B and T cell epitope mapping and study the humoral and cell mediated immune response to B-T constructs of YscF antigen of Yersinia pestis

YscF antigen, a type III secretion protein has recently been shown partial protection in murine model. Five peptides of YscF antigen were predicted using DNASTAR and T-cell prediction software. Peptides were synthesised and authenticated using competitive, direct binding immunoassay with anti YscF/peptide sera raised in mice. Peptide P1 and P2 were found to be B cell epitope while P3 was minor B cell epitope. P4 peptide was a pure T cell epitope based on lymphoproliferative response, cytokines profile and T-bet expression. Furthermore, with an intention to enhance immunogenicity, three B-T constructs were designed between the above epitopes. Conjugate B1T1 and B2T1 showed higher serum IgG/IgA titre, respectively, as well as high secretory IgA plus secretory component (Sc) both in lung and intestinal washes. Also, these conjugates showed high T-cell proliferation in addition to higher Th1 type cytokines (IFN-γ and IL-2) in cells obtained from spleen, lamina propria and Peyer's patches. B3T1 stimulated cells showed moderate levels of IFN-γ and IL-2 but higher levels of IL-4. This study demonstrates superior immunogen of B1T1 and B2T1 of YscF antigen to be exploited as vaccine candidate for plague.