PMID- 31816492 OWN - NLM STAT- MEDLINE DCOM- 20200409 LR - 20200409 IS - 1872-9142 (Electronic) IS - 0161-5890 (Linking) VI - 117 DP - 2020 Jan TI - Early secreted antigenic target of 6-kDa of Mycobacterium tuberculosis induces transition of macrophages into epithelioid macrophages by downregulating iNOS / NO-mediated H3K27 trimethylation in macrophages. PG - 189-200 LID - S0161-5890(19)30437-7 [pii] LID - 10.1016/j.molimm.2019.11.013 [doi] AB - BACKGROUND: Tuberculosis (TB) is a chronic infectious disease caused by Mycobacterium tuberculosis (Mtb). Granuloma is a pathological feature of tuberculosis and is a tight immune cell aggregation caused by Mtb. The main constituent cells are macrophages and their derivative cells including epithelioid macrophages. However, the molecular mechanism of the transition has not been reported. The purpose of this study was to investigate whether early secreted antigenic target of 6-kDa (ESAT6) can induce the transition of bone marrow-derived macrophages (BMDMs) into epithelioid macrophages and its possible molecular mechanism. METHODS: The recombinant ESAT6 protein was obtained from E.coli carrying esat6 gene after isopropyl beta-d-thiogalactopyranoside (IPTG) induction. BMDMs were isolated from bone marrow of mice hind legs. Cells viability was detected by Cell Counting Kit 8 (CCK8) assays. The expression levels of mRNA and proteins were detected by qPCR and Western blot, or evaluated by flow cytometry. The expression level of nitric oxide (NO) was measured with a nitric oxide indicator. RESULTS: ESAT6 could significantly induce mRNA and protein expression levels of a group of epithelioid macrophages marker molecules (EMMMs), including E-cadherin, junction plakoglobin, ZO1, desmoplakin, desmoglein3 and catenin porteins, in BMDMs. These events could be abrogated in macrophage from TLR2 deficiency mice. ESAT6 could also markedly induce iNOS/NO production that could significantly inhibit trimethylation of H3K27 in the cells. ESAT6-induced expressions of epithelioid macrophages marker molecules were significantly inhibited in the presence of H3K27 histone demethylase inhibitor GSK J1. Furthermore, ROS scavenging agent N,N'-Dimethylthiourea (DMTU) could markedly inhibit the transition induced by ESAT6 in macrophages. CONCLUSION: This study demonstrates that ESAT6 bound with TLR2 can activate iNOS/NO and ROS signalings to reduce the trimethylation of H3K27 resulting in the increment of EMMMs expression that is beneficial to the transition of macrophages into epithelioid macrophages. However, hypoxia can inhibit this transition event. This study has provided new evidence of pathogenesis of granuloma caused by Mtb and also proposed new ideas for the treatment of TB. CI - Copyright (c) 2019 Elsevier Ltd. All rights reserved. FAU - Lin, Jiahui AU - Lin J AD - Shanghai Key Lab of Tuberculosis, Shanghai Pulmonary Hospital, School of Medicine, Tongji University, 507 Zhengmin Road, Shanghai, 200433, China; Department of Microbiology and Immunology, School of Medicine, Tongji University, 1239 Siping Road, Shanghai, 200092, China. FAU - Jiang, Yuyin AU - Jiang Y AD - Shanghai Key Lab of Tuberculosis, Shanghai Pulmonary Hospital, School of Medicine, Tongji University, 507 Zhengmin Road, Shanghai, 200433, China; Department of Microbiology and Immunology, School of Medicine, Tongji University, 1239 Siping Road, Shanghai, 200092, China. FAU - Liu, Dan AU - Liu D AD - Shanghai Key Lab of Tuberculosis, Shanghai Pulmonary Hospital, School of Medicine, Tongji University, 507 Zhengmin Road, Shanghai, 200433, China; Department of Microbiology and Immunology, School of Medicine, Tongji University, 1239 Siping Road, Shanghai, 200092, China. FAU - Dai, Xueting AU - Dai X AD - Department of Microbiology and Immunology, School of Medicine, Tongji University, 1239 Siping Road, Shanghai, 200092, China. FAU - Wang, Min AU - Wang M AD - Shanghai Key Lab of Tuberculosis, Shanghai Pulmonary Hospital, School of Medicine, Tongji University, 507 Zhengmin Road, Shanghai, 200433, China; Department of Microbiology and Immunology, School of Medicine, Tongji University, 1239 Siping Road, Shanghai, 200092, China. FAU - Dai, Yalei AU - Dai Y AD - Shanghai Key Lab of Tuberculosis, Shanghai Pulmonary Hospital, School of Medicine, Tongji University, 507 Zhengmin Road, Shanghai, 200433, China; Department of Microbiology and Immunology, School of Medicine, Tongji University, 1239 Siping Road, Shanghai, 200092, China. Electronic address: daiyl@tongji.edu.cn. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20191206 PL - England TA - Mol Immunol JT - Molecular immunology JID - 7905289 RN - 0 (Antigens, Bacterial) RN - 0 (Bacterial Proteins) RN - 0 (ESAT-6 protein, Mycobacterium tuberculosis) RN - 0 (Histones) RN - 31C4KY9ESH (Nitric Oxide) RN - EC 1.14.13.39 (Nitric Oxide Synthase Type II) SB - IM MH - Animals MH - Antigens, Bacterial/*metabolism MH - Bacterial Proteins/*metabolism MH - Cell Transdifferentiation/*physiology MH - DNA Methylation/physiology MH - Down-Regulation MH - Granuloma/metabolism/microbiology/pathology MH - Histones MH - Macrophages/*metabolism/pathology MH - Mice MH - Mycobacterium tuberculosis MH - Nitric Oxide/metabolism MH - Nitric Oxide Synthase Type II/metabolism MH - Signal Transduction/*physiology MH - Tuberculosis/*metabolism/pathology OTO - NOTNLM OT - ESAT6 OT - Granuloma OT - H3K27me3 OT - Macrophage OT - NO OT - TLR2 EDAT- 2019/12/10 06:00 MHDA- 2020/04/10 06:00 CRDT- 2019/12/10 06:00 PHST- 2019/06/14 00:00 [received] PHST- 2019/10/31 00:00 [revised] PHST- 2019/11/30 00:00 [accepted] PHST- 2019/12/10 06:00 [pubmed] PHST- 2020/04/10 06:00 [medline] PHST- 2019/12/10 06:00 [entrez] AID - S0161-5890(19)30437-7 [pii] AID - 10.1016/j.molimm.2019.11.013 [doi] PST - ppublish SO - Mol Immunol. 2020 Jan;117:189-200. doi: 10.1016/j.molimm.2019.11.013. Epub 2019 Dec 6.