腺病毒介导的白细胞介素-12表达对丙型肝炎病毒包膜2基因免疫的调节  

Adenovirus mediated expression of interleukin 12 regulating hepatitis C virus E2 gene immunization induced immune response

作　　者：吴朝栋[1] 李红桂[1] 陶其敏[1] 魏来[1] 

机构地区：[1]北京大学人民医院,北京大学肝病研究所,北京100044

出　　处：《中华肝脏病杂志》2004年第10期620-622,共3页Chinese Journal of Hepatology

基　　金：国家十五科技攻关计划(2001BA705B06);国家自然科学基金(30170844);中华医学基金专项人才基金

摘　　要：目的 观察腺病毒介导表达白细胞介素-12(IL-12)对调节丙型肝炎病毒(HCV)包膜基因2(E2)诱导免疫应答的影响。 方法 以NIH 3T3细胞表达HCV E2糖蛋白,纯化后用于酶免疫分析法检测抗E2抗体;以表达HCV E2糖蛋白的SP2/0细胞51Cr释放法检测细胞毒性T淋巴细胞(CTL)应答;以293细胞繁殖表达重组IL-12亚单位p35和p40的腺病毒ADIL12。6-8周龄BALB/C鼠右后肢股四头肌注射表达HCV E2的质粒,同时经腹腔注射ADIL12。分别于2、3、4周尾静脉采血检测抗E2抗体,4周处死动物分离脾细胞检测CTL应答。 结果 表达HCV E2的基因免疫可有效诱导特异性抗E2体液免疫应答。腹腔注射后,外源性IL-12微量表达。微量表达的外源性白细胞介素-12不影响特异性抗E2体液免疫应答。同时,显著增强特异性CTL应答的作用。 结论 腺病毒微量表达的IL-12对HCV E2基因免疫诱导的特异性CTL应答具有调节作用。Objective To observe the regulating effect of hepatitis C virus (HCV) envelop (E) 2 gene immunization-induced immune responses by adenovirus mediated interleukin 12 (IL-12). Methods HCV E2 protein was expressed and purified from NIH 3T3 and then used as an antigen to detect antibodies against HCV E2. With 51Cr release, SP2/0 expressing HCV E2 was used as target cell to detect specific cytotoxic T lymphocytes (CTL) response; adenovirus recombined IL-12 was propagated by 293 cell. HCV E2 recombinant and adenovirus recombined IL-12 were injected into the quadriceps femoris muscles and abdominal cavities of 6-8 weeks old BALB/C mice. Sera were collected at 2, 3, and 4 weeks and detected for antibodies for E2. Spleen cells isolated at 4 weeks were analyzed for specific CTL response. Results It was found that expression of IL-12 at an undetectable level did enhance HCV E2 gene immunization-induced CTL activity and there was no effect on its hormonal immune response. Conclusions Using adenovirus to express interleukin 12 was helpful for regulation of HCV E2 gene immunization-induced immune response. Combined HCV E2 and IL-12 can render a strong anti-HCV CTL activity and may be of use in the development of HCV gene vaccine in the future.

关 键 词：表达 抗E 白细胞介素-12 腺病毒介导 IL-12 丙型肝炎病毒 量表 糖蛋白 3T3细胞 SP2/0细胞 

分 类 号：R392[医药卫生—免疫学] R512[医药卫生—基础医学]