The Construction And Immunogenicity Studies Of Recombinant Rabies Virus-based Vectored PPRV Vaccine

Peste des petits ruminants（PPR）is one of the most contagious and fatal animal diseases in the world.PPR is classified as a category A disease by the Office international des épizooties（OIE）and is included within the diseases and infections of goats and sheep of Terrestrial Animal Health Code.PPR is also widespread in wild ruminants and can spread across international borders.Its clinical and pathological symptoms are similar to rinderpest and human measles.The high morbidity and mortality of PPR will cause devastating economic losses to the global livestock industry.The presence of PPR,especially in developing countries in Africa,Asia and Europe,will continue to undermine the food security,livelihoods and trade of pastoralist,and become a threat to biodiversity and ecosystem health.Currently,PPR is the target disease of the global eradication campaign led by the OIE and the Food and Agriculture Organization of the United Nations（FAO）.In this experiment,the rabies virus reverse genetic system was used to rescue the recombinant rabies virus expressing the envelope glycoprotein H（hemagglutinin protein）or F（fusion protein）gene of Peste des petits ruminants virus（PPRV）.To develop a dual inactivated vaccine that can prevent PPR and rabies,and provide an effective bivalent inactivated vaccine candidate for the eradication of PPR and the prevention of rabies in livestock:（1）The H and F genes of the PPRV China/Tibet/Geg/07-30 strain were obtained by chemical synthesis,and were respectively connected to the rabies virus vectorSRV9 through the Bsi WⅠ and PmeⅠ restriction sites.Two full-length infectious clones p D-SRV9-PM-PPRV-H and p DSRV9-PM-PPRV-F expressing PPRV H and PPRV F genes were constructed respectively.After identification by enzyme digestion and sequencing,BSR/T7 cells were transfected with the helper plasmids,and the two recombinant viruses rSRV9-H and rSRV9-F were rescued.They were identified at the gene and protein levels,and the correct viruses were passaged and stored.（2）Two recombinant viruses were cultured in large quantities,inactivated and purified,mixed with adjuvant and used as an immunogen to immunize BALB/c mice,each recombinant virus was immunized at100μg/mouse,and set up a control group.Blood was collected regularly,and the spleen of the mice was collected on the 35 th day.The serum and spleen cells were tested for immune indicators.The immunogenicity of the two vaccines was analyzed.It was found that the two vaccines could produce specific Ig G antibodies,and produced PPRV and RABV neutralizing antibodies.,and can induce Th1 and Th2 immune responses,and is expected to become a bivalent vaccine that can simultaneously prevent PPRV and RABV infection,and the immune effect of rSRV9-H is better than rSRV9-F.