HIV-1env/IFNα-2b表达产物诱导的淋巴细胞转化与CTL实验  

Lymphocyte transformation and CTLs induced with expression production of HI-1 env/IFNα-2b

作　　者：王洪军[1] 金宁一[1] 年春志[1] 江文正[1] 王继群[1] 魏安明[1] 吴益民[1] 

机构地区：[1]沈阳军区军事医学研究所,沈阳110034

出　　处：《中国人兽共患病杂志》2004年第2期85-87,共3页Chinese Journal of Zoonoses

基　　金：国家自然科学基金项目 (3 9770 66)资助

摘　　要：目的　将艾滋病病毒外膜蛋白与干扰素 (HIV - 1env/IFNα - 2b)融合基因表达的融合蛋白 ,作为免疫原免疫小鼠 ,观察小鼠的免疫功能状态和细胞免疫应答。方法　将IFNα - 2b基因片段插入到env基因的nt8383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vJ16env/IFNα - 2b免疫小鼠 ,以生理盐水和野生型痘苗病毒为对照 ,用MTT法和 3H -TdR掺入法动态检测小鼠脾淋巴细胞对ConA、Lps的反应性与CTL杀伤活性。结果　脾淋巴细胞对ConA、Lps刺激指数与CTL的杀伤活性百分率 ,其实验组与对照组比较差异有显著性意义 (P <0 0 5 )。ConA、Lps刺激指数实验组间比较差异有显著意义 (P <0 0 5 )。结论　重组痘苗病毒vJ16env/IFNα - 2b能增强小鼠脾淋巴细胞的保护性细胞免疫。IFNα - 2b能作为免疫佐剂加强env蛋白的免疫原性 ,提高免疫小鼠的T淋巴细胞免疫能力。Objective To observe the Acquired Immunodeficiency Syndrome virus exmembrane protein and interferon(HIV-1 env/IFNα-2b)fusion genes expression product as to gene vaccine,it consolidates immune function and cellular immunity of organism.Methods IFNα-2b gene was inserted to site(nt8383)of env 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 vJ16env/IFNα-2b,as control with 0.9% NaCl and vaccinia virus.ConA and Lps's response of T-lymphocytes in spleen had been examined by MTT 570nm.Cytotoxic T lymphocyte specific killer test was determined by 3H-TdR added.Results There is significantly chang between test group and control for ConA and Lps's response of T-lymphocytes and CTL in spleen(P<0.05).Conclusion HIV-1 env/IFNα-2b can consolidates immune function and induced mice to produce cellular immunity,IFNα-2b can strengthen immunogenicity of env protein and increase capability of immune cell of immunized mice.

关 键 词：重组痘苗病毒 免疫原性 CTL 淋巴细胞转化 免疫功能状态 细胞免疫应答 

分 类 号：R373.9[医药卫生—病原生物学]