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Peptide Epitopes (peptide + epitope)
Selected AbstractsAvailability of autoantigenic epitopes controls phenotype, severity, and penetrance in TCR Tg autoimmune gastritisEUROPEAN JOURNAL OF IMMUNOLOGY, Issue 12 2008Ditza Levin Abstract We examined TCR:MHC/peptide interactions and in vivo epitope availability to explore the Th1- or Th2-like phenotype of autoimmune disease in two TCR Tg mouse models of autoimmune gastritis (AIG). The TCR of strains A23 and A51 recognize distinct IAd -restricted peptides from the gastric parietal cell H/K-ATPase. Both peptides form extremely stable MHC/peptide (MHC/p) complexes. All A23 animals develop a Th1-like aggressive, inflammatory AIG early in life, while A51 mice develop indolent Th2-like AIG at 6,8,wk with incomplete penetrance. A51 T cells were more sensitive than A23 to low doses of soluble antigen and to MHC/p complexes. Staining with IAd/peptide tetramers was only detectable on previously activated T cells from A51. Thus, despite inducing a milder AIG, the A51 TCR displays a higher avidity for its cognate IAd/peptide. Nonetheless, in vivo proliferation of adoptively transferred A51 CFSE-labeled T cells in the gastric lymph node was relatively poor compared with A23 T cells. Also, DC from WT gastric lymph node, presenting processed antigen available in vivo, stimulated proliferation of A23 T cells better than A51. Thus, the autoimmune potential of these TCR in their respective Tg lines is strongly influenced by the availability of the peptide epitope, rather than by differential avidity for their respective MHC/p complexes. [source] Amoebic gill disease resistance is not related to the systemic antibody response of Atlantic salmon, Salmo salar L.JOURNAL OF FISH DISEASES, Issue 1 2010R S Taylor Abstract Amoebic gill disease (AGD) is a proliferative gill tissue response caused by Neoparamoeba perurans and is the main disease affecting Australian marine farmed Atlantic salmon. We have previously proposed that macroscopic gill health (,gill score') trajectories and challenge survival provide evidence of a change in the nature of resistance to AGD. In order to examine whether the apparent development of resistance was because of an adaptive response, serum was sequentially sampled from the same individuals over the first three rounds of natural AGD infection and from survivors of a subsequent non-intervention AGD survival challenge. The systemic immune reaction to ,wildtype'Neoparamoeba sp. was characterized by Western blot analysis and differentiated to putative carbohydrate or peptide epitopes by periodate oxidation reactions. The proportion of seropositive fish increased from 46% to 77% with each AGD round. Antibody response to carbohydrate epitope(s) was immunodominant, occurring in 43,64% of samples. Antibodies that bound peptide epitope were identified in 16% of the challenge survivors. A 1:50 (single-dilution) enzyme-linked immunosorbent assay confirmed a measurable immune titre in 13% of the survivors. There was no evidence that antibodies recognizing wildtype Neoparamoeba provided significant protection against AGD. [source] Viral peptide immunogens: current challenges and opportunitiesJOURNAL OF PEPTIDE SCIENCE, Issue 12 2007Ali Azizi Abstract Synthetic peptide vaccines have potential to control viral infections. Successful experimental models using this approach include the protection of mice against the lethal Sendai virus infection by MHC class I binding CTL peptide epitope. The main benefit of vaccination with peptide epitopes is the ability to minimize the amount and complexity of a well-defined antigen. An appropriate peptide immunogen would also decrease the chance of stimulating a response against self-antigens, thereby providing a safer vaccine by avoiding autoimmunity. In general, the peptide vaccine strategy needs to dissect the specificity of antigen processing, the presence of B-and T-cell epitopes and the MHC restriction of the T-cell responses. This article briefly reviews the implications in the design of peptide vaccines and discusses the various approaches that are applied to improve their immunogenicity. Copyright © 2007 European Peptide Society and John Wiley & Sons, Ltd. [source] Enhancement of Immunogenicity of Jeg3 Cells by Ectopic