Construction and immunogenicity evaluation in mice of recombinant pseudorabies viruses expressing porcine reproductive and respiratory syndrome virus GP5(GP5m),M protein and porcine IL-18
To obtain an effective recombinant live vector vaccine for the prevention of porcine reproductive and respiratory syndrome(PRRS)and porcine pseudorabies(PR),GP5,modified GP5(GP5m),and M gene of the PRRS virus(PRRSV)infetious clone ATCC-VR2332 and porcine IL-18 gene was inserted into the TK gene site of rPRV-gE-/TK-/GFP+deletion strain by homologous recombination,five recombinant pseudorabies viruses(rPRVs)were constructed,respectively,namely rPRV-gE-/TK-/GP5m+,rPRV-gE-/TK-/GP5m+/IL-18+,rPRV-gE-/TK-/GP5+/M+,rPRV-gE-/TK-/GP5+/M+/IL-18+,and rPRV-gE-/TK-/IL18+.Immunoblotting results showed that the five rPRVs could express PRRSV GP5,GP5m,M protein or porcine IL-18.Subsequently mice were immunized with these five rPRVs,respectively,rPRV-gE-/TK-/GP5+/M+,rPRV-gE-/TK-/GP5+/M+/IL-18+and rPRV-gE-/TK-/GP5m+/IL-18+could produce anti-PRRSV and PRV specific neutralization antibodies,which could stimulate the proliferation of CD3+,CD4+,and CD8+T lymphocytes and the expresstion of IFN-y,IL-4,and IL-10 cytokine.In summary,rPRV-gE-/TK-/GP5+/M+,rPRV-gE-/TK-/GP5+/M+/IL-18+,and rPRV-gE-/TK-/GP5m+/1L-18+could effectively induce specific systemic immune response against PRRSV and PRV in mice,which might lay the foundation for the development of the recombinant live vector vaccine for effectively prevention of PRRS and PR.
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