an antibody is isolated from a hybridoma or a human Fv phage library, the antibody affinity for its target may need improvement for therapeutic applications. An increased affinity may allow for a reduced dosage of a therapeutic antibody; toxic side effects may also be reduced. In the immune system, affinity maturation is a process involving somatic hypermutations in B cells. Therefore, germline hotspot residues are most likely to have a major impact on antibody affinity. Here, we describe procedures for germline hotspot mutagenesis with an emphasis on strategies for randomizing hotspots with PCR and phage display, using as an example the anti-CD22 monoclonal antibody. PMID:19252855

Ho, Mitchell; Pastan, Ira

2012-01-01

423

Minimal requirements for antiphospholipid antibodies ELISAs proposed by the European Forum on antiphospholipid antibodies  

Microsoft Academic Search

Antiphospholipid ELISAs are part of the Antiphospholipid Antibodies Syndrome classification criteria, having the same diagnostic value as lupus anticoagulant. However, sometimes their results appear scarcely meaningful especially when wide metanalyses studies are performed, probably because of their well-known inter-laboratory variability. The application of a common protocol was shown to improve the test reproducibility, but this observation did not have any

Angela Tincani; Flavio Allegri; Genesio Balestrieri; Guido Reber; Marielle Sanmarco; Pierluigi Meroni; Marie-Claire Boffa

2004-01-01

424

Antibodies have rapidly become a clinically important drug class: more than 25 antibodies are approved for  

E-print Network

are approved for humantherapyandmorethan240antibodiesarecurrently in clinical development worldwide for a wide has led to a major commercial impact, with rapidly growing annual sales that exceeded US$27 billion display libraries expressing human antibody fragments or transgenic mice engi- neered with human

Cai, Long

425

An efficient method for isolating antibody fragments against small peptides by antibody phage display.  

PubMed

We generated monoclonal scFv (single chain variable fragment) antibodies from an antibody phage display library towards three small synthetic peptides derived from the sequence of ?s1-casein. Key difficulties for selection of scFv-phages against small peptides were addressed. Small peptides do not always bind efficiently to passive adsorption surfaces, and we developed a simple method to quantify the binding capacity of surfaces with the peptides. Background binding (the binding of scFvs to the background matrix) is an obstacle for successful selection, and we evaluated two methods that drastically reduced the background binding. An optimized method therefore enabled a panning procedure where the specific (peptide binding) scFv-phages were always dominant. Using 15-mer peptides immobilized on Nunc Immobilizer Streptavidin plates, we successfully generated scFvs specifically against them. The scFvs were sequenced and characterized, and specificity was characterized by ELISA. The methods developed in this study are universally applicable for antibody phage display to efficiently produce antibody fragments against small peptides. PMID:20615194

Duan, Zhi; Siegumfeldt, Henrik

2010-11-01

426

Antibody-guided irradiation of advanced ovarian cancer with intraperitoneal monoclonal antibodies  

SciTech Connect

Twenty-four patients with persistent epithelial ovarian cancer after chemotherapy with or without external beam irradiation, were treated with intraperitoneally administered /sup 131/I-labeled monoclonal antibodies HMFG1, HMFG2, AUA1, H17E2, directed against tumor-associated antigens. Acute side effects were mild abdominal pain, pyrexia, diarrhea, and moderate reversible pancytopenia. One patient developed a subphrenic abscess requiring surgical drainage. Eight patients with large volume disease, ie, greater than 2 cm tumor diameter, did not respond to antibody-guided irradiation and died of progressive disease within 9 months of treatment. Sixteen patients had small-volume (less than 2 cm) disease at the time of treatment with radiolabeled antibody. Seven patients failed to respond, and of nine initial responders, four patients remain alive and free from disease 6 months to 3 years from treatment. Analysis of the data on relapse indicated that doses greater than 140 mCi were more effective than lower doses. We conclude that the intraperitoneal administration of 140 mCi or more of /sup 131/I-labeled tumor-associated monoclonal antibodies represents a new and potentially effective form of therapy for patients with small-volume stage III ovarian cancer.

Epenetos, A.A.; Munro, A.J.; Stewart, S.; Rampling, R.; Lambert, H.E.; McKenzie, C.G.; Soutter, P.; Rahemtulla, A.; Hooker, G.; Sivolapenko, G.B.

