牛病毒性腹泻病毒E2基因优化表达重组卡介苗的免疫试验

Immunogenicity validation of recombinant BCG expressing optimized E2 gene of bovine viral diarrhea disease virus

为评价牛病毒性腹泻病毒(BVDV)E2基因优化表达重组卡介苗(rBCG)对小鼠的免疫效果,分别用已构建的pMV361-E2-1、pMV361-E2-2、pMV361-E2-3、pMV361-E2-4、pMV361-E2-5、pMV361-E2-6重组卡介苗,pMV361空载体,卡介苗,BVDV灭活疫苗和生理盐水对小鼠进行免疫接种,检测各免疫组血清中特异性抗体效价、T淋巴细胞增殖程度和CD4+、CD8+和IL-12的水平,以评价其免疫效果。结果显示,6组重组卡介苗均能够引起小鼠产生BVDV特异性抗体,促进致敏T淋巴细胞增殖,免疫小鼠血清CD4+、CD8+和IL-12的水平均高于对照组,pMV361-E2-1重组卡介苗在诱导小鼠产生BVDV抗体方面要优于其他各组基因重组卡介苗。结果表明,BVDV E2基因不同片段的重组卡介苗均可诱导免疫小鼠产生特异性的体液免疫和细胞免疫,这为BVDV基因工程疫苗的研制奠定了基础。

To evaluate immune efficacy of recombinant BCG（rBCG）, which encodes bovine viral diar- rhea disease virus（BVDV） E2 protein, groups of specific pathogen free（SPF） mice were intraperitoneally injected with the recombinant pMV361-E2-1, pMV361-E2-2, pMV361-E2-3, pMVg61-E2-4, pMV361-E2-5 and pMV361-E2-6, respectively, pMV361, inactivated BVDV vaccine, BCG and sterile normal saline were set as the control groups. The BVDV specific antibody,levels of CD4^＋ ,CD8^＋ ,IL-12 and number of T-lym-phocyte proliferation response were detected and estimated. In result, six groups of rBCG developed not on- ly specific antibodies against BVDV and BCG in mice, but also T lymphocyte proliferation response. Moreo-ver,the levels of CD4^＋, CD8^＋ and IL-12 production against BVDV antigens in the rBCG groups were higher than these in the control groups, and the results showed that the pMV361-E2-1-rBCG was better than the other groups. In conclusion, rBCG with different fragments of E2 gene have a positive inductive effect on the specific humoral and cellular immunity. This study laid the foundation for study on new type gene engineering vaccines against BVDV.