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1.
IpaB, invasion plasmid antigen B, of Shigella flexneri is a 62-kDa protein required for invasion of intestinal epithelial cells. IpaB is also one of several major protein antigens recognized by the humoral immune systems of most humans and monkeys after infection with shigellae. Computer analysis of the deduced IpaB amino acid sequence indicates that an alpha-helical structure is likely through much of the molecule. Homology searches with protein data banks show that one alpha-helical domain between amino acid residues 95 and 181 has a moderate level of identity with myosin and streptococcal M protein. By using a monoclonal antibody (2F1) which recognizes an epitope in the amino-terminal third of the IpaB protein, it was possible to demonstrate a cross-reactive epitope(s) on skeletal muscle myosin. Epitope mapping localized the 2F1 epitope to three noncontiguous regions of the IpaB protein within the alpha-helical domain that contains homology with myosin. Antibodies produced in rabbits immunized with synthetic peptides from one of the 2F1 epitope regions (residues 99 to 110) of IpaB were capable of reacting with IpaB as well as myosin. Furthermore, sera from several monkeys previously infected with S. flexneri 2a contained antibodies to IpaB pep 101-116 (IpaB peptide 101-116) and also myosin. Sera from animals with antibodies against other IpaB peptides did not contain antibodies against myosin. 相似文献
2.
Human monoclonal antibodies against a recombinant HIV envelope antigen produced by primary in vitro immunization. Characterization and epitope mapping. 总被引:4,自引:0,他引:4
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M Ohlin P A Broliden L Danielsson B Wahren J Rosen M Jondal C A Borrebaeck 《Immunology》1989,68(3):325-331
Peripheral blood lymphocytes from healthy, HIV sero-negative blood donors have been in vitro immunized using penv9, a recombinant fragment of the envelope of HIV-1. This primary in vitro immunization followed by Epstein-Barr virus (EBV) transformation and somatic cell fusion subsequently gave rise to several specific anti-penv9 monoclonal antibodies (MO28, MO30 and MO43) of mu isotype. The hybridomas have been kept in culture for over 6 months and the antibody productivity for MO30 was measured to 18 micrograms x (24 hr x 10(6) cells)-1. The fine specificity of the antibodies was mapped by a peptide inhibition enzyme immunoassay, using overlapping synthetic pentadeca peptides covering the whole penv9. These human monoclonal antibodies exhibited a similar epitope specificity directed against a non-sequential determinant, including the amino acids 632-646, 677-681 and 687-691. This specificity is very rarely found in immune sera from seropositive patients and presently not reported in human monoclonal antibodies derived from in vivo immunized individuals, indicating that different antibody specificities can be obtained by the in vitro immunization technology. These human monoclonal antibodies did not neutralize HIV. The results presented here demonstrate the feasability of generating human monoclonal antibodies against HIV by primary in vitro immunizations, thereby avoiding the use of lymphocytes derived from infected patients when human monoclonal antibodies for therapeutic purposes are to be produced. 相似文献
3.
Primate antibody response to immunotoxin: serological and computer-aided analysis of epitopes on a truncated form of Pseudomonas exotoxin.