1987-12-01

427

Proper application of antibodies for immunohistochemical detection: antibody crimes and how to prevent them.  

PubMed

For several decades antibodies raised against specific proteins, peptides, or peptide epitopes have proven to be versatile and very powerful tools to demonstrate molecular identity in cells and tissues. New techniques of immunohistochemistry and immunofluorescence have improved both the optical resolution of such protein identification as well as its sensitivity, particularly through the use of amplification methodology. However, this improved sensitivity has also increased the risks of false-positive and false-negative staining and thereby raised the necessity for proper and adequate controls. In this review, the authors draw on many years of experience to illuminate many of the more common errors and problematic issues in immunohistochemistry, and how these may be avoided. A key factor in all of this is that techniques need to be properly documented and especially antibodies and procedures must be adequately described. Antibodies are a valuable and shared resource within the scientific community; it is essential therefore that mistakes involving antibodies and their controls are not perpetuated through inadequate reporting in the literature. PMID:24428532

Ivell, Richard; Teerds, Katja; Hoffman, Gloria E

2014-03-01

428

Assessment of a fluorescent antibody test for the detection of antibodies against epizootic bovine abortion.  

PubMed

The current study was directed at developing and validating an indirect fluorescent antibody test (IFAT) capable of detecting antibodies specific for the agent of epizootic bovine abortion (aoEBA). Sensitivity and specificity was determined by comparing antibody titers from 114 fetuses infected with aoEBA with 68 fetuses diagnosed with alternate infectious etiologies. Data established specificity at 100% and sensitivity at 94.7% when cutoff criteria for a positive test were assigned at a titer of ?1,000. Potential cross-reactivity was noted in samples from 3 fetuses with antibody titers of 10 or100; all were infected with Gram-positive organisms. The remaining 65 fetuses infected with microbes other than aoEBA, and an additional 12 negative reference sera, did not have detectable titers. The IFAT-based serology assay is rapid, reproducible, and unaffected by fluid color or opacity. Total fetal immunoglobulin (Ig)G was also evaluated as an aid for diagnosing EBA. Significantly higher concentrations of IgG were identified in fetuses infected with aoEBA as compared to those with alternate infectious etiologies. The presence of IgG is a sensitive indicator of EBA and increases the specificity of FAT-based serologic diagnosis when titers are 10 or 100. Taken together, serology and IgG analyses suggest that the incidence of EBA may be underestimated. PMID:25139792

Blanchard, Myra T; Anderson, Mark L; Hoar, Bruce R; Pires, Alda F A; Blanchard, Patricia C; Yeargan, Bret V; Teglas, Mike B; Belshaw, Margaret; Stott, Jeffery L

2014-09-01

429

Identification of Pigment Cell Antigens Defined by Vitiligo Antibodies  

Microsoft Academic Search

Patients with vitiligo have circulating antibodies to pigment cells. To characterize this response further and to identify the antigens defined by vitiligo antibodies, sera of 23 patients with vitiligo and 22 patients with unrelated conditions were analyzed by immunoprecipitation and SDS-PAGE analysis of 125I-labeled cell antigens on pigment and control cells. Antibodies to pigment cell antigens were present in 18

Jian Cui; Ronald Harning; Milagros Henn; Jean-Claude Bystryn

1992-01-01

430

Effect of beta propiolactone on specific antibody measurements by ELISA.  

PubMed Central

Serum samples from 30 HIV seronegative patients were treated with beta propiolactone (BPL) to determine whether BPL interfaces with ELISA for specific antibodies against protein and carbohydrate antigens. BPL had no discernible effect on specific antibody measurements by ELISA. With the measuring need for specific antibody measurements in the management of HIV seropositive patients, it is reassuring that this laboratory safety measure does not impair the reliability of results. PMID:8496395

Agbarakwe, A E; Misbah, S A; Griffiths, H; Chapel, H M

1993-01-01

431

Effect of anti-lymphocyte serum on natural antibody  

PubMed Central

Horse anti-dog lymphocyte serum was found to suppress the immune response of dogs normally resulting from the injection of Salmonella typhi vaccine. Normal antibody levels of the dogs against Shigella dysenteriae were unaffected, however, by the anti-lymphocyte serum. The results suggest, therefore, that the natural antibodies may arise by a population of cells or by mechanisms different from those which produce antibodies as a result of deliberate antigenic stimulation. PMID:4867939

Muschel, L. H.; Gustafson, Linda; Atai, M.

