Liu Yang, Guo Yan-Ling, Zhou Shi-Jie, Liu Fei, Du Feng-Jiao, Zheng Xiao-Jing, Jia Hong-Yan, Zhang Zong-De
Laboratory of Molecular Biology for Mycobacteria, Beijing Tuberculosis and Thoracic Tumor Research Institute, Beijing, People's Republic of China.
Clin Vaccine Immunol. 2010 Sep;17(9):1377-80. doi: 10.1128/CVI.00242-10. Epub 2010 Aug 4.
Gamma interferon (IFN-gamma) is a crucial cytokine for protection against Mycobacterium tuberculosis, but the mechanism of IFN-gamma transcription is still unclear. The cyclic AMP (cAMP) responsive element binding (CREB) proteins belong to the bZip (basic leucine zipper) family of transcription factors and are essential for T-cell function and cytokine production. This study focused on the capacity of CREB proteins to regulate IFN-gamma transcription in CD3(+) T cells obtained from tuberculosis (TB) patients and persons with latent tuberculosis infection (LTBI) in China. The electrophoretic mobility shift assay (EMSA), chromatin immunoprecipitation (ChIP), and Western blotting were used to demonstrate the regulatory role of CREB. EMSA (in vitro) and ChIP (in vivo) experiments suggested CREB could bind to the IFN-gamma proximal promoter in persons with LTBI, whereas no binding was detected in TB patients. Western blotting confirmed the expression of CREB proteins, especially serine-133-phosphorylated CREB, was markedly reduced in TB patients compared with persons with LTBI. These results suggested that CREB could promote the transcription and production of IFN-gamma through binding with the IFN-gamma proximal promoter, but the regulatory role of CREB was decreased in tuberculosis patients owing to diminished expression of CREB proteins, which in turn reduced the IFN-gamma production.
γ干扰素(IFN-γ)是抵御结核分枝杆菌的关键细胞因子,但其转录机制仍不清楚。环磷酸腺苷(cAMP)反应元件结合(CREB)蛋白属于转录因子的bZip(碱性亮氨酸拉链)家族,对T细胞功能和细胞因子产生至关重要。本研究聚焦于中国结核病(TB)患者和潜伏结核感染(LTBI)者的CD3(+) T细胞中CREB蛋白调节IFN-γ转录的能力。采用电泳迁移率变动分析(EMSA)、染色质免疫沉淀(ChIP)和蛋白质免疫印迹法来证实CREB的调节作用。EMSA(体外)和ChIP(体内)实验表明,LTBI者中CREB可与IFN-γ近端启动子结合,而TB患者中未检测到结合。蛋白质免疫印迹法证实,与LTBI者相比,TB患者中CREB蛋白,尤其是丝氨酸133磷酸化的CREB的表达明显降低。这些结果表明,CREB可通过与IFN-γ近端启动子结合促进IFN-γ的转录和产生,但在结核病患者中,由于CREB蛋白表达减少,CREB的调节作用降低,进而导致IFN-γ产生减少。