中国流行株HIV-1B亚型Gag重组鸡痘病毒构建及其免疫原性

Construction of Recombinant Fowlpox Virus of HIV-1 Capsid Protein Gag and Its Immunogenity

在以鸡痘病毒 ( FPV) 2 82 E4 株为基础构建的重组表达质粒 p UTAL的 ATI-P7.5复合启动子下游 ,插入编码中国流行株 HIV-1 B亚型核心蛋白 gag基因 ,构建了重组表达质粒 p UTALG。重组质粒与 FPV共转染 CEF细胞 ,进行了同源重组。通过 BUd R加压筛选 ,X-gal染色 ,获得重组鸡痘病毒 v UTALG。 Westernblot检测结果表明 ,重组病毒表达了 Gag蛋白 ,裂解后其相对分子质量分别为 5 5× 1 0 3 、4 8× 1 0 3 、2 4× 1 0 3 和1 7× 1 0 3 。以重组病毒 v UTALG免疫小鼠 ,与 FPV对照比较 ,小鼠脾 T淋巴细胞 CD4 + T细胞数和 CD4 + /CD8+值均有上升趋势 ,Con A、LPS诱导的脾细胞增殖能力增强 ,并能诱导 CTL和 HIV-1特异的血清抗体反应 。

The gag gene of HIV-1 was inserted at downstream of the combined promoter in p UTAL,a expressing plasmid was constructed.By transfecting the plasmid into CEF cells pre-infected with FPV 2 82 E4 strain,a recombinantvirus v UTALG was obtained by selecting in the presence of BU- d R and subsequentblue plaque screening.Western blot showed that the recombinant virus expressed the Gag protein in the infected CEF cells.Mice were immunized with the recombinantvirus.The num- ber of CD4 + cells from splenic lymphocytes increased and the proliferation response of splenocytes to Con A,LPSand CTL cytotoxicactivity elevated markedly with the HIV-1 -specific antibody responseto be induced.The results showed the recombinant virus could increase cellular and humoral immunity response in mice.