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NLysPE38 is a 38-kDa derivative of Pseudomonas exotoxin (PE) in which domain Ia (amino acids 1 to 252) and part of domain Ib (365 to 380) are deleted and an 11-amino-acid N-terminal peptide is added. LMB-1 is an immunotoxin in which the monoclonal antibody B3 is coupled to NLysPE38 near its N terminus. LMB-7 is a single-chain immunotoxin in which the Fv fragment of B3 is fused to PE38. To identify the antigenic regions of PE38, 12 polyclonal serum samples from monkeys immunized with the immunotoxins LMB-1 (six monkeys) and LMB-7 (six monkeys) were tested for their reactivity to a panel of 120 synthetic, overlapping peptides representing the amino acid sequence of NLysPE38. The antibody responses to peptides were similar among the 12 serum specimens, identifying several major immunodominant B-cell epitopes. Predominant reactivity was seen in six locations: amino acids 272 to 287, 341 to 359, 504 to 516, 540 to 564, and 573 to 591 and the C-terminal amino acids 591 to 613. The sera did not react with approximately 75% of the peptides. Furthermore, a computer-aided analysis was done to predict the immunologically relevant areas and revealed the same antigenic regions defined by serum reactivity to peptides. Competition enzyme-linked immunosorbent assays and neutralization assays were performed with domain II, III, or III plus Ib of PE38 and confirmed the immunodominance of domain III. To analyze the role of specific amino acids in antibody binding, individual amino acids of PE38 with large accessible surface areas were altered by site-directed mutagenesis. These results also show that the predicted areas of immunogenicity agree with the reactivity of the anti-PE38 antibodies to peptides and to the mutants of PE. 相似文献
4.
Identification of a nucleocapsid protein as a specific serological marker of human herpesvirus 6 infection. 总被引:8,自引:1,他引:8
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M Yamamoto J B Black J A Stewart C Lopez P E Pellett 《Journal of clinical microbiology》1990,28(9):1957-1962
Enveloped whole virions and nucleocapsids of human herpesvirus 6 (HHV-6) strain Z29 were purified from supernatant fluids of infected human cord blood lymphocytes by filtration through polyvinylpyrrolidone-treated filters, banding on a Nycondenz step gradient, and centrifugation through two successive continuous sucrose gradients. More than 20 proteins ranging in molecular weight from less than 30,000 to more than 200,000 were identified in preparations of purified whole virions labeled with [35S]methionine and [35S]cysteine. Immunogenic virion proteins of HHV-6 were identified in immunoblot assays with human immune sera, immune sera generated from mice immunized with purified whole virions or purified nucleocapsids, and a monoclonal antibody generated from a mouse immunized with purified nucleocapsids. The sera and the monoclonal antibody reacted strongly with a 101-kilodalton protein in the immunoblots, suggesting that the protein is a component of the nucleocapsid. Human sera lacking HHV-6-specific antibodies and seropositive for one or more of the other human herpesviruses failed to react with this protein, indicating that it is a specific serologic marker for HHV-6 infection. 相似文献
5.
Y. Tamura J. Kawaguchi† N. Serizawa† K. Hirahara‡ A. Shiraishi‡ H. Nigi§ Y. Taniguchi¶ M. Toda S. Inouye T. Takemori M. Sakaguchi 《Clinical and experimental allergy》2003,33(2):211-217
BACKGROUND: Japanese cedar (Cryptomeria japonica; CJ) pollinosis has been reported to occur naturally in Japanese monkeys (Macaca fuscata) as well as in humans. Most human patients and monkeys with pollinosis have specific IgE for Cry j 2, a major allergen of CJ pollen. OBJECTIVE: The main purpose of this study was to identify IgE B cell epitopes of Cry j 2 using a synthetic peptide in humans, monkeys and mice. METHODS: We synthesized 38 overlapping peptides that span the entire length of Cry j 2. We examined the B cell epitopes of Cry j 2 that are recognized by IgE in the sera of human patients and monkeys with pollinosis and immunized mice using synthetic peptides of Cry j 2. We also examined the reaction of Cry j 2-specific mouse monoclonal IgG antibodies to the peptides. Furthermore, we conducted a histamine release assay with leucocytes from a pollinosis patient using human serum albumin (HSA) conjugated with the peptides as a B cell epitope. RESULTS: We found that 16 of the 20 pollinosis patients who had specific IgE to Cry j 2 also exhibited IgE reaction with some Cry j 2 peptides. Of these 16 patients, 10 exhibited IgE reaction with Cry j 2 peptide no. 13 (121GQCKWVNGREICNDRDRPTA140). Five of the seven monkeys with CJ pollinosis exhibited a reaction with peptide no. 13. Furthermore, IgE in mice immunized with Cry j 2 and two mouse monoclonal IgG antibodies reacted with peptide no. 13. Peptide no. 13-conjugated HSA showed the release of histamine from basophils. Furthermore, to determine the minimum epitope in peptide no. 13, we conducted an enzyme-linked immunosorbent assay inhibition test. The core of the epitope in humans, monkeys and mice was 124KWVNGREI131. CONCLUSION: We found that 124KWVNGREI131 is an important B cell epitope recognized by IgE in humans, monkeys and mice. 相似文献
6.
