Study On Preparation And Immune Efficacy Of Inactivated Vaccines Of Chinese Giant Salamander Ranavirus (CGSRV)

This study firstly carried out with preparation of inactivated vaccine, screening adjuvant, secondly determination of serum lysozyme activity, ACH50 activity, antibody titers and expression of major histocompatibility complex class (MHC) I gene, MHCⅡene and Interferon a-inducible protein 6 (IFI 6) gene after immuning the prepared vaccine, and finally determination of the relative percent survival rate (RPS). By studying the immune related indicators and the protective effect of Chinese giant salamander after immuning the prepared vaccine, designed to provide a theoretical basis for further study to control the disease of Chinese giant salamander ravavirus.CGSRV was inactivated by β-propiolactone (BPL), and used to prepare the inactivated vaccine, inactivated vaccine with astragalus polysaccharide (APS) adjuvant and inactivated vaccine with ISA763A adjuvant, these three vaccines are studied in this paper.Vaccines were inoculated intraperitoneal to Chinese giant salamander with a dose of 0.3mL after sterility test and safety inspection. Blood was collected after immunization in 10d,20d,30d,40d, and detected the changes of immune indicators, including lysozyme activity, ACH50 activity, antibody titer in serum, and relative expression of MHC I gene, MHC ⅡA gene and IFI 6 gene in blood.40 days after immunization,15 Chinese giant salamanders challenge test was carried out and the relative percent survival rates was measured, the other 5 Chinese giant salamanders continued to test antibody titer. The results showed that inactivated vaccine and inactivated vaccine with adjuvant can be varying degrees to improve the body’s immune response, and antibody titer, ACH50 activity, relative expression of MHC I gene, MHC ⅡA gene and IFI 6 gene increased in Chinese giant salamanders, three vaccine had different effect on lysozyme activity except inactivated vaccine with APS adjuvant. The data of immune indicators showed that the inactivated vaccine could enhance the immunity of Chinese giant salamader, the immune enhancement was improved at both ISA763A adjuvant and APS adjuvant. The RPS of inactivated vaccine with ISA763A group was 85.71%, inactivated vaccine with APS group was 71.43%, inactivated vaccine group was 64.29%. ISA763A adjuvant and APS adjuvant separately improved RPS at 21.41% and 7.14%, and due to ISA763A adjuvant’s immune better, it is recommended ISA763A as adjuvant of CGSRV inactivated vaccine.The results mentioned above reveal that CGSRV inactivated vaccine not only improved the specific immune involving to produce antibody and to induce MHC I gene, MHC ⅡA gene express, but also improved the non-specific immune involving to improve the ACH50 activity and to induce IFI 6 gene express. Both ISA763A adjuvant and APS adjuvant improved immune efficacy of CGSRV inactivated vaccine, and the immune enhancement was better of ISA763A adjuvant.