Clinical Epigenetics | |
Hypomethylation and overexpression of ITGAL (CD11a) in CD4+ T cells in systemic sclerosis | |
Rong Xiao3 QianJin Lu4 Ming Zhao4 JiuCun Wang1 YaPing Li3 Takuro Kanekura5 Qing Wang3 YangFan Xiao3 Ye Shu2 YaoYao Wang6 | |
[1] Ministry of Education (MOE) Key Laboratory of Contemporary Anthropology and State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University, 220 Handan Road, 200433 Shanghai, China;Department of Dermatology, Hunan Children’s Hospital, 86 Zi-Yuan Road, Changsha 410007, China;Department of Dermatology, Second Xiangya Hospital, Central South University, 139 Ren-Min Road, Changsha 410011, China;Hunan Key Laboratory of Medical Epigenomics, 139 Ren-Min Road, Changsha 410011, China;Department of Dermatology, Kagoshima University Graduate School of Medical and Dental Sciences, 8-35-1 Sakuragaoka, Kagoshima 890-8520, Japan;Department of Dermatology, Sir Run Run Shaw Hospital, Zhejiang University, 3 East Qingchun Road, Hangzhou 310016, China | |
关键词: Systemic sclerosis; DNA methylation; COL1A2; CD4+ T cells; CD11a; | |
Others : 1092790 DOI : 10.1186/1868-7083-6-25 |
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received in 2014-08-12, accepted in 2014-10-24, 发布年份 2014 | |
【 摘 要 】
Background
The pathogenesis and etiology of systemic sclerosis (SSc) are complex and poorly understood. To date, several studies have demonstrated that the activation of the immune system undoubtedly plays a pivotal role in SSc pathogenesis. Activated immune effector T cells contribute to the release of various pro-inflammatory cytokines and drive the SSc-specific autoantibody responses. This, and a profibrotic environment, are all-important components of abnormal active immune responses that can lead to pathological disorders of SSc. CD11a is essential to inflammatory and immune responses, regulating adhesive and co-stimulatory interactions between CD4+ T cells and other cells. Although CD11a is overexpressed in SSc patients, the mechanisms leading to this overexpression and its consequences remain unclear. DNA methylation, a main epigenetic modification, plays an important role in the regulation of gene expression and is involved in the pathogenesis of autoimmune diseases. This work aims to investigate the effect of DNA demethylation on CD11a expression in SSc CD4+ T cells and to determine its functional significance. CD11a expression was measured using RT-PCR and flow cytometry. Bisulfite sequencing was used to determine the methylation status of the CD11a regulatory region. CD4+ T cells were co-cultured with antigen-presenting cells, B cells, or fibroblasts with and without anti-CD11a, and proliferation of CD4+ T cells, IgG production by B cells, and expression levels of COL1A2 mRNA by fibroblasts were evaluated.
Results
Elevated CD11a expression levels were observed in CD4+ T cells from SSc patients; these levels were found to be positively correlated with disease activity. The methylation levels of the CD11a regulatory sequences were lower in SSc patients than in controls and inversely correlated with CD11a mRNA expression. Treatment of CD4+ T cells with 5-azacytidine (5-azaC) decreased CD11a promoter methylation and caused CD11a overexpression. SSc CD4+ T cells and 5-azaC-treated CD4+ T cells showed increased proliferation of CD4+ T cells, increased production of IgG by co-cultured B cells, and induced expression of COL1A2 mRNA by co-cultured fibroblasts. These stimulatory effects were abrogated by anti-CD11a.
Conclusions
Demethylation of CD11a regulatory elements and subsequent CD11a overexpression in CD4+ T cells may mediate immunological abnormalities and fibrotic processes in SSc.
【 授权许可】
2014 Wang et al.; licensee BioMed Central Ltd.
【 预 览 】
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【 参考文献 】
- [1]Chizzolini C, Brembilla NC, Montanari E, Truchetet ME: Fibrosis and immune dysregulation in systemic sclerosis. Autoimmun Rev 2011, 10:276-281.
- [2]Cutolo M, Sulli A, Smith V: Assessing microvascular changes in systemic sclerosis diagnosis and management. Nat Rev Rheumatol 2010, 6:578-587.
