表达HIV-1 gag-polΔ和gp140TM蛋白复制缺陷型重组腺病毒的构建及免疫效果研究  

Construction of replication-deficient recombinant adenovirus expressing gag-polΔ and gp140TM genes of human immunodeficiency virus in mice

作　　者：刘颖[1] 吴岚[1] 周克明[1] 张旭东[1] 洪坤学[1] 邵一鸣 

机构地区：[1]中国疾病预防控制中心性病与艾滋病预防控制中心病毒与免疫室,北京100050

出　　处：《中华实验和临床病毒学杂志》2004年第2期150-153,共4页Chinese Journal of Experimental and Clinical Virology

基　　金：国家 8 63高技术研究发展计划课题资助(2 0 0 1AA2 15 0 3 1)

摘　　要：目的　构建表达人免疫缺陷病毒Ⅰ型gag polΔ和gp14 0TM基因的非复制型重组腺病毒。方法　首先将HIV 1的gag polΔ和gp14 0TM基因插入穿梭载体 ,再与腺病毒骨架质粒pAdEasy 1共转化E coliBJ5 183,获得重组子。转化 2 93细胞后获得重组病毒。重组腺病毒与DNA疫苗联合免疫小鼠 ,ELISA检测小鼠血清中的抗体。结果　获得两株重组腺病毒vAd gag polΔ和vAd gp14 0TM ,能正确表达Gp14 0TM、Gag蛋白以及经剪切加工的P2 4蛋白 ,与DNA疫苗联合免疫小鼠后能产生高滴度的HIV 1特异性的抗体。结论　成功构建了表达HIV 1结构基因的重组腺病毒 ,能有效诱导HIV 1特异性抗体。Objective Construction of replication-deficient recombinant adenovirus expressing gag-pol and env genes of human immunodeficiency virus (HIV) in mice. Methods gag-polΔ and gp140TM genes were cloned into shuttle vector pAdTrack-CMV respectively,and then the plasmids containing gag-polΔ or gp140TM gene were cotransformed with the backbone of adenovirus into E.coli BJ5183. Transfections of the recombinants were performed to obtain recombinant adenoviruses. Its immunogenicity was evaluated by testing antibody levels of mice primed with DNA vaccines and boosted with recombinant adenoviruses. Results The replication-deficient recombinant adenovirus could express Gp140TM,Gag P55 and P24 proteins correctly. The mice primed with DNA vaccines and boosted with recombinant adenoviruses elicited high titer of HIV-1-specific antibody compared with that inoculated with DNA vaccines only. Conclusion Replication-deficient recombinant adenovirus expressing gag-polΔ and gp140TM can elicit high titer HIV-1-specific antibodies.

关 键 词：基因表达 HIV-1 gag-polΔ gp140TM蛋白 病毒复制缺陷 重组腺病毒 免疫效果 艾滋病 基因组转染 疫苗 

分 类 号：R392[医药卫生—免疫学]