The Research Of Recombinant IFN-a With PPV VP2-JEV E DNA Vaccine's Immunogenicity

Clinically,sow reproductive disorders caused by infection of Parvoviruses or Japanese encephalitis virus or mixed infection of them can show such symptoms as miscarriage, stillbirth, mummified fetus, abnormal fetus, weak fetus and the boar infertility and so on,caused great damage to the sustainability development of pig industry. Parvovirus VP2 protein and the Japanese encephalitis virus E protein as the virus particles major structural protein was confirmed to had good immunogenicity,and could induced immune response with producing corresponding antibodies. As an important cell activity factor in the body, IFN-a played an important role in many ways such as anti-virus, anti-tumor activity of cell proliferation and immune activity regulation.In this experiment, the PCR amplified poIFN-a gene was clonged into pMD19-T Simple vector,and the recombinant plasmid pMD-IFNa was obtained. Then,the IFN-a gene was into the eukaryotic expression vectors PcDNA3.1-E-VP2,which had been integrated with the major epitope of PPV VP2 and JEV E gene,by the mean of double enzyme digestion with restriction endonuclease enzyme Nheâ… and Kpnâ… .The recombinant plasmid was named PcDNA3.1-IFNa-E-VP2.Then healthy experimental mouse were immunized using the recombinant plasmid DNA and withPcDNA3.1-E-VP2+pMD-IFNa, PcDNA3.1-E-VP2and 0.9% NaCl as control. The dynamic variation of mice blood T lymphocytes and the antibody variation of PPV and JEV in serum were measured.The results confirmed that,the ratio of CD3+, CD4+ T lymphocyte and peripheral blood lymphocyte of immunized mouse raised in a certain degree, CD8+T lymphocyte fell in the 7-14d after immunization, and then raised. Relatively high level of PPV, JEV specific antibody was detected from serum at the same time, and this indicated that recombinant plasmid DNA with IFNa gene had good immunogenicity. IFN-a had a certain role in enhancement and supporting of immune action. The recombinant plasmid DNA have the potential to be constructed into new type PPV-JEV bivalent genetic engineering vaccine.