Identification of immunodominant epitope of F1 antigen of Yersinia pestis.
The F1 antigen of Yersinia pestis has been identified as one of the major protective antigens of this bacterium. The present study aims to delineate major and minor antigenic sites of F1 antigen. Using algorithmic predictions, five peptide sequences (P1, P2, P3, P4 and P5) spanning the C-terminal region were identified and synthesized. Antibodies were generated in mice against the peptides, native F1 protein and polymerized F1 antigen using liposomes as mode of immunization. Cross-reactivity between F1 antigen and peptides was tested using both solid and solution phase assays. Similar assays were done with rabbit anti-F1 sera. Competitive inhibition assays using a different combination of antisera and competing antigen identified P2 peptide FFVRSIGSKGGKLAAGKYTDAVTV (142-165) as the immunodominant sequence. The results indicate that this sequence appears to be exposed on the surface of F1 molecule. In a solid phase binding assay, P2 peptide was recognized even at high F1 antisera dilution. However, when antisera raised to different peptides were tested for binding to F1 antigen, antisera to P4 peptide showed maximal immunoreactivity. This implies more accessibility of this region during immobilization on solid surface. There was consistency in the results obtained for different strains of mice as well as for the rabbit antisera. Such a sequence of F1 antigen, which is recognized widely in animals of different genetic background, would be useful for diagnosis and subunit vaccine.
Sabhnani
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Department of Biochemistry, All India Institute of Medical Sciences, Ansari Nagar, New Delhi, India.
Rao
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DN
D000595:Amino Acid Sequence; D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry* / Q000276:immunology*; D001667:Binding, Competitive; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D016056:Immunodominant Epitopes; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D008969:Molecular Sequence Data; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D015010:Yersinia pestis / Q000276:immunology*
D000907:Antibodies, Bacterial; D001426:Bacterial Proteins; D016056:Immunodominant Epitopes; D010455:Peptides; C086725:caf1 protein, Yersinia pestis