中国流行株HIV-1gag和hIL-2/hIL-6核酸疫苗构建与实验免疫研究  

Construction of DNA vaccine with HIV-1 gag and hIL-2/hIL-6 and its immunological study

作　　者：秦云龙[1] 金宁一[2] 罗坤[1] 李凡[1] 王宏伟[2] 王莉馨[2] 张永生[1] 殷震[2] 

机构地区：[1]白求恩医科大学病原生物学教研室,吉林长春130021 [2]解放军军需大学研究所病毒室,吉林长春130062

出　　处：《免疫学杂志》2001年第1期5-8,共4页Immunological Journal

基　　金：国家自然科学基金(3977066);国家杰出青年基金(39770661)资助项目

摘　　要：目的探讨中国流行株 HIV- 1gag与 h IL - 2 /h IL - 6共表达重组核酸疫苗质粒的免疫效果。方法以核酸疫苗质粒 p IRES1neo为表达载体 ,构建重组核酸疫苗质粒 p IRES1- gag、p IRES1- gag- h IL- 2、p IRES1- gag- h IL- 6 ,通过间接免疫荧光试验、Dot- EL ISA检测 gag/h IL - 2 /h IL - 6基因的表达产物。另将此重组核酸疫苗质粒免疫 Balb/c小鼠 ,进行淋巴细胞转化试验、CD4+ 、CD8+ T淋巴细胞数量测定、细胞毒性 T淋巴细胞 (CTL )特异性杀伤作用检测及血清抗体检测 ,结果构建的重组质粒转染 BHK细胞后可表达目的基因 ,免疫小鼠后可有效地刺激淋巴细胞增殖、诱导特异性 CTL 反应 ,当和 h IL- 2 /h IL- 6共表达时免疫效果更加显著。讨论与 Gag蛋白共表达的 h IL- 2 /h IL- 6能够进一步增强免疫鼠的细胞免疫与体液免疫水平 ,构建的重组质粒为 HIV-ObjectiveTo study the immunological effects of recombinant HIV 1 DNA vaccine plasmids in mice MethodsTwo DNA vaccine recombinant plasmids pIRES1 gag hIL 2,pIRES1 gag hIL 6 were constructed by inserting the HIV 1 gag and hIL 2/hIL 6 gene into a DNA vaccine expression vector(pIRES1neo). The expressed product of gag/hIL 2/hIL 6 gene was examined by Dot ELISA and indirect immunofluorescent assay Immune response of Balb/c mice immunized by the two DNA vaccine recombinant plasmids was examined by lymphocyte transformation test(LTT),cytotoxic T lymphocyte specific killer test,quantity change of T lymphocyte (CD4 +, CD8 +) and dynamics of specific serum IgG antibody ResultsHigh effective expression was found in BHK cells transfected with pIRES1 gag hIL 2,pIRES1 gag hIL 6 Recombinant plasmids could effectively induce the proliferation of effector lymphocytes and the generation of specific lytic activity in vitro The effects were more significant when co expressed with human interleukins gene ConclusionCytokine hIL 2/hIL 6 can enhance both humoral and cellular immunity in immunized mice Constructed recombinant plasmids might provide a basis for the development of HIV 1 DNA vaccine

关 键 词：HIV-1 GAG蛋白 白细胞介素-2 白细胞介素-6 

分 类 号：R392-33[医药卫生—免疫学]