Stadler BM Zürcher AW Miescher S Kricek F Vogel M 《International archives of allergy and immunology》1999,118(2-4):119-121
We have defined epitopes on human IgE by screening different phage display random peptide libraries with a monoclonal anti-IgE antibody termed BSW17. The selected mimotopes and epitopes within the Cepsilon3 and Cepsilon4 region of IgE induced antibodies that were nonanaphylactogenic and had biological activity similar to BSW17. The chemically synthesized and KLH-coupled IgE epitopes or mimotopes were used to induce an anti-IgE response in rhesus monkeys. The immunized rhesus monkeys were subsequently protected in a PCA test when sensitized with human IgE and triggered with the corresponding allergen. Furthermore, using the same monoclonal anti-IgE antibody, we also generated an anti-idiotypic antibody that showed sequence homology with the IgE epitope in the Cepsilon3 domain. This anti-idiotypic antibody as well as the mimotopes were then used in a mouse model to induce orally an anti-IgE immune response. For this purpose mice were fed by intragastric gavages with bacteriophages displaying the small IgE-homologous structures. Orally immunized mice produced serum anti-IgE antibodies that were inhibited by BSW17 suggesting that it may be possible to induce a systemic anti-IgE response orally. 相似文献
7.
Human monoclonal antibody fragments neutralizing wild-type and vaccine strains of yellow fever (YF) virus (genotypes West Africa I + II, East/Central Africa, 17D-204-WHO) were generated by repertoire cloning from YF patients. Analysis of virus escape variants identified amino acid (aa) 71 in domain II of the envelope glycoprotein (E) as the most critical residue for neutralization, with aa 153-155 in domain I contributing to the epitope. These data confirm the previous mapping of YFV neutralizing epitopes using mouse monoclonal antibodies but suggest that a conformational epitope could be formed by amino acids from domains I and II opposing each other in the dimeric form of the E protein. While the sera of the YF patients showed up to 10-fold reduced neutralizing activity against the 17D escape variants, sera from 17D vaccinees retained their neutralizing titers. Mutations in this major neutralizing epitope of YFV thus do not seem to carry the risk of immune escape in persons immunized with the YFV-17D vaccine. 相似文献
8.
Identification of antibody epitopes in the CB-11 peptide of bovine type II collagen recognized by sera from arthritis-susceptible and -resistant rhesus monkeys.
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S Turner N P Bakker B A t Hart P J Holt K Morgan 《Clinical and experimental immunology》1994,96(2):275-280
Sera from eight rhesus monkeys that had been immunized with native bovine type II collagen were tested for antibodies to cyanogen bromide peptides (CB peptides) of type II collagen by Western blotting. The monkeys produced IgG antibodies to a number of different CB peptides, with five out of eight animals producing antibodies to the CB-11 peptide (four arthritic, one non-arthritic). Antibody epitopes on the CB-11 peptide of bovine type II collagen recognized by these sera were investigated by epitope mapping. Peptides (8-mers overlapping by seven amino acids) representing the CB-11 region were synthesised and the sera screened for binding to these peptides to determine areas of high IgG antibody binding to this region of type II collagen. The profiles obtained were not identical, though there were some epitopes that were commonly recognized. Antibodies to one epitope, also present in human type II collagen, were found only in the sera of two animals with the severest arthritis. The technique of epitope mapping has successfully identified a number of epitopes within the CB-11 peptide of type II collagen recognized by antibodies from bovine type II collagen-immunized monkeys. Studies on the relevance of responses to the identified epitopes can now be undertaken. 相似文献
9.
