Construction And Immunogenicity Of Associated DNA Vaccine Of PRRSV And PCV2Disease And Screening Immunologic Adjuvants

Porcine reproductive and respiratory syndrome (PRRS) and type2of Porcine circovirus (PCV2) were immune suppression diseases. The immune system was against that loss of immunity and even cause other vaccines immune failure when PRRSVor PCV2infection. Thus the diseases of PR, PPV, SF. Paratyphoid. Porcine contagious pleuropneumonia. Edema disease and so on many kinds of viral and bacterial mixed infection and secondary infection had widespread prevalence, lead to the pig group of mortality was higher. It has severely threatened the swine industry, causing tremendous economic losses throughout the pork industry in the world.Therefore developed a safe、efficient and economy applicable of the new vaccine for prevention and treatment the diseases were of great significance.In recent years, with PCV2antigen molecule to PRRSV and the deepening of the research, people gradually found PRRSV virus in the genome ORF5, ORF6and so on some main immunogenicity gene and genome PCV2virus containing the ORF2main structure of protein gene encoding produce antigenic protein can stimulate the body to produce strong antibody response, induction and antibodies produce and the cellular immune mediated the body, prevent virus in hematic disease, eliminate the virus infection plays an important role in the course.In order to further research PRRSV GP5, M and PCV2Cap protein immune function and the experiment in the lab to the nucleic acid vaccine has already set PRRSV on the basis of successful polymerase chain reaction (PCR) pads the Porcine circovirus type2of the main structure ORF2protein gene. On the basis of pIRESneo for eukaryotic expression vector to construct a separate PCV2respectively ORF2gene expression, PRRSV ORF5-ORF6fusion genes and gene and PCV2ORF2、PRRSV ORF5-ORF6fusion gene expression of the restructuring of the nucleic acid vaccine plasmid pIRES-ORF2. pIRES-ORF5-ORF6, pIRES-ORF2-ORF5-ORF6. The RT-PCR、IFA testing results show that the reorganization of the nucleic acid vaccine plasmid purpose gene in in eukaryotic cells get the right to express.In the present study,all animals vaccinated with pIRES-ORF2, pIRES-ORF5-ORF6, pIRES-ORF2-ORF5-ORF6not only developed specific PRRSV or PCV2antibody, as well as T-lymphocyte proliferation response but also induce high level of CTL response. IL-2and IFN-y production against PRRSV antigens has a higher inductive effect on the specific cellular immunity than negative control(P<0.05). In addition, those of adjuvant (QuliA) immunized group were better than those of no adjuvant immunized group on the level of CTL and IFN-y level (P<0.05). These results demonstrated that QuliA has a positive inductive effect on the activation of vaccine-induced virus-specific cellular immune responses in swines, and co-expression of ORF5and ORF6genetic in pIRES-ORF2-ORF5-ORF6has a positive inductive effect on the specific humoral and cellular immunity and could be used as a strategy to develop a new generation of vaccines against PRRSV.