重组痘苗病毒嵌合巨蛋白的免疫学研究

Study on immunology of recombinant vaccina virus chimeric huge protein

目的 pSFJ1 6与pSFJ38均是以牛痘病毒包涵体 (ATI)启动子和分别串联的 1 6个和 38个p7 5突变型早期启动子为基本构件组成的复合型启动子 (ATI-p7 5)、痘苗病毒复制非必需区血凝素 (HA)基因为侧翼的痘苗病毒表达载体。分别插入编码艾滋病病毒 (HIV - 1 )外膜蛋白env、核心蛋白gag与编码干扰素 (IFNα - 2b)基因 ,观察其表达产物诱导的机体内产生抗体效价的变化规律。方法 利用分子克隆技术构建重组质粒 ,经脂质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vJ1 6env/IFNα - 2b和vJ38gag/IFNα - 2b。免疫小鼠后 ,经间接ELISA分折免疫小鼠血清抗体的OD4 90 值变化。结果 免疫小鼠后体内抗体的OD4 90 值 ,实验组与阴性对照组平均数有显著性差异 (P <0 0 5) ,实验组与阳性对照组之间无显著性差异 (P >0 0 5)。结论 重组痘苗病毒vJ1 6env/IFNα - 2b和vJ38gag/IFNα- 2b免疫小鼠后可诱导机体产生高滴度的抗HIV - 1env、gag与IFNα - 2b巨蛋白抗原的抗体 ,蛋白稀释度与血清抗体OD4 90

Objective pSFJ16 and pSFJ38 were used as expression vector.Both pSFJ16 and pSFJ38 contained ATI promotor and tandem repeat p7.5 mutant stage promotor.HA gene sequence of vaccinia virus was used as reporter gene.Env,gag gene encoding type Ⅰ surface glycoprotein and core protein of AIDS virus(HIV) and encoding interferon α-2b gene of human were inserted into the downstream of the hybid promotor,where HA gene of the vaccinia virus unnecessary areas was the flank of vaccinia virus.This treatise was observed to humoral immunity of organism and effect of antibody induces by vJ16env/IFNα-2b and vJ38gag/IFNα-2b.Homologous recombination was occured in cell.Methods Recombinat plasmids were contructed by molecule clone.The two recombinated expression plasmids and wild-type vaccinia virus were homologously recombinated by liposome transfection in Cos-7 cell with the natural vaccinia virus an selection of the HA plaque as reporter molecule.OD 490 of the serum was observed after mice were immuned.Results There was significantly change between test group and negative control for response of antibody in serum.Positive control was contray( P<0 05,P>0 05 ).Conclusion Recombinant vaccinia virus vJ16env/IFNα-2b and vJ38gag/IFNα-2b could stimulate micebody to produce high level antibody.The more higher effect price of protein is,the lower OD value of antibody is.