Research On Expression Of RABV Protein Using Influenza A Virus As Vector

People infected with rabies virus(RABV)can cause pathogenic encephalitis without intervention,and the fatality rate is close to 100%.This has led to the deaths of tens of thousands of people worldwide in recent years,most of them children,which is a big challenge to public health.The influenza A virus(IAV)genome consists of eight singlestranded negative-sense RNA fragments(v RNA).With the maturity of the influenza A virus reverse genetics operating system,IAV-based recombinant vaccine candidates can be constructed against different pathogens.Here,we rescued two replication-competent recombinant influenza A viruses(r WSN-NA-PHD,r WSN-NS-PHD)whose genomes had embedded fragments of the PHD domain of the rabies virus glycoprotein,one of which was embedded on NA,the other is embedded on NS.The two recombinant influenza A viruses were successfully rescued in 293 T cells and we verified that they were genetically stable,with efficient expression of the PHD protein in infected mammalian cells.When amplified in vitro,the two recombinant viruses had reduced peak titers and attenuated plaque formation compared to wild-type H1N1(A/WSN/33),and were more pronounced on r WSN-NS-PHD.In BALB/c mice,two recombinant influenza A viruses were attenuated in their pathogenicity compared with wild-type H1N1(A/WSN/33)virus and were able to induce mice against rabies virus after two doses of immunization for the production of specific neutralizing antibodies,the induction effect of r WSN-NA-PHD was better than that of r WSN-NS-PHD.Finally,recombinant influenza A virus immunization induces Th1-type cytokines,which may play an important role in immune regulation in the anti-infection immune response.