Stromal Interaction Molecules 1 and 2 Are Key Regulators of Autoreactive T Cell Activation in Murine Autoimmune Central Nervous System Inflammation This information is current as of September 28, 2021. Michael K. Schuhmann, David Stegner, Alejandro Berna-Erro, Stefan Bittner, Attila Braun, Christoph Kleinschnitz, Guido Stoll, Heinz Wiendl, Sven G. Meuth and Bernhard Nieswandt J Immunol 2010; 184:1536-1542; Prepublished online 18 Downloaded from December 2009; doi: 10.4049/jimmunol.0902161 http://www.jimmunol.org/content/184/3/1536 http://www.jimmunol.org/ Supplementary http://www.jimmunol.org/content/suppl/2009/12/18/jimmunol.090216 Material 1.DC1 References This article cites 27 articles, 8 of which you can access for free at: http://www.jimmunol.org/content/184/3/1536.full#ref-list-1 by guest on September 28, 2021 Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists • Fast Publication! 4 weeks from acceptance to publication *average Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2010 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology Stromal Interaction Molecules 1 and 2 Are Key Regulators of Autoreactive T Cell Activation in Murine Autoimmune Central Nervous System Inflammation Michael K. Schuhmann,*,1 David Stegner,†,1 Alejandro Berna-Erro,† Stefan Bittner,* Attila Braun,† Christoph Kleinschnitz,* Guido Stoll,* Heinz Wiendl,* Sven G. Meuth,* and Bernhard Nieswandt† Calcium (Ca2+) signaling in T lymphocytes is essential for a variety of functions, including the regulation of differentiation, gene transcription, and effector functions. A major Ca2+ entry pathway in nonexcitable cells, including T cells, is store-operated Ca2+ entry (SOCE), wherein depletion of intracellular Ca2+ stores upon receptor stimulation causes subsequent influx of extracellular Ca2+ across the plasma membrane. Stromal interaction molecule (STIM) 1 is the Ca2+ sensor in the endoplasmic reticulum, which Downloaded from controls this process, whereas the other STIM isoform, STIM2, coregulates SOCE. Although the contribution of STIM molecules and SOCE to T lymphocyte function is well studied in vitro, their significance for immune processes in vivo has remained largely elusive. In this study, we studied T cell function in mice lacking STIM1 or STIM2 in a model of myelin-oligodendrocyte glycoprotein (MOG35–55)-induced experimental autoimmune encephalomyelitis (EAE). We found that STIM1 deficiency significantly impaired the generation of neuroantigen-specific T cell responses in vivo with reduced Th1/Th17 responses, resulting in complete protection from EAE. Mice lacking STIM2 developed EAE, but the disease course was ameliorated. This was associated with a reduced clinical http://www.jimmunol.org/ peak of disease. Deficiency of STIM2 was associated with an overall reduced proliferative capacity of lymphocytes and a reduction of IFN-g/IL-17 production by neuroantigen-specific T cells. Neither STIM1 nor STIM2 deficiency altered the phenotype or function of APCs. These findings reveal a crucial role of STIM-dependent pathways for T cell function and activation under autoimmune inflammatory conditions, establishing them as attractive new molecular therapeutic targets for the treatment of inflammatory and autoimmune disorders. The Journal of Immunology, 2010, 184: 1536–1542. cells orchestrate inflammation in multiple sclerosis, an diacylglycerol and 1,4,5-inositol trisphosphate, which triggers Ca2+ inflammatory disorder of the CNS resulting in demye- release from the endoplasmic reticulum through 1,4,5-inositol by guest on September 28, 2021 lination, oligodendrocyte loss, and axonal damage (1, 2). trisphosphate receptor channels and leads to a transient elevation T 2+ 2+ Calcium signals control proliferation, differentiation, apoptosis, of the cytosolic Ca concentration (7). This Ca store depletion is and a variety of transcriptional programs and effector functions of followed by Ca2+ entry through specialized Ca2+ release-activated T cells (3–5). Conversely, disturbed Ca2+ signaling and dysregu- calcium channels in the plasma membrane, resulting in longer- lation of the intracellular Ca2+ network in T lymphocytes have lasting (∼1 h) elevated cytosolic Ca2+ concentration, which is been associated with inflammatory and autoimmune disorders, mandatory for transcription-dependent steps of T cell activation including multiple sclerosis (6). (8, 9). This so-called store-operated calcium entry (SOCE) occurs Signaling cascades