1968-01-01

432

Application for Antigen—Antibody Sensor Using Carbon Nanotubes  

NASA Astrophysics Data System (ADS)

An antigen-antibody sensor consisting of a silicon-based chip contains a mixture of poly [ethylene glycol] (PEG)-grafted carbon nanotubes (PEG-CNTs) and CNTs modified with an antigen by using plasma ion irradiation (plasma activation) method, is developed. According to result of experiments, impedance increased due to antigen/antibody reaction. The results indicate that this antigen-antibody sensor could react until 10 g/ml antigen density.

Funada, Yuichiro; Hirata, Takamichi; Akiya, Masahiro

433

Absence of antiphospholipid\\/co-factor antibodies in Takayasu Arteritis  

Microsoft Academic Search

There are anecdotal reports and small series describing the presence of anticardiolipin antibodies in patients with Takayasu Arteritis. This communication describes a systematic study searching for non-organ specific autoantibodies which includes antinuclear antibodies, anticardiolipin and anti-?2 GP1 antibodies in a cohort of 28 Mexicans with angiographic definitive diagnostic of Takayasu Arteritis. Material and methods: Twenty-eight consecutive patients, who fulfilled classification

Arnulfo Nava; Jean Luc Senécal; José Luis Bańales; Ives Raymond; Pedro A. Reyes

2000-01-01

434

Systemic Persistence of Homologous Guinea Pig Skin-Sensitizing Antibodies  

Microsoft Academic Search

Homologous guinea pig skin-sensitizing antibodies of the IgG1 class (as characterized by heat stability and persistence in skin for 7 days) were shown to persist systemically for 28–35 days. Persistence was shown by the anaphylaxis induced in guinea pigs following the intracardiac administration of the antibody and subsequent antigenic challenge by aerosol. Skin-sensitizing antibody of the IgE class, characterized by

Herbert Megel; Donald L. Denney

1979-01-01

435

Immunoprophylaxis in fish by injection of mouse antibody genes  

Microsoft Academic Search

Antibodies are a crucial part of the body's specific defense against infectious diseases and have considerable potential as therapeutic and prophylactic agents in humans and animals. The development of recombinant single-chain antibodies allows a genetic application strategy for prevention of infectious diseases. To test this in a fish model, a gene construct encoding a neutralizing single-chain antibody to the fish-pathogenic

Pauline M. Cupit; Katja Einer-Jensen; Ellen Lorenzen; Peter Ahrens; Christopher J. Secombes; Charles Cunningham; Niels Lorenzen

2000-01-01

436

Skin sensitizing antibody in experimental infections with Metastrongylus spp. (Nematoda)  

PubMed Central

Homologous passive cutaneous anaphylactic tests in guinea-pigs revealed the production of skin sensitizing antibody by Metastrongylus spp. infection. Skin sensitizing antibody was detected by immediate wheal and flare reaction in an infected pig. The pig skin sensitizing antibody failed to elicit a Prausnitz—Kustner (PK) reaction in an uninfected recipient pig, or heterologous passive cutaneous anaphylaxis in guinea-pigs. PMID:5411494

Barratt, M. E. J.; Herbert, I. V.

1970-01-01

437

In vitro assay of reaginic antibodies to horse serum albumin  

PubMed Central

The antibody content of three reaginic sera from patients sensitive to highly purified horse serum albumin have been investigated by measuring the capacity to bind 131I-labelled albumin. No correlation was found between skin sensitizing activity and the content of either heat stable or heat labile antibody, but there was a correlation with the ratio of the concentration in the serum of the two kinds of antibody. ImagesFIG. 2 PMID:6065020

Chan, P. C. Y.; Porter, R. R.

1967-01-01

438

Product-dependent anti-factor VIII antibodies  

PubMed Central

Introduction The development of anti-factor (F)VIII antibodies in hemophilia A (HA) subjects undergoing replacement therapy has been well-documented. The correlation between antibody