Effect of Mycobacterium tuberculosis 6-kilodalton early secreted antigenic target (ESAT-6) on IL-12 production from THP-1 monocytes

Abstract

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

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.

References

[1] Harboe M，Oettinger T，Wiker HG，Rosenkrands I，Andersen P.Evidence for occurrence of the ESAT-6 protein in Mycobacteria tuberculosis and virulent Mycobacteria bovis and for its absence in Mycobacteria bovis BCG[J]. Infect Immun，1996，64：16-22.
[2] Elhay MJ，Oettinger T，Andersen P. Delayed-type hypersensitivity responses to ESAT-6 and MPT64 from Mycobacterium tuberculosis in the guinea pig[J]. Infect Immun，1998，66(7)：3454-3456.
[3] Fan X，Gao Q，FU R.DNA vaccine encoding ESAT-6 enhances the protective efficacy of BCG against Mycobacterium tuberculosis infection in mice[J]. Scand J Immunol，2007，66(5)：523-528.
[4] Munoz S，Hernandez-Pando R，Abraham SN，Enciso JA. Mast cell activation by Mycobacterium tuberculosis:mediator release and role of CD48[J]. J Immunol，2003，170：5590-5596.
[5] Singh G，Singh B，Traikovic V，Sharma P. Mycobacterium tuberculosis 6kDa early secreted antigenic target stimulates activation of J774 macrophages[J]. Immunol Lett，2005，98(2)：180-188.
[6] Cappelli G，Volpe P，Sanduzzi A，Sacchi A,Colizzi V,Mariani F. Human macrophage gamma interferon decreases gene expression but not replication of Mycobacterium tuberculosis: analysis of the host pathogen reciprocal influence on transcription in a comparison of trains H₃₇Rv and CMT97[J]. Infect Immun，2001，69(12)：7262–7270.
[7] Pathak SK， Basu S，Basu KK，Baneriee A，Pathak S,Bhattacharyya A,Kaisho T,Kundu M,Basu J. Direct extracelluar interaction between the early antigen ESAT-6 of Mycobacterium tuberculosis and TLR2 inhibits TLR signaling in macrophages[J]. Nat Immunol，2007，8(6)：610-618.
[8] Yonghong Feng， Xin Yang，Zhonghua Liu，Yaoting Liu，Bo Su，Yuansheng Ding，Lian hua，Qin,Hua Yang，Ruijuan Zheng，Ruiliang Jin. Continuous treatment with recombinant Mycobacterium tuberculosis CFP-10-E SAT-6 protein activated human monocyte while deactivated LPS-stimulated macrophage[J]. Biochem Biophys Res Commun， 2008，365(3)：534-540.
[9] Christoph Holscher，Robert A.A，Berenice Arendse，Najmeeyah Brown，Elmarie Myburgh，Gottfried Alber，Frank Brombacher. A Protective and Agonistic Function of IL-12p40 in mycobacterial infection[J]. J Immunol，2001，167(12)：6957-6966.
[10] Freidag BL，Melton GB，Collins F，Klinman DM，Cheever A，Stobie L，Suen W，Seder RA. CpG oligodeoxynucleotides and interleukin-12 improve the efficacy of Mycobacterium bovis BCG vaccination in mice challenged with M.tuberculosis[J]. Infect Immun，2000，68(5)：2948-2953.
[11] Ha SJ，Park SH，Kim HJ，Kim SC，Kang HJ，Lee EG，Kwon SG，Kim BM，Lee SH，Kim WB，Sung YC，Cho SN. Enhanced immunogenicity and Protective efficacy with the use of interleukin-12-encapsulated microspheres plus AS01B in tuberculosis subunit vaccination[J]. Infect Immun，2006，74(8)：4954-4959.
[12] Cheung BK，Lee DC，Li JC，Lau YL，Lau AS. A role for double-stranded RNA-activated protein kinase PKR in Mycobacterium-induced cytokine expression[J]. J Immunol，2005，175(11)：7218-7225.

