MBD2 regulates T H 17 differentiation and experimental autoimmune encephalomyelitis by controlling the homeostasis of T-bet/Hlx axis Jixin Zhong a, 1 , Qilin Yu a, 1 , Ping Yang a, 1 , Xiaoquan Rao b , Long He a , Jing Fang a , Yaqin Tu a , Zhijun Zhang a , Qiaohong Lai a , Shu Zhang a , Michal Kuczma b , Piatr Kraj b , Jun-Fa Xu c , Feili Gong d , Jianfeng Zhou e , Li Wen f , Decio L. Eizirik g , Jie Du h , Wei Wang i, ** , Cong-Yi Wang a, * a The Center for Biomedical Research, Tongji Hospital, Huazhong University of Science and Technology,1095 Jiefang Ave., Wuhan 430030, China b The Center for Biotechnology and Genomic Medicine, Georgia Regents University,1120 15th Street, Augusta, GA 30912, USA c Department of Clinical Immunology, Institute of Laboratory Medicine, Guangdong Medical College, Dongguan 523808, China d Department of Immunology, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China e Department of Hematology, Tongji Hospital, Huazhong University of Science and Technology,1095 Jiefang Ave., Wuhan 430030, China f Section of Endocrinology, Department of Internal Medicine, Yale University School of Medicine, New Haven, CT 06520, USA g Laboratory of Experimental Medicine, Universite Libre de Bruxelles, Route de Lennik 808, CP 618, B-1070 Brussels, Belgium h Beijing Anzhen Hospital, Capital Medical University, Beijing Institute of Heart, Lung and Blood Vessel Diseases, The Key Laboratory of Remodeling-Related Cardiovascular Diseases, Ministry of Education, Beijing, China i Department of Neurology, Tongji Hospital, Huazhong University of Science and Technology,1095 Jiefang Ave., Wuhan 430030, China article info Article history: Received 21 March 2014 Received in revised form 17 May 2014 Accepted 23 May 2014 Available online 14 June 2014 Keywords: EAE Epigenetic Methylation MBD2 T H 17 T-bet abstract Unlike genetic alterations, epigenetic modifications are reversible and amenable to pharmacological interventions, which make them appealing targets for clinical therapy. However, little is known about epigenetic regulation in experimental autoimmune encephalomyelitis (EAE). Here we demonstrated that methyl-CpG-binding domain protein 2 (MBD2), an epigenetic regulator, controls autoimmunity and EAE through T-bet/Hlx. Tbx21 and Hlx underwent a DNA methylation turnover upon polarizations and a unique methylation pattern was essential for T H 17 development. Loss of Mbd2 resulted in a defect for reading the information encoded by this methylation turnover, which disrupted the homeostasis of T- bet/Hlx axis and suppressed T H 17 differentiation. DNA demethylation induced similar effect on helper T cell differentiation. Therefore, Mbd2 / mice were completely protected from EAE. Pathogenic spleno- cytes isolated from wild-type mice challenged with MOG35-55 could adoptively transfer disease to Mbd2 / mice. In addition, Mbd2 / mice reconstituted with unstimulated wild-type splenocytes developed EAE as wild-type mice did. These data would provide novel insights into epigenetic regulation of EAE. © 2014 Elsevier Ltd. All rights reserved. 1. Introduction Methylation of CpG DNA is the only genetically programmed DNA modification in mammals [1], which encodes inheritable in- formation to control a relatively stable or homeostatic profile of gene expression essential for biological processes [2]. As a result, DNA methylation has been demonstrated to act as a common strategy encoding information superimposed on DNA sequences for the regulation of immune response [3]. Of important note, unlike its nuclear genome counterpart, even identical monozygotic twins are generally associated with disparities in terms of DNA methyl- ome, which are thought to be caused by the differences of envi- ronmental exposures [4]. Indeed, changes on DNA methylation Abbreviations: BMDCs, bone marrow-derived DCs; CFU, colony-forming units; ChIP, chromatin immunoprecipitation; CLP, cecal ligation and puncture; DCs, den- dritic cells; EAE, experimental autoimmune encephalomyelitis; EMSA, electropho- retic mobility shift assay; Hlx, H 2.0-like homeobox; MBD2, methyl-CpG binding domain protein 2; MOG35-55, myelin oligodendrocyte glycoprotein peptide 35-55; Treg, regulatory T cell. * Corresponding author. Tel.: þ86 27 8366 3485; fax: þ86 27 8366 3486. ** Corresponding author. E-mail addresses: wwang@tjh.tjmu.edu.cn (W. Wang), wangcy@tjh.tjmu.edu.cn (C.-Y. Wang). 1 These authors contributed equally to this work. Contents lists available at ScienceDirect Journal of Autoimmunity journal homepage: www.elsevier.com/locate/jautimm http://dx.doi.org/10.1016/j.jaut.2014.05.006 0896-8411/© 2014 Elsevier Ltd. All rights reserved. Journal of Autoimmunity 53 (2014) 95e104