- [3]Postlethwaite AE, Shigemitsu H, Kanangat S: Cellular origins of fibroblasts: possible implications for organ fibrosis in systemic sclerosis. Curr Opin Rheumatol 2004, 16:733-738.
- [4]Gu YS, Kong J, Cheema GS, Keen CL, Wick G, Gershwin ME: The immunobiology of systemic sclerosis. Semin Arthritis Rheum 2008, 38:132-160.
- [5]O’Reilly S, Hügle T, van Laar JM: T cells in systemic sclerosis: a reappraisal. Rheumatology (Oxford) 2012, 51:1540-1549.
- [6]Chizzolini C: T cells, B cells, and polarized immune response in the pathogenesis of fibrosis and systemic sclerosis. Curr Opin Rheumatol 2008, 20:707-712.
- [7]Shimaoka M, Springer TA: Therapeutic antagonists and conformational regulation of integrin function. Nat Rev Drug Discov 2003, 2:703-716.
- [8]Yusuf-Makagiansar H, Anderson ME, Yakovleva TV, Murray JS, Siahaan TJ: Inhibition of LFA-1/ICAM-1 and VLA-4/VCAM-1 as a therapeutic approach to inflammation and autoimmune diseases. Med Res Rev 2002, 22:146-167.
- [9]Stummvoll GH, Aringer M, Grisar J, Steiner CW, Smolen JS, Knobler R, Graninger WB: Increased transendothelial migration of scleroderma lymphocytes. Ann Rheum Dis 2004, 63:569-574.
- [10]Manetti M, Neumann E, Müller A, Schmeiser T, Saar P, Milia AF, Endlicher E, Roeb E, Messerini L, Matucci-Cerinic M, Ibba-Manneschi L, Müller-Ladner U: Endothelial/lymphocyte activation leads to prominent CD4+ T cell infiltration in the gastric mucosa of patients with systemic sclerosis. Arthritis Rheum 2008, 58:2866-2873.
- [11]Gruschwitz MS, Hornstein OP, von Den DP: Correlation of soluble adhesion molecules in the peripheral blood of scleroderma patients with their in situ expression and with disease activity. Arthritis Rheum 1995, 38:184-189.
- [12]Sawaya HH, de Souza RB, Carrasco S, Goldenstein-Schainberg C: Altered adhesion molecules expression on peripheral blood mononuclear cells from patients with systemic sclerosis and clinical correlations. Clin Rheumatol 2009, 28:847-851.
- [13]Rabquer BJ, Hou Y, Del Galdo F, Kenneth Haines G III, Gerber ML, Jimenez SA, Seibold JR, Koch AE: The proadhesive phenotype of systemic sclerosis skin promotes myeloid cell adhesion via ICAM-1 and VCAM-1. Rheumatology (Oxford) 2009, 48:734-740.
- [14]Yoshizaki A, Yanaba K, Iwata Y, Komura K, Ogawa A, Akiyama Y, Muroi E, Hara T, Ogawa F, Takenaka M, Shimizu K, Hasegawa M, Fujimoto M, Tedder TF, Sato S: Cell adhesion molecules regulate fibrotic process via Th1/Th2/Th17 cell balance in a bleomycin-induced scleroderma model. J Immunol 2010, 185:2502-2515.
- [15]Luo Y, Wang Y, Wang Q, Xiao R, Lu Q: Systemic sclerosis: genetics and epigenetics. J Autoimmun 2013, 41:161-167.
- [16]Feghali-Bostwick C, Medsger TA Jr, Wright TM: Analysis of systemic sclerosis in twins reveals low concordance for disease and high concordance for the presence of antinuclear antibodies. Arthritis Rheum 2003, 48:1956-1963.
- [17]Broen JC, Radstake TR, Rossato M: The role of genetics and epigenetics in the pathogenesis of systemic sclerosis. Nat Rev Rheumatol 2014, 10(11):671-681.
- [18]De Santis M, Selmi C: The therapeutic potential of epigenetics in autoimmune diseases. Clin Rev Allergy Immunol 2012, 42:92-101.
