HIV-1C亚型DNA疫苗诱导小鼠免疫反应的研究

 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.

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

The recombinant plasmid was constructed by inserting the HIV-1 clade C gag into a DNA vaccine expression vector-pVAX.Meanwhile the gag gene was inserted into expression vector pGEX-5X-1.The expressed product of gag gene was examined by Western-blot from 293 T cell culture transfected with pVAX-gag and E.coli cell disruption transformed with pGEX-gag.Further,Balb/Cmice were immunized by the recombinant plasmid pVAX-gag,and the cellular immune response of these mice was analyzed by flow cy-tometry and ELISpot assay.The purified recombinant protein Gag was coated in ELISA plate to evaluate hu-moral immune response of these mice.As a result,the HIV-1 clade C DNA vaccine pVAX-gag could effec-tively induce cellular and humoral immune responses in Balb/C mice.These immunological effects showed pos-itive correlation with immunization dose.And the recombinant protein Gag was expressed efficiently inE.col-i.It has antigenicity and can be used for diagnosis of HIV as a helpful antigen.