Minoru Ayata Tohru Sugano Tsugiya Murayama Daitoku Sakamuro Tsutomu Takegami Yoh-Ichi Matsumoto Toru Furukawa 《Journal of medical virology》1994,43(4):386-392
The amino-terminal portion of human cytomeg-alovirus glycoprotein B (HCMV-gB) was expressed as a fusion protein to analyze the neutralizing epitope recognized by human monoclonal antibody C23 and the humoral immune response to this epitope. The linear neutralizing epitope was further localized to the pep-tide within 17 amino acids (position 68-84) which were conserved between two HCMV laboratory strains. Ten out of 17 HCMV-seropositive human sera contained the antibody against this epitope. Although seven sera were negative for reacting with the fusion protein, the viruses isolated from the same patients retained the epitope. The immunogenicity of the epitope and the possible application of C23 human monoclonal antibody for passive immunization against HCMV infections are discussed. © 1994 Wiley-Liss, Inc. 相似文献
10.
Use of recombinant epitopes to study the heterogeneous nature of the autoantibodies against thyroid peroxidase in autoimmune thyroid disease. 总被引:1,自引:0,他引:1
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Microsomal antigen is often recognized by the sera from patients with autoimmune thyroid disease (AITD). Human thyroid peroxidase (hTPO) is the main component of this antigen. In a previous study, we expressed hTPO cDNA as fusion proteins in prokaryotic vector; we thereby defined seven antigenic peptides by using two rabbit polyclonal anti-hTPO antibodies. In the present study we used the seven epitopes and three widened peptides to define the reactivity pattern of 61 sera from patients with AITD. Thirty-eight of them reacted against at least one of the seven hTPO-restricted epitopes; 14 were negative against the seven determinants but recognized one or two of the extended peptides. Thus, the antibody response against hTPO appeared to be highly heterogeneous in AITD patient sera. Moreover, we demonstrated that the immunodetection of the hTPO on Western blotting with deoxycholate solubilized microsomes can be perfectly correlated with the recognition of one of the epitopes in the region 554-735. 相似文献
11.
Lactobacilli are nonpathogenic gram-positive inhabitants of the normal human intestine known for their health-promoting effects. In our earlier work, it is shown that human monoclonal antibody isolated from sera of a patient with Waldenstrom macroglobulinaemia possess innate antibody characteristics and binds to lactic acid bacteria. According to the immune network model, immunization with this bacteria could induce the perturbations in immune system that might result in production of anti-Lactobacillus antibodies, human monoclonal antibody like (Ab1) and anti-idiotypic antibody (Ab2). In this study, BALB/ c mice were immunized with two doses of bacteria Lactobacillus acidophilus in complete and incomplete Freund's adjuvant and phosphate-buffered saline (PBS), respectively. Seven days after the last immunization, sera from immunized mice were collected and the presence of Lactobacillus-specific Ab1 and Ab2 were determined by ELISAs. In the sera of immunized mice, antibodies specific to bacteria Lactobacillus acidophilus were shown. The concentration of Lactobacillus-specific antibodies was higher in the sera of hyperimmunized mice (mice immunized with 1 mg of IgM DJ) than in sera of mice immunized with 100 times lower doses of immunogen (0.01 mg per doses). Moreover, Ab1 and Ab2 antibodies were detected in the sera of Lactobacillus-hyperimmunized mice. In this study, we have shown the idiotypic network interactions in mice immunized with bacteria Lactobacillus acidophilus . 相似文献
12.
