结核分枝杆菌早期分泌抗原靶-6（ESAT-6）对人THP-1单核细胞IL-12产生的影响

摘要/Abstract

摘要： 目的研究结核分枝杆菌早期分泌抗原靶-6（ESAT-6）对人THP-1单核细胞IL-12产生的影响及其作用的细胞信号传导机制。方法应用亲和层析方法纯化大肠埃希菌表达重组的结核分枝杆菌ESAT-6抗原，采用ELISA法检测不同浓度ESAT-6刺激人THP-1单核细胞IL-12产生的影响，以及对LPS刺激的反应，应用各种细胞信号传导通路抑制剂来探讨ESAT-6诱导单核细胞IL-12产生相关可能的信号通路。结果结核分枝杆菌ESAT-6分泌抗原在2.5～10g/ml浓度范围能依赖性地刺激人THP-1单核细胞产生IL-12（p70 及其亚单位p40）。细胞信号传导通路JNKMAPK的选择性抑制剂SP600125能促进ESAT-6刺激单核细胞IL-12p40产生，而信号通路PKR抑制剂2-AP有显著性抑制其作用。结论ESAT-6抗原诱导人THP-1单核细胞IL-12产生，JNK MAPK及PKR信号通路可能参与了此过程的调控。

关键词: 分枝杆菌, 结核, 抗原, 细菌, 细菌蛋白质类, 单核细胞, 白细胞介素12

Abstract: ObjectiveTo study the effect of Mycobacterium tuberculosis 6-kilodalton early secreted antigenic target (ESAT-6) on interleukin 12 (IL-12) production from human THP-1 monocytes. MethodsExpression and purification of ESAT-6 in E.Coli. Recombinant ESAT-6 was purified by nickel-nitrilotriacetic acid metal affinity chromatography under native conditions. Human monocytes were treated with ESAT-6, in present of LPS or not at different culture periods or dose. Supernatants were collected for IL-12 detection by ELISA. Signaling pathway inhibitors was used to find the signaling pathway of ESAT-6. ResultsMycobacterium tuberculosis ESAT-6 antigen dose-dependently induced IL-12 release from THP-1 monocytes; Lipopolysaccharide (LPS) had synergic effect on ESAT-6 induced IL-12 production from THP-1 monocytes.1； The enhancement effect of ESAT-6 on IL-12 production was promoted by selective inhibitor of JNK MAPK, SP600125, while it was inhibited by PKR selective inhibitor, 2-AP. ConclusionESAT-6 may induce monocytes activation and enhance IL-12 release. This may have important significance of protection in the early period of infection.

Key words: Mycobacterium tuberculosis, Antigens,bacterial, Bacterial proteins, Monocytes, Interleukin-12

刘耀婷,胡忠义,冯永红,刘忠华,拾莉. 结核分枝杆菌早期分泌抗原靶-6（ESAT-6）对人THP-1单核细胞IL-12产生的影响[J]. 中国防痨杂志, 2009, 31(7): 384-388.

Liu Yaoting¹，Hu Zhongyi²，Feng Yonghong²，Liu Zhonghua, Shi Li. Effect of Mycobacterium tuberculosis 6-kilodalton early secreted antigenic target (ESAT-6) on IL-12 production from THP-1 monocytes[J]. Chinese Journal of Antituberculosis, 2009, 31(7): 384-388.

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