重组痘苗病毒表达人免疫缺陷病毒中国株B’亚型Gag蛋白及免疫效果观察  

The Expression of gag Gene of Chinese HIV-1 Strain(RL_(42),clade B′)in Vaccinia Virus and Its Immune Potency in Mice

作　　者：刘锦彦[1] 陶欣 洪坤学 吴岚 刘颖 朱家鸿[1] 邵一鸣 

机构地区：[1]武汉生物制品研究所,湖北430060 [2]国家艾滋病参比实验室,北京100050

出　　处：《病毒学报》2002年第1期9-14,共6页Chinese Journal of Virology

基　　金：国家 8 6 3高技术研究发展计划 ( 86 -10 2 -0 7-0 2 -0 5 )

摘　　要：为筛选用于我国HIV疫苗的候选基因 ,利用痘苗病毒载体构建表达HIV - 1B′亚型中国流行株RL4 2 的Gag蛋白的重组病毒rVV -RL4 2 gag ,并免疫BALB/c小鼠 ,检测其诱导的特异性抗体及中和抗体 ,同时检测其诱导的特异性CTL及与中国流行株C(B′/C重组 )亚型和国际流行株A亚型之间的CTL交叉反应。结果显示 :重组病毒能很好地表达Gag蛋白 ,它具有与 7株单克隆抗体结合的抗体表位 ;电镜下可观察到Gag蛋白形成的颗粒。rVV -RL4 2 gag免疫小鼠后能诱导产生高滴度的特异性抗体和CTL ,抗体具一定的中和活性 ,且检测到与C(B′/C重组 )亚型和国际流行株A亚型之间的CTL交叉反应。这些结果表明 ,rVV -RL4 2 gag具有良好的免疫原性 ,可进一步构建表达HIVB′亚型中国流行株RL4 2 的Gag蛋白的重组痘苗病毒作为候选疫苗。A recombinant vaccinia virus containing the gag gene of Chinese epidemic HIV-1 strain(RL 42 ,clade B′), under the control of vaccinia virus promoter p7.5,was constructed by inserting the gag gene into the TK region of vaccinia virus. Through ELISA and Western blot assays,it was shown that the recombinant rVV-RL 42 gag could successfully express the HIV p55.Gag protein had the epitopes capable of binding seven monoclonal antibodies,and could assemble into particles that could be detected by electron microscope.The recombinant vaccinia virus rVV-RL 42 gag could induce humoral and CTL immune responses after inoculating BALB/c mice, the specific and neutralization antibodies could be detected.The CTL induced by recombinant virus could cross-react with clade A,B′and C(B′/C)peptide epitopes.The results suggested that recombinant vaccinia virus containing the gag gene of Chinese HIV-1 strain(RL 42 ,clade B)can be constructed as a candidate HIV-1 vaccine of China.

关 键 词：人免疫缺陷病毒I型 HIV-1 重组痘苗病毒 GAG蛋白 细胞毒性T淋巴细胞 抗性 中和抗性 免疫效果 

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