Expression of HLA-A*0201 and CD80AMERICAN JOURNAL OF REPRODUCTIVE IMMUNOLOGY, Issue 3 2003Serpil Koc Problem: The choriocarcinoma cell line Jeg3 suppresses immunity in vitro by secretion of soluble factors like leukemia inhibitory factor suppressing leukocyte activation. The cells lack expression of classical human leukocyte antigen (HLA)-A and -B alleles but express some HLA-C, and non-classical HLA-G and -E. Upon binding to killing inhibitory receptor on natural killer (NK) cells, HLA-G prevents activation of cytolytic activity. We investigated whether Jeg3 cells are capable of immune stimulation after complementation with classical HLA and T cell costimulatory signal CD80. Method of study: Jeg3 cells were transduced to express HLA-A*0201 and/or CD80. Parental Jeg3 or transfectants Jeg3-A2, Jeg3-CD80 or Jeg3-CD80-A2 were used to stimulate allogeneic resting and activated peripheral blood lymphocytes (PBL). The different cell lines were loaded with a HLA-A2-restricted Epstein-Barr virus (EBV) recall antigen peptide epitope and antigen presenting ability was examined. T cell lines specific for Jeg3 and transfectants were generated from HLA-A2 matched and nonmatched donors and compared for expansion, phenotypes and cytolytic activity. Results: While all Jeg3 cell lines induced only marginal proliferation of resting T cells, phytohemagglutinin (PHA)-activated T cells were stimulated by CD80 or CD80-A2 expressing Jeg3. Only the transfectant Jeg3-CD80-A2 was capable of specific T cell stimulation by EBV recall antigen presentation. T cell lines of HLA-A2 non-matched donors stimulated with the Jeg3 transfectants showed significant expansion only when HLA-A2 and the costimulus CD80 were present. T cells from HLA-A2 positive donors did not expand significantly or differentially. No NK cells grew under any condition. In Jeg3-CD80-A2 stimulated T cells lines CD8+ cells expanded preferentially. These T cells exerted cytolytic activity toward all Jeg3 cell lines. Conclusion: Our data suggest that, in spite of immunosuppressive mechanisms, proliferative and cytolytic T cell responses are induced by Jeg3 cells when classical HLA- and/or costimulatory signals are present on the cells. [source] Identification of naturally processed CD8 T,cell epitopes from prostein, a prostate tissue-specific vaccine candidateEUROPEAN JOURNAL OF IMMUNOLOGY, Issue 4 2004Rachel Abstract Prostein is a prostate tissue-specific protein that is uniquely and abundantly expressed in normal and cancerous prostate tissues. Due to this expression profile, we examined the immunogenicity of prostein as a potential vaccine candidate for prostate cancer. To determine the presence of CD8 T,cells specific for naturally processed prostein-derived epitopes in healthy individuals, we developed and applied an in vitro stimulation protocol. Using this protocol, we identified CD8 T,cells specific for prostein in the peripheral blood of a male and a female donor. Prostein-specific CD8 T,cell clones specifically recognized prostein-expressing targets, including prostate tumor cell lines expressing the relevant HLA alleles. CD8 T,cell clones isolated from the male donor were significantly less effective in recognizing target cells compared to cells isolated from the female donor and appeared to recognize subdominant epitopes. The identification of a prostein-specificCD8 T,cell repertoire supports the development of prostein in vaccination strategies against prostate cancer. Furthermore, the naturally processed peptide epitopes identified provide tools for the development of peptide-based vaccination strategies against prostate cancer and for monitoring of prostein-specific responses in vaccinated patients. [source] Proteome-guided search for influenza A B-cell epitopesFEMS IMMUNOLOGY & MEDICAL MICROBIOLOGY, Issue 1 2009Guglielmo Lucchese Abstract The influenza A linear peptide epitopes recognized by murine antibodies, and currently cataloged at http://www.immuneepitope.org, were examined for the identity score to the host mouse proteome. It was found that almost all of the linear viral determinants are (or contain) regions formed by pentapeptide fragments with no or only very low similarity to the murine proteins. The present study adds to previous reports in suggesting a main role of amino acid sequence