A monoclonal competitive radioimmunoassay (CompRIAm) which detects antibody to herpesvirus simiae (B virus) in monkey and human sera and antibody to SA8 virus in monkey sera but not antibody to herpes simplex virus in human sera is described. Of 232 serum samples from wild-caught cynomolgus monkeys, 117 serum samples were positive when tested by CompRIAm. The results were in close agreement (97.5%) with B virus neutralizing antibody results on the same sera. Sera from 97 wild-caught rhesus monkeys and 92 wild-caught baboons were also tested. The CompRIAm was able to differentiate between sera that had neutralizing antibody to B virus and SA8 virus and those that did not, although the discrimination was not as clear as that in the tests on cynomolgus monkey sera. Sequential sera from two humans with confirmed cases of B virus infection were tested by CompRIAm. B virus antibody was detected in sera from both humans. None of 237 other serum samples from blood donors and patients attending sexually transmitted disease clinics reacted in the CompRIAm. 相似文献
13.
Human humoral responses to antigens of Mycobacterium tuberculosis: immunodominance of high-molecular-mass antigens. 总被引:4,自引:0,他引:4
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S Laal K M Samanich M G Sonnenberg S Zolla-Pazner J M Phadtare J T Belisle 《Clinical and Vaccine Immunology : CVI》1997,4(1):49-56
The selection of antigens of Mycobacterium tuberculosis for most studies of humoral responses in tuberculosis patients has been restricted to molecules that were either immunodominant in immunized animals or amenable to biochemical purification rather than those that were reactive with the human immune system. Delineation of antigens that elicit humoral responses during the natural course of disease progression in humans has been hindered by the presence of cross-reactive antibodies to conserved regions on ubiquitous prokaryotic antigens in sera from healthy individuals and tuberculosis patients. The levels of cross-reactive antibodies in the sera were reduced by preadsorption with Escherichia coli lysates, prior to studying their reactivity against a large panel of M. tuberculosis antigens to which the human immune system may be exposed during natural infection and disease. Thus, reactivity against pools of secreted, cellular, and cell wall-associated antigens of M. tuberculosis was assessed by an enzyme-linked immunosorbent assay (ELISA). Initial results suggested that the secreted protein preparation contained antigens most frequently recognized by the humoral responses of pulmonary tuberculosis patients. The culture filtrate proteins were subsequently size fractionated by preparative polyacrylamide gel electrophoresis, characterized by reaction with murine monoclonal antibodies to known antigens of M. tuberculosis by an ELISA, and assessed for reactivity with tuberculous and nontuberculous sera. Results show that a secreted antigen of 88 kDa elicits a strong antibody response in a high percentage of patients with pulmonary tuberculosis. This and other antigens identified on the basis of their reactivity with patient sera may prove useful for developing serodiagnosis for tuberculosis. 相似文献
14.
B. Evengård† I. WOLOWCZUK‡§ M. MARGUERITE‡ L. HAMMARSTRÖM E. SMITH C. AURIAULT‡ 《Scandinavian journal of immunology》1994,40(6):618-622
Consecutive sera from before and after treatment were collected from 23 patients with a Schistosoma mansoni infection, seven of whom had an early infection. All sera were analysed for IgGl, IgG3 and IgG4 antibody activity against three peptides from the protein Sm 28 GST (24–43, 115–131 and 140–153 aa). In addition, sera from 14 patients, four with an early infection and 10 patients with a chronic infection, were analysed for IgG and IgA antibody activity using seven peptides derived from the protein Sm 28 GST. This molecule has previously demonstrated protective activity against infection in various experimental models.
The results are indicative of a subclass-related epitope specificity of the antibodies. Moreover, reactivity to one of the peptides (158–175 aa) was significantly associated with a chronic infectious status. 相似文献
The results are indicative of a subclass-related epitope specificity of the antibodies. Moreover, reactivity to one of the peptides (158–175 aa) was significantly associated with a chronic infectious status. 相似文献
15.
