Screening And Identification Of H-2d Restricted T Cell Epitope Of Structural Protein VP1 Of Foot-and-mouth Disease Virus Type A Strain AF/72

Epitope vaccine, as a new kind of vaccine, has become one of research hot spots of FMD vaccine. Screening and identification of T cell epitope of FMDV is significant to developing high-efficiency vaccines and further research due to cellular immunity play an important role in immune response of FMDV infection and structural protein VP1 include main antigen sites of FMDV.In this study, The Complete sequence of VP1 of FMDV AF / 72 strains was obtain through cloning and sequencing, and then VP1 protein have high-level expressed in E. coli using the method of codon optimization.The detection of SDS-PAGE and Western Blotting suggest that the molecular weight of expressed VP1 proteins correspond to expected and keep a good immunogenicity. Meanwhile,30 the H-2d Restricted T Cell Epitope of VP1 have been forecast and synthesis using the relative bioinformatics software.The VP1 protein was used for immunizing BALB/c mice and spleen lymphocytes were isolated, and vitro common training stimulus was carried out to potential H-2Dd, H-2Kd and H-2Ld restricted T cell epitope.The H-2Kd restricted T cell epitope pK1 (AYHKGPFTRL) and H-2Dd restricted T cell epitope pD7 (GFIMDRFVKI) were identified through lymphocyte proliferation assay and IFN-γELISPOT experiments.This study provides method and basis through screen and identifies H-2d restricted T cell epitope of structural protein VP1 of foot-and-mouth disease virus type A. in addition, screening and identification of mouse's CTL epitope contribute to understanding the effect of cytotoxic lymphocytes against FMDV.