similarity in the modulation and definition of the B-cell epitope repertoire, inspiring innovative vaccine approaches able to avoid cross-reactive autoimmune collateral phenomena, and addressing future research in the study of immunity against the influenza A virus and infectious diseases in general. [source] Amoebic gill disease resistance is not related to the systemic antibody response of Atlantic salmon, Salmo salar L.JOURNAL OF FISH DISEASES, Issue 1 2010R S Taylor Abstract Amoebic gill disease (AGD) is a proliferative gill tissue response caused by Neoparamoeba perurans and is the main disease affecting Australian marine farmed Atlantic salmon. We have previously proposed that macroscopic gill health (,gill score') trajectories and challenge survival provide evidence of a change in the nature of resistance to AGD. In order to examine whether the apparent development of resistance was because of an adaptive response, serum was sequentially sampled from the same individuals over the first three rounds of natural AGD infection and from survivors of a subsequent non-intervention AGD survival challenge. The systemic immune reaction to ,wildtype'Neoparamoeba sp. was characterized by Western blot analysis and differentiated to putative carbohydrate or peptide epitopes by periodate oxidation reactions. The proportion of seropositive fish increased from 46% to 77% with each AGD round. Antibody response to carbohydrate epitope(s) was immunodominant, occurring in 43,64% of samples. Antibodies that bound peptide epitope were identified in 16% of the challenge survivors. A 1:50 (single-dilution) enzyme-linked immunosorbent assay confirmed a measurable immune titre in 13% of the survivors. There was no evidence that antibodies recognizing wildtype Neoparamoeba provided significant protection against AGD. [source] Epitope mapping of the neuronal growth inhibitor Nogo-A for the Nogo receptor and the cognate monoclonal antibody IN-1 by means of the SPOT techniqueJOURNAL OF MOLECULAR RECOGNITION, Issue 3 2007Hilke Zander Abstract Nogo-A is a potent inhibitor of axonal outgrowth in the central nervous system of adult mammals, where it is expressed as a membrane protein on oligodendrocytes and in myelin. Here we describe an attempt to identify linear peptide epitopes in its sequence that are responsible for the interaction either with the Nogo receptor (NgR) or with the neutralizing monoclonal antibody IN-1. Analysis of an array of immobilized overlapping 15,mer peptides covering the entire amino acid sequence of human Nogo-A (1192 residues) revealed a single epitope with prominent binding activity both towards the recombinant NgR and the IN-1 Fab fragment. Further truncation and substitution analysis yielded the minimal epitope sequence 'IKxLRRL' (x,,,P), which occurs within the so-called Nogo66 region (residues 1054,1120) of Nogo-A. The bacterially produced Nogo66 fragment exhibited binding activity both for the recombinant NgR and for the IN-1 Fab fragment on the Western blot as well as in ELISA. Unexpectedly, the synthetic epitope peptide and the recombinant Nogo66 showed cross-reactivity with the 8-18C5 Fab fragment, which is directed against myelin oligodendrocyte glycoprotein (MOG) as a structurally unrelated target. On the other hand, the recombinant N-terminal domain of Nogo-A (residues 334,966) was shown to specifically interact on the Western blot and in an ELISA with the IN-1 Fab fragment but not with the recombinant NgR, which is in agreement with previous results. Hence, our data suggest that there is a distinct binding site for the Nogo receptor in the Nogo66 region of Nogo-A, whereas its interaction with NgR is less specific than anticipated before. Although there probably exists a non-linear epitope for the neutralizing antibody IN-1 in the N-terminal region of Nogo-A, which is likely to be accessible from outside the cell, a previously postulated second binding site for NgR in this region (called Nogo-A-24) remains elusive. Copyright © 2007 John Wiley & Sons, Ltd. [source] Topology and patch-clamp analysis of the sodium channel in relationship to the anti-lipid a antibody in campylobacteriosisJOURNAL OF NEUROSCIENCE RESEARCH, Issue 15 2008Seigo Usuki Abstract An infecting strain VLA2/18 of Campylobacter jejuni was obtained from an individual with