[1]	Hu Yifan, Du Boping, Wu Yadong, Zhu Chuanzhi, Zhang Lanyue, Jia Hongyan, Sun Qi, Pan Liping, Zhang Zongde, Li Zihui. Experimental study on the role of Mce4C in the uptake and utilization of cholesterol by Mycobacterium tuberculosis [J]. Chinese Journal of Antituberculosis, 2025, 47(4): 444-453.
[2]	Sheng Jie, Hong Kaifeng, Mierzhati Aisha, Tang Wei, Dilixiati Abulizi. Study on the mechanism of IL-22 and p38 MAPK signaling pathways in inhibiting bone destruction in bone and joint tuberculosis [J]. Chinese Journal of Antituberculosis, 2025, 47(4): 454-459.
[3]	Hao Mingxiao, Mi Jie, Xu Zongyi. Effectiveness of a continuity of care model in patients with tuberculous meningitis [J]. Chinese Journal of Antituberculosis, 2025, 47(4): 477-481.
[4]	Huang Misun, Wu Yaning, Li Guilian, Liu Haican. Research advances of Mycobacterium tuberculosis enrichment technology [J]. Chinese Journal of Antituberculosis, 2025, 47(3): 369-373.
[5]	Zhang Chao, Yu Xia, Huang Hairong, Liu Wei, Liu Tao. Evaluation of the in vitro antimicrobial effects of sevoflurane on Mycobacterium tuberculosis [J]. Chinese Journal of Antituberculosis, 2025, 47(2): 158-163.
[6]	Liu Ruihua, Sarina , Wang Furong. Interaction between lung cancer and tuberculosis in disease development and progression [J]. Chinese Journal of Antituberculosis, 2025, 47(1): 102-111.
[7]	Chen Jifei, Huang Lihua, Luo Lanbo, Sui Wenxian, Pang Yu, Liu Aimei. Evaluation the efficacy of tongue swab-based PCR fluorescence probe method for pulmonary tuberculosis [J]. Chinese Journal of Antituberculosis, 2025, 47(1): 51-60.
[8]	Lu Hailin, Wang Wenfei, Tao Wenhui, Lin Peicong, Chen Xinchun, Deng Guofang, Xie Shuixiang. Oleic acid upregulates the expression of perilipin 2 enhancing macrophage clearance of Mycobacterium tuberculosis [J]. Chinese Journal of Antituberculosis, 2025, 47(1): 72-76.
[9]	Wang Yilin, Wu Xiao, Pang Yu, Li Shanshan. Immunomodulatory effect of orelabrutinib in host macrophages infected with mycobacterium [J]. Chinese Journal of Antituberculosis, 2024, 46(9): 1063-1068.
[10]	Palidanguli Abudureheman, Wang Senlu, Gulina Badeerhan, Wang Le, Zulikatiayi Abudula, Wang Xinqi, Maiwulajiang Yimamu, Wang Xijiang. Distribution of Mycobacterium tuberculosis genotypes in Kashgar region and their association with clinical characteristics of pulmonary tuberculosis patients [J]. Chinese Journal of Antituberculosis, 2024, 46(9): 1077-1082.
[11]	Xu Chunhua, Zhu Shiyu, Hu Yi, Yi Kehua, Song Canlei, Wang Zichun, Wu Yong, Wang Qing, Yang Qianru, Shen Xin. Analysis of screening effect of recombinant Mycobacterium tuberculosis fusion protein in screening Mycobacterium tuberculosis infection in close contacts of pulmonary tuberculosis patients [J]. Chinese Journal of Antituberculosis, 2024, 46(8): 897-902.
[12]	Zhang Lanyue, Wang Yingchao, Liu Weiyi, Shang Xuetian, Jia Hongyan, Zhu Chuanzhi, Zhang Zongde, Pan Liping. Study on the effect of thiol acetyltransferase mshD on the growth and stress response of Mycobacterium tuberculosis [J]. Chinese Journal of Antituberculosis, 2024, 46(8): 935-941.
[13]	Yu Lan, Chen Shuangshuang, Wang Nenhan, Tian Lili, Zhao Yanfeng, Fan Ruifang, Liu Haican, Li Chuanyou, Dai Xiaowei. Consistency between phenotypic resistance to fluoroquinolones and genetic mutations in rifampicin resistant Mycobacterium tuberculosis strains [J]. Chinese Journal of Antituberculosis, 2024, 46(8): 942-950.
[14]	He Xiangrong, Chen Hua, Chen Pinru, Liang Feng, Ren Huili, Zhu Jialou, Hu Jinxing, Tan Yaoju. A case report and literature review of Mycobacterium asiaticum pulmonary disease [J]. Chinese Journal of Antituberculosis, 2024, 46(7): 763-769.
[15]	Zhang Muli, Sun Zhaogang, Cao Tingming, Xie Zhongyao. The value of three proteins in diagnosing Mycobacterium tuberculosis infection [J]. Chinese Journal of Antituberculosis, 2024, 46(7): 808-814.