Serum antibodies to Porphyromonas gingivalis block the prostaglandin E2 response to lipopolysaccharide by mononuclear cells. 总被引:1,自引:0,他引:1
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The ability of rabbit and monkey immune sera to neutralize prostaglandin E2 (PGE2) production by human monocytes stimulated with lipopolysaccharide (LPS) was examined. CD14-dependent LPS activation of PGE2 was examined under assay conditions which allowed the comparison of preimmune and immune sera. Serum obtained from rabbits immunized with formalin-fixed Porphyromonas gingivalis cells dramatically reduced the amount of PGE2 produced in response to LPS obtained from three different strains of P. gingivalis but not that from Escherichia coli or Bacteroides fragilis. In addition, a significant reduction in the mean PGE2 level was observed in the presence of sera from immunized but not control monkeys employed in a vaccine trial. Immune serum samples from five of nine immunized monkeys were able to reduce LPS-induced production of PGE2 by greater than 50% compared to that in the corresponding preimmune sera. Immune monkey serum, similar to immune rabbit serum, blocked PGE2 production in response to P. gingivalis LPS but not E. coli LPS. These data demonstrate that immunization with P. gingivalis whole cells can elicit an antibody response that is able to block the PGE2 response to LPS. Neutralization of LPS-mediated inflammatory mediator production may account in part for the observed suppression of alveolar bone loss in immunized monkeys. 相似文献
16.
Production and characterization of mouse monoclonal antibodies to allergenic epitopes on LolpI (Rye I). 总被引:1,自引:0,他引:1
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This study describes the production and characterization of mouse monoclonal antibodies (Mab) to LolpI (Rye I), the major allergen of rye-grass pollen. Hybridoma cell lines were screened for the secretion of Mab, the binding of which to labelled LolpI could be inhibited by pooled human sera containing IgE and IgG anti-LolpI antibodies. Twenty monoclonal hybridoma cell lines were expanded, and their product was purified and characterized. Each Mab bound to LolpI but not to unrelated antigens. Cross-inhibition studies allowed the grouping of these 20 Mab into five families differing by their epitope specificity. One representative Mab of each family was tested for its ability to inhibit human IgE/IgG anti-Lolp-I from 15 allergic donors as well as for its inhibition by affinity-purified anti-LolpI antibodies isolated from 14 donors. The results indicated that each family of Mab was specific for an epitope that was identical or close to that recognized by some human anti-LolpI antibodies. It is further shown that human IgE/IgG anti-LolpI are heterogeneous with some patients reacting preferentially with one or two epitopes defined by the Mab LolpI, and others reacting with epitopes different from those identified by the Mab LolpI. 相似文献
17.
Mapping of antigenic sites on the nucleocapsid protein of the severe acute respiratory syndrome coronavirus 总被引:8,自引:0,他引:8
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Antigenic sites on the nucleocapsid (N) protein of severe acute respiratory syndrome (SARS) coronavirus (SARS-CoV) were mapped by Pepscan analysis with overlapping peptides that span the N protein sequence. Two major immunodominant epitopes located in the C-terminal region (amino acids [aa] 362 to 412) and middle region (aa 153 to 178) reacted with more than 75% of sera from SARS patients. Several minor immunodominant epitopes were reactive with about 50% of the SARS sera. Antisera from mice immunized with inactivated SARS-CoV recognized the two major immunodominant epitopes and one antigenic site located adjacent to the N-terminal region (aa 76 to 101), which did not react with the sera from SARS patients. Several monoclonal antibodies against SARS-CoV bound to the N- or C-terminal antigenic sites. These results suggest that the above antigenic sites on the N protein are important in eliciting humoral immune response against SARS-CoV in humans and animals and can be used as antigens for developing diagnostic tests. 相似文献
18.