campylobacteriosis and used to prepare chicken sera by experimental infection to investigate the role of serum anti-ganglioside antibodies in Guillain-Barré syndrome. Both sera of the patient and chicken contained anti-ganglioside antibodies and anti-Lipid A (anti-Kdo2-Lipid A) antibodies directed against the lipid A portion of the bacterial lipooligosaccharide. The anti-Kdo2-Lipid A activities inhibited voltage-gated Na (Nav) channel of NSC-34 cells in culture. We hypothesized that anti-Kdo2-Lipid A antibody acts on the functional inhibition of Nav1.4. To test this possibility, a rabbit peptide antibody (anti-Nav1.4 pAb) against a 19-mer peptide (KELKDNHILNHVGLTDGPR) on the , subunit of Nav1.4 was produced. Anti-Nav1.4 pAb was cross-reactive to Kdo2-Lipid A. Anti-Kdo2-lipid A antibody activity in the chicken serum was tested for the Na+ current inhibition in NSC-34 cells in combination with ,-Conotoxin and tetrodotoxin. Contrary to our expectations, the anti-Kdo2-Lipid A antibody activity was extended to Nav channels other than Nav1.4. By overlapping structural analysis, it was found that there might be multiple peptide epitopes containing certain dipeptides showing a structural similarity with v-Lipid A. Thus, our study suggests the possibility that there are multiple epitopic peptides on the extracellular domains of Nav1.1 to 1.9, and some of them may represent target sites for anti-Kdo2-Lipid A antibody, to induce neurophysiological changes in GBS by disrupting the normal function of the Nav channels. © 2008 Wiley-Liss, Inc. [source] Viral peptide immunogens: current challenges and opportunitiesJOURNAL OF PEPTIDE SCIENCE, Issue 12 2007Ali Azizi Abstract Synthetic peptide vaccines have potential to control viral infections. Successful experimental models using this approach include the protection of mice against the lethal Sendai virus infection by MHC class I binding CTL peptide epitope. The main benefit of vaccination with peptide epitopes is the ability to minimize the amount and complexity of a well-defined antigen. An appropriate peptide immunogen would also decrease the chance of stimulating a response against self-antigens, thereby providing a safer vaccine by avoiding autoimmunity. In general, the peptide vaccine strategy needs to dissect the specificity of antigen processing, the presence of B-and T-cell epitopes and the MHC restriction of the T-cell responses. This article briefly reviews the implications in the design of peptide vaccines and discusses the various approaches that are applied to improve their immunogenicity. Copyright © 2007 European Peptide Society and John Wiley & Sons, Ltd. [source] Improved accuracy of cell surface shaving proteomics in Staphylococcus aureus using a false-positive controlPROTEINS: STRUCTURE, FUNCTION AND BIOINFORMATICS, Issue 10 2010Nestor Solis Abstract Proteolytic treatment of intact bacterial cells is an ideal means for identifying surface-exposed peptide epitopes and has potential for the discovery of novel vaccine targets. Cell stability during such treatment, however, may become compromised and result in the release of intracellular proteins that complicate the final analysis. Staphylococcus aureus is a major human pathogen, causing community and hospital-acquired infections, and is a serious healthcare concern due to the increasing prevalence of multiple antibiotic resistances amongst clinical isolates. We employed a cell surface "shaving" technique with either trypsin or proteinase- K combined with LC-MS/MS. Trypsin-derived data were controlled using a "false-positive" strategy where cells were incubated without protease, removed by centrifugation and the resulting supernatants digested. Peptides identified in this fraction most likely result from cell lysis and were removed from the trypsin-shaved data set. We identified 42 predicted S. aureus COL surface proteins from 260 surface-exposed peptides. Trypsin and proteinase- K digests were highly complementary with ten proteins identified by both, 16 specific to proteinase- K treatment, 13 specific to trypsin and three identified in the control. The use of a subtracted false-positive strategy improved enrichment of surface-exposed peptides in the trypsin data set to approximately 80% (124/155 peptides). Predominant surface proteins were those associated