- [19]Meda F, Folci M, Baccarelli A, Selmi C: The epigenetics of autoimmunity. Cell Mol Immunol 2011, 8:226-236.
- [20]Selmi C, Feghali-Bostwick CA, Lleo A, Lombardi SA, De Santis M, Cavaciocchi F, Zammataro L, Mitchell MM, Lasalle JM, Medsger T Jr, Gershwin ME: X chromosome gene methylation in peripheral lymphocytes from monozygotic twins discordant for scleroderma. Clin Exp Immunol 2012, 169:253-262.
- [21]Lu Q, Ray D, Gutsch D, Richardson B: Effect of DNA methylation and chromatin structure on ITGAL expression. Blood 2002, 99:4503-4508.
- [22]Richardson BC, Powers D, Hooper F, Yung RL, O’Rourke K: Lymphocyte function-associated antigen 1 overexpression and T cell autoreactivity. Arthritis Rheum 1994, 37:1363-1372.
- [23]Yung R, Powers D, Johnson K, Amento E, Carr D, Laing T, Yang J, Chang S, Hemati N, Richardson B: Mechanisms of drug-induced lupus. II. T cells overexpressing lymphocyte function-associated antigen 1 become autoreactive and cause a lupus-like disease in syngeneic mice. J Clin Invest 1996, 97:2866-2871.
- [24]Lu Q: The critical importance of epigenetics in autoimmunity. J Autoimmun 2013, 41:1-5.
- [25]Kaplan MJ, Lu Q, Wu A, Attwood J, Richardson B: Demethylation of promoter regulatory elements contributes to perforin overexpression in CD4+ lupus T cells. J Immunol 2004, 172:3652-3661.
- [26]Lei W, Luo Y, Lei W, Luo Y, Yan K, Zhao S, Li Y, Qiu X, Zhou Y, Long H, Zhao M, Liang Y, Su Y, Lu Q: Abnormal DNA methylation in CD4+ T cells from patients with systemic lupus erythematosus, systemic sclerosis, and dermatomyositis. Scand J Rheumatol 2009, 38:369-374.
- [27]Lian X, Xiao R, Hu X, Kanekura T, Jiang H, Li Y, Wang Y, Yang Y, Zhao M, Lu Q: DNA demethylation of CD40l in CD4+ T cells from women with systemic sclerosis: a possible explanation for female susceptibility. Arthritis Rheum 2012, 64:2338-2345.
- [28]Jiang H, Xiao R, Lian X, Kanekura T, Luo Y, Yin Y, Zhang G, Yang Y, Wang Y, Zhao M, Lu Q: Demethylation of TNFSF7 contributes to CD70 overexpression in CD4+ T cells from patients with systemic sclerosis. Clin Immunol 2012, 143:39-44.
- [29]Lu Q, Kaplan M, Ray D, Ray D, Zacharek S, Gutsch D, Richardson B: Demethylation of ITGAL (CD11a) regulatory sequences in systemic lupus erythematosus. Arthritis Rheum 2002, 46:1282-1291.
- [30]Shelley CS, Farokhzad OC, Arnaout MA: Identification of cell-specific and developmentally regulated nuclear factors that direct myeloid and lymphoid expression of the CD11a gene. Proc Natl Acad Sci U S A 1993, 90:5364-5368.
- [31]Puig-Kröger A, Sanchez-Elsner T, Ruiz N, Andreu EJ, Prosper F, Jensen UB, Gil J, Erickson P, Drabkin H, Groner Y, Corbi AL: RUNX/AML and C/EBP factors regulate CD11a integrin expression in myeloid cells through overlapping regulatory elements. Blood 2003, 102:3252-3261.
- [32]Domínguez-Soto A, Relloso M, Vega MA, Corbí AL, Puig-Kröger A: RUNX3 regulates the activity of the CD11a and CD49d integrin gene promoters. Immunobiology 2005, 210:133-139.
- [33]Cornwell RD, Gollahon KA, Hickstein DD: Description of the leukocyte function-associated antigen 1 (LFA-1 or CD11a) promoter. Proc Natl Acad Sci U S A 1993, 90:4221-4225.