P von Landenberg C von Landenberg M Grundl G Schmitz J Sch?lmerich I Melchers 《Journal of autoimmunity》1999,13(1):83-87
Human B cell hybridomas were established to define new autoantigens of importance for autoimmune diseases such as rheumatoid arthritis (RA). One lgG1, lambda monoclonal antibody (FKN-E12), was derived from synovial B lymphocytes of a patient with sero-negative RA. The purified lg was used to select specifically binding peptides from a random peptide phage display library. Only one epitope with the heptamer sequence HLTFGPG was detected and named RASFp1. Very similar and partly identical sequences are found in the variable region of lg kappa light chains in position 96-101, at the junction of framework 2 and the J-region. The antibody FKN-E12 was shown to detect the epitope RASFp1 also on human lgG kappa chains, but only in a specific conformation. The aim of the present study was to analyse human sera from patients with autoimmune diseases, non-autoimmune inflammatory diseases and healthy blood donors for the presence of lgG binding to RASFp1. For this purpose a 15-mer-peptide was synthesized containing RASFp1 within Vk-derived flanking regions, and an ELISA assay established. Sera of 142 individuals were studied. Only <5% of the control sera including sera from patients with non-autoimmune inflammations were positive. In contrast, 45% of sera from patients with RA or SLE contained RASFp1-binding antibodies. Within the 40 RA sera analysed so far, rheumatoid factors and RASFp1-binding antibodies have shown no correlation with each other. 相似文献
19.
C. P. MULLER T. SCHROEDER R. TU N. H C. BRONS G. JUNG F. SCHNEIDER K. H. WIESMÜLLER 《Scandinavian journal of immunology》1993,38(5):463-471
Infection or immunization with measles virus induces a protective immune reaction including neutralizing antibodies against the haemagglutinin and fusion protein. The reactivity of the polyclonal IgG response of sera obtained from late convalescent donors was studied, using overlapping 15mer peptides covering the complete sequence of the measles virus haemagglutinin. Most sera reacted with a similar set of peptides generating a characteristic binding pattern. The reactive peptides correspond to a region mediating cell hemolysis (aa310–325), to regions which serve as targets to neutralizing antibodies and to a putative transmembrane region (aa35–58). The latter region contains also a human T-cell epitope providing evidence of a non-random association of T- and B-cell epitopes. We also immunized different strains of mice and rabbits with measles virus. In contrast to the human sera, animal sera with strong neutralizing activities did not react with any of the H-protein peptides. The mostly weak reactivities with the linear sequences contrast with the strong neutralizing activities of the human or animal antibodies, suggesting that these primarily recognize the fusion protein or conformational epitopes of the haemagglutinin protein. 相似文献
20.
Jensen Gasim Ismail Gaafar Kurtzhals Kemp El Hassan Kharazmi & Theander 《Scandinavian journal of immunology》1998,48(1):103-109
Native kinetoplastid membrane protein-11 (KMP-11), purified from crude extracts of Leishmania donovani parasites, activates T cells from individuals who have recovered from visceral leishmaniasis. In this work we used three 38-mer peptides spanning the amino acid sequence of the L. donovani KMP-11 as solid-phase ligands in enzyme-linked immunosorbent assays (ELISAs) and as stimulating antigens in lymphoproliferative assays in order to evaluate humoral and cellular immune responses to well-defined sequences of the protein. Antibody reactivity against the three peptides was measured in plasma from 63 Sudanese visceral leishmaniasis patients (VL) and the percentage of patients with anti-KMP-11 antibodies in ELISA were 37% (KMP-11-1), 30% (KMP-11-2) and 58% (KMP-11-3). The fraction of VL patients with measurable antibody reactivity in one or more of the three ELISAs was 79%. Cross-reactivity to the KMP-11 peptides was detected in plasma from Sudanese patients suffering from Leishmania major infections and in plasma from Sudanese and Danish patients infected with Plasmodium falciparum . In lymphoproliferative assays, 10 of 17 PBMC isolates from donors previously infected with L. donovani showed a response to one or more of the three KMP-11 peptides. 相似文献