with methicillin resistance,surface protein SACOL0050 (pls) and penicillin-binding protein 2, (mecA), as well as bifunctional autolysin and the extracellular matrix-binding protein Ebh. The cell shaving strategy is a rapid method for identifying surface-exposed peptide epitopes that may be useful in the design of novel vaccines against S. aureus. [source] ORIGINAL ARTICLE: Presence of Antisperm Antibodies Reactive with Peptide Epitopes of FA-1 and YLP12 in Sera of Immunoinfertile WomenAMERICAN JOURNAL OF REPRODUCTIVE IMMUNOLOGY, Issue 6 2008Jessica Williams Problem Recent studies in several laboratories are focused on delineating sperm antigens that are relevant to fertility and examining involvement of antibodies to these antigens in human immunoinfertility. Our laboratory has characterized two such antigens, namely fertilization antigen (FA-1) and YLP12 dodecamer sequence that are involved in sperm-oocyte binding. The present study was conducted to examine the occurrence of isoantibodies to various peptide epitopes of human and murine FA-1 antigen and YLP12 peptide in sera of immunoinfertile and fertile women. Method of study Sera from 67 immunoinfertile and 19 fertile women were collected. Various peptides based up on human and murine FA-1 antigen and YLP12 were synthesized, and examined for immunoreactivity with these sera by using ELISA. Four immunodominant sequences, two each from human (hFA-182-97aa and hFA-1200-219aa) and mouse (mFA-12-19aa and mFA-1117-136aa) FA-1 antigen, were selected for the present study. Another human FA-1 sequence, hFA-1220-240aa, that was not in the immunodominant region was used as a control. Results For human FA-1 peptides, 41.8% of the immunoinfertile sera reacted positively (,2 SD units) with hFA-182-97aa, 24.6% (16/65) with hFA-1200-219aa, and 3% (2/66) with hFA-1220-240aa peptide. For two murine FA-1 peptides, 41.7% (25/60) of the immunoinfertile sera reacted positively with mFA-12-19aa, and 41.5% (27/65) with mFA-1117-136aa peptide. For the YLP12 dodecamer peptide, 43.3% (29/67) of the immunoinfertile sera reacted positively. None of the sera from fertile women reacted positively with any of these peptides. Conclusion In conclusion, our data indicate that the immunoinfertile women have circulating isoantibodies against at least two immunodominant peptide epitopes of human and murine FA-1 antigen and YLP12 peptide sequence. These peptides may find clinical application in the specific diagnosis and treatment of female infertility and contraceptive vaccine development. [source] Identification of immunodominant CD4+ T cell epitopes in patients with Yersinia -induced reactive arthritis by cytometric cytokine secretion assayARTHRITIS & RHEUMATISM, Issue 11 2006Andreas Thiel Objective In reactive arthritis (ReA), a bacteria-specific T cell response to the triggering microbe is detected in synovial fluid. So far, direct characterization of bacteria-specific T cells and identification of the immunodominant fine specificities remain difficult due to the lack of appropriate techniques. The aim of the present study was to directly determine the fine specificity of CD4+ T cells specific to ReA-associated bacteria-derived protein. Methods In 2 patients with Yersinia -induced ReA, live Yersinia Hsp60,specific CD4+ T cells were directly isolated from synovial fluid after stimulation with Yersinia -derived protein Hsp60 using a cytometric cytokine secretion assay. Generated short-term T cell lines were then tested in vitro for their peptide epitope specificity. Also, direct cross-reactivity of one line with Chlamydia - and human-derived Hsp60 was assessed. Results Generated short-term CD4+ T cell lines were highly antigen-specific and revealed single immunodominant peptide epitopes that were confirmed by direct testing with single peptides in both peripheral blood and synovial fluid cells. Yersinia Hsp60,specific T cells of one patient cross-reacted directly with human Hsp60. Conclusion Our results demonstrate the feasibility of direct assessment of live, potentially pathogenic, antigen-specific interferon-,+ CD4+ T cells taken from inflammatory lesions of patients with rheumatic diseases such as ReA. This might have implications not only regarding pathogenesis, but also in the design of new immunotherapies. [source] | |||||||||||||