结核分枝杆菌抗原Ag85B优势诱导活动性结核患者Th1型细胞免疫反应研究  

Enhancement of Th1 Type Cellular Immune Response in Active TB Patients Induced by Ag85B Mycobacterium Tuberculosis Antigens

作　　者：王洁玲[1] 唐余燕[1] 张毅[1] 汤正好[1] 臧国庆[1] 余永胜[1] 

机构地区：[1]上海交通大学附属第六人民医院感染科,200233

出　　处：《医学研究杂志》2016年第3期77-82,共6页Journal of Medical Research

基　　金：上海市教育委员会科研创新项目(1522013)

摘　　要：目的观察Ag85B抗原在诱导活动性结核患者细胞免疫应答中的作用。方法选取60例活动性结核患者,分离其外周血单个核淋巴细胞(PBMC),加Ag85B抗原刺激培养72h,ELISA法检测上清液中的IFN-γ、IL-2、IL-4及IL-10细胞因子浓度,流式细胞术(FCM)检测CD4^+T淋巴细胞内的细胞因子IFN-γ水平,定量PCR及Western blot法检测PBMC细胞转录因子T-bet、GATA-3的表达。结果检测活动性结核患者外周血单个核淋巴细胞(PBMC)培养上清液,Ag85B蛋白实验组Th1型细胞因子IFN-γ、IL-2浓度明显高于结核菌素(PPD)组(P<0.05),而Th2型细胞因子IL-4、IL-10水平与PPD组相比,差异无统计学意义(P>0.05),以上两组细胞因子水平均明显高于空白对照组(P<0.05)。流式细胞法检测CD4+T胞内细胞因子IFN-γ水平,Ag85B蛋白实验组高于PPD组及空白对照组(P<0.05)。定量PCR及Western blot法检测,T-bet水平Ag85B组高于PPD组(P<0.05),而GATA-3水平差异无统计学意义(P>0.05)。结论结核杆菌抗原Ag85B可以通过上调Th1型转录因子T-bet诱导优势Th1型免疫反应,为结核病的预防及治疗提供理论基础。Objective To observe whether the Ag85B antigen could enhance Th1-type cellular immune response in active TB patients and study the mechanism. Methods In 60 patients with active TB, peripheral blood PBMC cell were Separated,and they were stimulated with the antigen Ag85B, purified protein derivative（PPD） and Phosphate Buffered Saline（PBS）, respectively. The levels of Th1/Th2 type cytokines（IFN-γ, IL-2 and IL-4, IL-10） secreted by PBMC cells were analyzed by ELISA. Flow cytometry （FCM） was used to detecte the CD4＋T cell cytokines IFN-γ. The expression of PBMC cells specific T-bet and GATA-3 were detected by realtime quantitative polymerase chain reaction and Western blot analysis. Results Antigen Ag85B significantly increased the Th1-type cytokine（IFN-γ, IL-2） in active TB patients compared with PPD （P〈0.05）, while Th2 cytokines IL-4, IL-10 level compared with PPD group,had no statistically significant difference （P〉0.05）. By applying the method of flow cytometry,We detected intracellular cytokines IFN-γ levels of CD4＋ T lymphocyte number which was higher than PPD group and normal control group （P〈0.05）. The expression of T-bet was significantly up-regulated in the Ag85B group compared with the control groups （P〈0.05）. Moreover, the expression of GATA-3 had no significant difference between Ag85B group and PPD group （P〉0.05）. Conclusion Ag85B mycobacterium tuberculosis antigens by raising Th1 type transcription factor induced advantage T-bet Th1 immune response.

关 键 词：活动性结核 Ag85B抗原 Th1型细胞免疫 

分 类 号：R5[医药卫生—内科学]