非洲猪瘟病毒E248R蛋白的表达和免疫原性评价OA

To prepare recombinant African swine fever virus(ASFV)E248R protein using the baculovirus expression system and evaluate its immunogenicity,an E248R recombinant baculovirus expression vector was constructed.The target protein was expressed and purified in Spodoptera frugiperda Sf9 cells,and its ability to induce humoral and cellular immune responses was evaluated in mice.The results showed that the recombinant ASFV E248R protein was efficiently expressed in Sf9 cells and specifically recognized by ASFV-positive serum.Animal immunization experiments demonstrated that the recombinant ASFV E248R protein induced high levels of specific immunoglobulin G(IgG)antibodies in mice.The cell proliferation assay(CCK-8)showed that the stimulation index(SI)of splenic lymphocytes in the ASFV E248R recombinant protein-immunized group was significantly higher than that in the PBS control group(P<0.000 1).Enzyme-linked immunospot assay(ELISpot)revealed that the recombinant ASFV E248R protein significantly promoted interferon-γ(IFN-γ)secretion by lymphocytes(P<0.001).Flow cytometry analysis showed that the recombinant ASFV E248R protein significantly increased the proportions of CD4⁺(P<0.000 1)and CD8⁺ T lymphocytes(P<0.01).These results indicate that the ASFV E248R recombinant protein was successfully prepared and could induce strong humoral and cellular immune responses in mice,suggesting its potential as a candidate antigen for ASFV subunit vaccines.