含HIV-1 gag、gag V3基因的重组腺病毒伴随病毒的构建及其免疫原性的研究  

Construction of Recombinant Adeno-Associated Virus Which Expressed HIV-1 gag,gagV3 and the Preliminary Study of Their Antigenicity

作　　者：刘雁征[1] 吴小兵[1] 周玲[1] 伍志坚[1] 侯云德[1] 曾毅[1] 

机构地区：[1]中国预防医学科学院病毒学研究所,北京100052

出　　处：《病毒学报》2001年第4期328-331,共4页Chinese Journal of Virology

摘　　要：将HIV - 1中国株 42 (B亚型 )的 gag 基因及 gag与 gp12 0V3区的嵌合基因 gagV3插入腺病毒伴随病毒(AAV)表达载体pSNAV质粒中 ,构建重组质粒 pSNAV gag及pSNAV gagV3;采用脂质体转染的方法分别将重组质粒转入BHK细胞 ,G418筛选得到转入重组质粒并能表达外源基因的细胞系 ,命名为BHK gag及BHK gagV3。用具有重组腺病毒伴随病毒 (rAAV)包装功能的一种重组单纯疱疹病毒 (rHSV)分别感染这两株细胞系 ,纯化后得到rAAV ,电镜观察可见到大量实心病毒颗粒 ,核酸杂交检测重组病毒滴度达到 10 12 病毒颗粒 /ml,重组病毒感染2 93细胞 ,ELISA检测有 gag 及 gagV3基因的表达。用重组病毒免疫Balb/C小鼠 ,检测抗体及细胞免疫水平 ,证明重组病毒可以在小鼠体内诱导产生细胞及体液免疫。The gag and gagV3 gene of the Chinese HIV-1 strain 42(subtype B) were cloned into the adeno-associated virus(AAV) vector pSNAV,the recombinant pSNAV-gag?pSNAV-gagV3 were transfected into BHK cells by using Lipofectamine.The G418-resistant cells,BHK-gag and BHK-gagV3,were obtained.These cells were infected with recombinant HSV which has function of packaging the recombinant AAV (rAAV).After purification,we obtained high-titer rAAV(10 12 particles/ml) which expressed HIV-1 gag and gagV3.Balb/C mice were immunized with rAAV,the rAAV could induce HIV-1 specific cell-mediated and humoral immunity.These results showed a hope of developing rAAV as HIV vaccine.

关 键 词：人免疫缺陷病毒1型 HIV-1 重组腺病毒伴随病毒 细胞毒性T淋巴细胞 CTL活性 

分 类 号：R373.9[医药卫生—病原生物学] R512.91[医药卫生—基础医学]