- [34]Lai AY, Mav D, Shah R, Grimm SA, Phadke D, Hatzi K, Melnick A, Geigerman C, Sobol SE, Jaye DL, Wade PA: DNA methylation profiling in human B cells reveals immune regulatory elements and epigenetic plasticity at Alu elements during B cell activation. Genome Res 2013, 23:2030-2041.
- [35]Jintaridth P, Tungtrongchitr R, Preutthipan S, Mutirangura A: Hypomethylation of Alu elements in post-menopausal women with osteoporosis. PLoS One 2013, 8:e70386.
- [36]Mehra S, Walker J, Patterson K, Fritzler MJ: Autoantibodies in systemic sclerosis. Autoimmun Rev 2013, 12:340-354.
- [37]Holland J, Owens T: Signaling through intercellular adhesion molecule 1 (ICAM-1) in a B cell lymphoma line: the activation of Lyn tyrosine kinase and the mitogen-activated protein kinase pathway. J Biol Chem 1997, 272:9108-9112.
- [38]Tohma S, Hirohata S, Lipsky PE: The role of CD11a/CD18-CD54 interactions in human T cell-dependent B cell activation. J Immunol 1991, 146:492-499.
- [39]Cliff JM, Klaus GG: A method for investigating the role of homotypic adhesion in lymphocyte activation. J Immunol Methods 2000, 246:51-59.
- [40]Carrasco YR, Fleire SJ, Cameron T, Dustin ML, Batista FD: LFA-1/ICAM-1 interaction lowers the threshold of B cell activation by facilitating B cell adhesion and synapse formation. Immunity 2004, 20:589-599.
- [41]López-Hoyos M, Revilladagger C, Conde C, Del Campo EG, González A, Merino J: Different roles for LFA-1 and VLA-4 integrins in T-B-cell interactions in vivo. Immunology 1999, 97:438-446.
- [42]Arana E, Vehlow A, Harwood NE, Vigorito E, Henderson R, Turner M, Tybulewicz VL, Batista FD: Activation of the small GTPase Rac2 via the B cell receptor regulates B cell adhesion and immunological-synapse formation. Immunity 2008, 28:88-99.
- [43]Lonzetti LS, Joyal F, Raynauld JP, Roussin A, Goulet JR, Rich E, Choquette D, Raymond Y, Senécal JL: Updating the American College of Rheumatology preliminary classification criteria for systemic sclerosis: addition of severe nailfoldcapillaroscopy abnormalities markedly increases the sensitivity for limited scleroderma. Arthritis Rheum 2001, 44:735-736.
- [44]Shima Y, Kuwahara Y, Murota H, Kitaba S, Kawai M, Hirano T, Arimitsu J, Narazaki M, Hagihara K, Ogata A, Katayama I, Kawase I, Kishimoto T, Tanaka T: The skin of patients with systemic sclerosis softened during the treatment with anti-IL-6 receptor antibody tocilizuma. Rheumatology (Oxford) 2010, 49:2408-2412.
- [45]Hudson M, Steele R, Baron M, Canadian Scleroderma Research Group (CSRG): Update on indices of disease activity in systemic sclerosis. Semin Arthritis Rheum 2007, 37:93-98.
- [46]Zhou Y, Yuan J, Pan Y, Fei Y, Qiu X, Hu N, Luo Y, Lei W, Li Y, Long H, Sawalha AH, Richardson B, Lu Q: T cell CD40LG gene expression and the production of IgG by autologous B cells in systemic lupus erythematosus. Clin Immunol 2009, 132:362-370.
- [47]Xiao R, Yoshida N, Higashi Y, Lu QJ, Fukushige T, Kanzaki T, Kanekura T: Retinoic acids exhibit anti-fibrotic activity through the inhibition of 5-lipoxygenase expression in scleroderma fibroblasts. J Dermatol 2011, 38:345-353.
- [48]Ueda-Hayakawa I, Hasegawa M, Hamaguchi Y, Takehara K, Fujimoto M: Circulating γ/δ T cells in systemic sclerosis exhibit activated phenotype and enhance gene expression of proalpha2(I) collagen of fibroblasts. J Dermatol Sci 2013, 69:54-60.