HIV-1gag/IFNα-2b表达产物的生物活性研究  

Study on Immune Response of Mice to Expression Prodution of HIV-1gag/IFNα-2b

作　　者：金宁一[1] 王洪军[2] 王继群[2] 尹旭辉[2] 吴益民[2] 殷震[1] 

机构地区：[1]解放军军需大学研究所，吉林长春130062 [2]沈阳军区军事医学研究所，辽宁沈阳110034

出　　处：《生物技术》2004年第1期14-16,共3页Biotechnology

基　　金：国家自然科学基金项目 (No .39770 66)资助

摘　　要：目的 :将艾滋病病毒核心蛋白 (gag)与干扰素 (IFNα - 2b)融合基因表达的融合蛋白作为免疫原免疫小鼠 ,动态观察小鼠的体液免疫、细胞免疫与CTL应答。方法 :将IFNα - 2b基因片段插入到gag基因的nt5 31位点 ,经脂质体转染与血凝素阴性蚀斑筛选 ,挑出重组痘苗病毒。经SDS -PAGE和Westernblot鉴定表达产物。以小鼠为实验对象 ,用重组痘苗病毒vJ38gag/IFNα - 2b免疫小鼠 ,用ELISA方法检测血清IgG抗体含量。用流式细胞仪测定小鼠外周血CD+ 4 、CD+ 8T淋巴细胞计数。 3H -TdR掺入法检测细胞毒性T淋巴细胞杀伤活性。结果 :血清IgG抗体含量逐渐增高 ,实验组与对照组比较差异有显著性意义 (p <0 .0 5 )。CD+ 4 、CD+ 8T淋巴细胞计数、CTL检测实验组与对照组比较差异均有显著性意义 (p <0 .0 5 )。结论 :重组痘苗病毒vJ38gag/IFNα - 2b能增强小鼠的体液免疫、细胞免疫和CTL应答。IFNα - 2b可以作为免疫佐剂增强机体的免疫状态。Objective:This treatise had observed the Acquired Immunodeficiency Syndrome Virus core protein(gag) and Interferon(IFNα-2b) fusion genes expression product as to immunogen.It consolidates humoral immunity and cellular immunity of organism.Methods:IFNα-2b was inserted to site (nt531) of gag gene,and recombinant vaccinia virus was screened out by liposome transfection and HA-plaque.The expressed product was examined by SDS-PAGE and Western blot.Then the mice were immunized using the purified recombinant virus vJ38gag/IFNα-2b,as control with 0.9% NaCl and vaccine.Dynamics of the serum IgG antibody had examined by ELISA.CD^+_4 and CD^+_8 T- lymphocytes figure of mice peripheral blood lymphocyto was examined by Flow Cytometry (FCM).Cytotoxic T lymphocyte specific killer test was determined by 3H-TdR added.Result:There is significantly change between test group and control for serum IgG antibody(p<0.05).CD^+_4?CD^+_8 T-lymphocytes figure and CTL had significantly change between test group and control(p<0.05).Conclusions:HIV-1gag/IFNα-2b could consolidates humoral immunity and induced mice to producte cellular immunity and could responsed with HIV-1gag positive serum,It possess immungenicity So that it can strengthen immunefunction.

关 键 词：重组痘苗病毒 免疫原 抗体 流式细胞仪 表达产物 生物活性 

分 类 号：R392.2[医药卫生—免疫学]