融合表达牛疱疹病毒Ⅰ型VP22基因和O型口蹄疫病毒P12A3C基因的DNA疫苗的构建及其免疫应答

Construction and immune response of DNA vaccine recombinant PRV co-expressing VP22 gene of BHV-Ⅰ and P12A3C gene of O type FMDV

用小鼠模型评价融合表达牛疱疹病毒Ⅰ型VP22基因和O型口蹄疫病毒P12A3C基因的DNA疫苗和不同免疫策略的免疫应答。用PCR方法扩增牛疱疹病毒Ⅰ型VP22基因和O型口蹄疫病毒P12A3C基因,分别克隆到pMD18-T载体并测序验证正确后将其克隆到质粒pcDNA的相应位点获得质粒pcDNA-VP22-P12A3C。然后将BALB/c小鼠分成7组进行免疫。结果表明,DNA疫苗pcDNA-VP22-P12A3C诱导的细胞免疫水平超过了灭活疫苗,DNA疫苗与灭活疫苗联合免疫组体液免疫水平接近灭活疫苗组而细胞免疫水平远高于灭活疫苗组,为进一步研究VP22和P12A3C融合表达的基因工程疫苗奠定了基础。

To evaluate the immune effect of the DNA vaccine recombinant PRV co expressing P12A3C protein of foot and mouth disease virus with VP22 of bovine herpes virus I in different immune strategy in mice. The complete coding regions of FMDV P12A3C gene and BHV-1 VP22 gene having no stop code were amplified by PCR from the plasmids containing P12A3C gene or VP22 gene, respectively. And P12A3C gene and VP22 gene were cloned into pMD18-T vector and sequenced correctly,then the fragments were inserted into plasmid pcDNA to got the plasmid pcDNA-VP22-P12A3C. Fifty-six BALB/c mouse were separated into seven groups and immuned according to the strategy. Humoral-mediated immune responses induced by pcDNA-VP22-P12A3C DNA vaccine in combination with inactivated vaccine were no different with the inactivated vaccine alone. But cell-mediated immune responses showed that pcDNA-VP22-P12A3 induced stronger lymphocyte proliferative responses than inactivated vaccine, which established the foundation of the research in DNA vaccine.