HIV-1env、gag与IFNα-2b基因融合蛋白的免疫学研究

Study on Immunity of Expression Protein of HIV-1env ,gag and IFNα-2b Fusion Genes in Eukaryotic Cell

目的将编码艾滋病病毒(HI V-1)外膜蛋白的env基因、核蛋白的gag基因与编码干扰素(IFNα-2b)基因分别插入pSFJ16与pSFJ38真核表达载体,观察表达产物诱导机体产生细胞免疫的变化规律。方法利用分子生物学技术构建重组质粒,经质脂体转染与野生型痘苗病毒在Cos-7细胞内同源重组,筛选、纯化获得重组痘苗病毒vJ38gag/IFNα-2b和vJ16env/IFNα-2b。免疫小鼠后,检测小鼠外周血与脾淋巴细胞对ConA及Lps的反应性,用流式细胞仪测定小鼠脾细胞CD4+、CD8+,细胞计数。结果外周血与脾淋巴细胞对ConA及Lps的反应性,实验组与对照组比较有显著差异(P<0.05)。CD4+T淋巴细胞与对照组比较有显著差异(P<0.05)。CD8+T淋巴细胞与对照组比较无显著差异(P>0.05),但呈增高趋势。结论重组痘苗病毒能诱导小鼠产生较强的细胞免疫。重组痘苗病毒能在体外与HI V-1env和HI V-1gag阳性血清发生特异性反应,具有免疫原性和免疫反应性。

Objective To observe the immune reaction that induced by the Expression of HIV-1 env, gag and IFNα-2b Fusion Genes in Eukaryotic Cell in mice body cell. Methods By liposome transfection, four recombinant virus strain, vJ16env/IFNα-2b,vJ38gag/IFNα-2b,vJ16env and vJ38gag were obtained by HA- plaque screening. The mices were immunized with the four strain recombinant vaccinia virus, 1 × 10^7/mice. OD value of HIV-1 env and HIV-1 gag antibody was determined. CD4^＋ . CD8^＋ T cell figure of mice spleen was examined by Flow Cytometry （FCM）. Results There were signifcant differences （P 〈 0.05）, btween OD value. CD4 ^＋ T cell counts and CD4^＋/CD8^＋ ratio of the group and the contrive group. Study demostrated： The its immungenicity and immunereposibility was proved and may be significant in further re- search in this area. Conclusion Recombinant vaccinia virus can induce powerly cellular immune reaction by vaccinating into mice body and have reaction to serum with HIV-1 env,gag in test,Its immungenicity and immunereposibility was proved.