after TCR stimulation involve phospholipase through the four-transmembrane channel, Orai1, (6, 10, 11) and is C-mediated cleavage of phosphotidylinositol 4,5-bisphosphate into regulated by the endoplasmic reticulum-resident calcium sensor molecule stromal interaction molecule (STIM) 1, which connects *Department of Neurology and †Rudolf Virchow Center, Deutsche Forschungsge- store depletion to the opening of the channel (12). The closely meinschaft Research Center for Experimental Biomedicine, University of Wu¨rzburg, related STIM2 has been shown to regulate basal cytosolic and Wu¨rzburg, Germany store Ca2+ concentrations in different cell types (13). The recent 1 M.K.S. and D.S. contributed equally to this work. generation of mice lacking STIM molecules in T cells confirmed Received for publication July 7, 2009. Accepted for publication November 20, 2009. the essential function of STIM1 for SOCE (12, 14) and also re- This work was supported by the Rudolf Virchow Center and the Deutsche For- vealed the contribution of STIM2 to this process (15). The sig- schungsgemeinschaft (Sonderforschungsbereiche Grants 487 and 688 to B.N. and nificance of SOCE for T cell function in vivo, however, has not Grant 581 to S.G.M.). D.S. was supported by a grant of the German Excellence Initiative to the Graduate School of Life Sciences, University of Wu¨rzburg. been definitively addressed to date despite the recent description Address correspondence and reprint requests to Dr. Sven G. Meuth, Department of of human immunodeficiency syndromes associated with defi- Neurology, Josef-Schneider-Straße 11, 97080 Wu¨rzburg, Germany, or Dr. Bernhard ciencies in STIM1 function (16). A first study has revealed that Nieswandt, Rudolf-Virchow Zentrum, Josef-Schneider-Straße 2, 97080 Wu¨rzburg STIM1-deficient T cells display defects in homeostatic T cell Germany. E-mail addresses: [email protected] and bernhard.nieswandt @virchow.uni-wuerzburg.de proliferation but, despite abolished SOCE, are able to support The online version of this paper contains supplemental material. humoral immune responses after vaccination and can induce acute Abbreviations used in this paper: aGVHD, acute graft-versus-host disease; bd, below graft-versus-host disease following adoptive transfer into alloge- detection level; BM, bone marrow; DC, dendritic cell; EAE, experimental autoimmune neic hosts (12). In contrast, the significance of STIM-dependent encephalomyelitis; LFB, Luxol fast blue; MOG, myelin-oligodendrocyte glycoprotein; SOCE for inflammatory reactions in vivo has remained elusive. SOCE, store-operated Ca2+ entry; STIM, stromal interaction molecule; wt, wild-type. Inthisstudy,weinvestigatethecontributionofSTIM1andSTIM2in Copyright Ó 2010 by The American Association of Immunologists, Inc. 0022-1767/10/$16.00 a model of autoimmune CNS inflammation, myelin oligodendrocyte www.jimmunol.org/cgi/doi/10.4049/jimmunol.0902161 The Journal of Immunology 1537 glycoprotein (MOG35–55) peptide-induced experimental autoimmune rat anti-mouse CD8a-PE (no. 553033), rat anti-mouse CD44-FITC (no. encephalomyelitis (EAE). We show that STIM1 deficiency abrogates 553133), rat anti-mouse CD62L-APC (no. 553152), rat anti-mouse CD11b- disease development, whereas STIM2 deficiency significantly delays PerCP (no. 350993), hamster anti-mouse CD11c-APC (no. 550261), mouse anti-mouse MCHII-FITC (no. MCA1501F; Serotec, Du¨sseldorf, Germany), onset and attenuates the clinical course of MOG EAE. These findings rat anti-mouse CD86-FITC (no. 553691), hamster anti-mouse CD80-FITC provide a strong rationale for considering STIM molecules as po- (no. 553768), and hamster anti-mouse CD40-FITC (no. 553723). 2 2 tential new pharmacological targets in T cell-mediated disorders. For functional analysis, dentritic cells (DCs) from wt or Stim2 / mice were prepared from BM cells flushed from femur and tibia bones as de- scribed before (19). BM-derived DCs were harvested on day 8, and 2 3 106 Materials and Methods cells in 1 ml splenocyte complete medium were incubated with 50 mg/ml Mice 5 MOG35–55 for 6 h. Thereafter MOG35–55-loaded DCs (1 3 10 ) were co- + 3 5 The generation of Stim12/2 bone marrow (BM) chimeric mice has been cultured with MACS isolated CD4 T cells (5 10 ) (Miltenyi Biotec, described previously (17). Stim12/2 BM chimeric mice or equally treated Bergisch Gladbach, Germany) for 3 d, and supernatants were assessed for wild-type (wt) mice were used 11–20 wk after transplantation.