HBsAg与GM-CSF基因双表达载体的构建及其免疫效果  

Construction and Immune Effect of Recombinant Plasmid for Co-expression of HBsAg and GM-CSF Genes

作　　者：冷梅[1] 闫刚[2] 迟春萍[1] 韩顺子[1] 吴晓娟[1] 赵大鹏[1] 汪春义[1] 郑全莉[1] 李鑫[2] 李雨桐[1] 张秀霞[1] 姜崴[1] 

机构地区：[1]长春生物制品研究所,长春130062 [2]长春市卫生局卫生监督所,长春130033

出　　处：《中国生物制品学杂志》2008年第5期401-404,共4页Chinese Journal of Biologicals

摘　　要：目的构建乙型肝炎表面抗原基因(HBsAg)与GM-CSF基因的双表达载体,提高乙肝病毒DNA疫苗的免疫效果。方法将微小病毒内部核糖体进入位点(IRES)基因克隆到质粒pVAXⅠ多克隆位点,再将HBsAg基因和GM-CSF基因依次克隆到IRES的上、下游多克隆位点处,构建双表达载体pHIG。将pHIG转染到COS-7细胞中,检测瞬时表达情况,并用双表达质粒pHIG免疫BALB/c小鼠,检测免疫后小鼠血清中抗-HBs及其脾脏T细胞表面CD4+、CD8+分子的数量。结果在转染pHIG质粒的COS-7细胞上清液中有HBsAg的表达,pHIG双表达质粒组比pVAX/HBs组抗体产生时间提前2周,抗体阳转率提高2倍,小鼠脾脏T细胞表面CD4+分子数量及CD4+/CD8+值均高于pVAX/HBs组。结论HBsAg与GM-CSF基因双表达质粒能引起小鼠特异性免疫应答,并可提高免疫效果。Objective To construct a recombinant plasmid for co-expression of HBsAg and GM-CSF genes and enhance the immune effect of hepatitis B virus DNA vaccine. Methods Clone parvovims internal ribosomal entry site （IRES） gene into plasmid pVAX I , then clone HBsAg and GM-CSF genes upstream and downstream to IRES. Transfect the constructed co-expression vector pHIG to COS-7 cells and determine the transient expression of HBsAg. Immunize BALB/c mice with pHIG and determine the anti-HBs in sere as well as CD4^＋ and CD8^＋ molecules on the surface of T cells in spleens of mice. Results HBsAg was expressed in the culture supernatant of COS-7 cells transfected with pHIG.The anti-HBs in sera of mice immunized with pHIG was produced 2 weeks earlier than that with pVAX/HBs,and the antiHBs positive conversion rate of the former was 2 times higher than that of the latter. Both CD4 ^＋ molecule count and the ratio of CD4 ^＋ to CD8 ^＋ on surface of T cells in spleens of mice immunized with pHIG were significantly higher than those with pVAX/HBs. Conduslon The co-expression vector for HBsAg and GM-CSF induced soecific immune resoonse in mice and enhanced immune effect.

关 键 词：乙型肝炎表面抗原 DNA疫苗 粒细胞-巨噬细胞集落刺激因子 内部核糖体进入位点 

分 类 号：Q78[生物学—分子生物学] R373.21[医药卫生—病原生物学]