HIV-1C亚型DNA疫苗诱导小鼠免疫反应的研究

Cellular and humoral immune responses induced by HIV-1 clade C DNA vaccine

构建含中国流行株HIV-1 C亚型核心蛋白gag基因的重组质粒pVAX-gag,并在体外进行了表达与鉴定.同时构建了含此gag基因的原核表达质粒pGEX-gag,表达纯化并鉴定重组蛋白Gag.以质粒pVAX-gag免疫Balb/C小鼠后,用ELISpot和流式细胞仪检测其细胞免疫反应.再以纯化后的重组蛋白Gag作为包被抗原,用ELISA检测其体液免疫反应.结果显示重组质粒pVAX-gag免疫小鼠后可有效地诱导机体产生细胞免疫和体液免疫反应,且免疫剂量和免疫效果存在一定的正相关性.重组原核表达质粒pGEX-gag的表达产物能与抗p24单克隆抗体发生特异性反应,可用于抗HIV抗体检测.

The recombinant plasmid was constructed by inserting the HIV-1 elade C gag into a DNA vaccine ex- pression veetorypVAX. Meanwhile the gag gene was inserted into expression vector pGEX-5X-1. The expressed prod- uct of gag gene was examined by Western-blot from 293 T cell culture transfeeted with pVAX-gag and E. eoli cell disruption transformed with pGEX-gag. Further, Balb/C mice were immunized by the recombinant plasmid pVAX-gag, and the cellular immune response of these mice was analyzed by flow eytometry and ELISpot assay. The purified recombinant protein Gag was coated in ELISA plate to evaluate humoral immune response of these mice. As a result, the HIV-1 elade C DNA vaccine pVAX-gag could effectively induce cellular and humoral immune responses in Balb/C mice. These immunological effects showed positive correlation with immunization dose. And the recombinant protein Gag was expressed effieiently in E. coll. It has antigenieity and can be used for diagnosis of HIV as a helpful antigen.