Endogenous CD83 Expression in CD4+ Conventional T Cells Controls Inflammatory Immune Responses This information is current as Katarina Liedtke, Christina Alter, Anne Günther, Nadine of September 28, 2021. Hövelmeyer, Robert Klopfleisch, Ronald Naumann, F. Thomas Wunderlich, Jan Buer, Astrid M. Westendorf and Wiebke Hansen J Immunol published online 27 April 2020 http://www.jimmunol.org/content/early/2020/04/24/jimmun Downloaded from ol.2000042 Supplementary http://www.jimmunol.org/content/suppl/2020/04/25/jimmunol.200004 Material 2.DCSupplemental http://www.jimmunol.org/ Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! 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Published April 27, 2020, doi:10.4049/jimmunol.2000042 The Journal of Immunology Endogenous CD83 Expression in CD4+ Conventional T Cells Controls Inflammatory Immune Responses Katarina Liedtke,* Christina Alter,* Anne Gu¨nther,* Nadine Ho¨velmeyer,† Robert Klopfleisch,‡ Ronald Naumann,x F. Thomas Wunderlich,{,‖ Jan Buer,* Astrid M. Westendorf,* and Wiebke Hansen* The glycoprotein CD83 is known to be expressed by different immune cells including activated CD4+Foxp3+ regulatory T cells (Tregs) and CD4+Foxp32 conventional T cells. However, the physiological function of endogenous CD83 in CD4+ T cell subsets is still unclear. In this study, we have generated a new CD83flox mouse line on BALB/c background, allowing for specific ablation of CD83 in T cells upon breeding with CD4-cre mice. Tregs from CD83flox/flox/CD4-cretg/wt mice had similar suppressive activity as Tregs from CD83flox/flox/CD4-crewt/wt wild-type littermates, suggesting that endogenous CD83 expression is dispensable for the inhibitory capacity of Tregs. However, CD83-deficient CD4+ conventional T cells showed elevated proliferation and IFN-g Downloaded from secretion as well as an enhanced capacity to differentiate into Th1 cells and Th17 cells upon stimulation in vitro. T cell–specific ablation of CD83 expression resulted in aggravated contact hypersensitivity reaction accompanied by enhanced CD4+ T cell activation. Moreover, adoptive transfer of CD4+CD45RBhigh T cells from CD83flox/flox/CD4-cretg/wt mice into Rag2-deficient mice elicited more severe colitis associated with increased serum concentrations of IL-12 and elevated CD40 expression on CD11c+ dendritic cells (DCs). Strikingly, DCs from BALB/c mice cocultured with CD83-deficient CD4+ conventional T cells showed enhanced CD40 expression and IL-12 secretion compared with DCs cocultured with CD4+ conventional T cells from http://www.jimmunol.org/ CD83flox/flox/CD4-crewt/wt wild-type mice. In summary, these results indicate that endogenous CD83 expression in CD4+ conventional T cells plays a crucial role in controlling CD4+ T cell responses, at least in part, by regulating the activity of CD11c+ DCs. The Journal of Immunology, 2020, 204: 000–000. he glycoprotein CD83 is a type 1 transmembrane protein In addition to the full-length membrane-bound form of CD83, a and member of the Ig superfamily (1). Genetic and soluble form of the protein exists (soluble CD83 [sCD83]) (8). The structural analysis revealed that CD83 is well conserved soluble protein was detected in the blood of healthy donors and T by guest on September 28, 2021 among mammalian species. The murine cd83 gene consists of five elevated sCD83 levels in patients suffering from hematological exons, and the protein shares 63% sequence homology to the malignancies and rheumatoid arthritis (9, 10). Moreover, sCD83 human protein (2). CD83 has been described as one of the most was described to be released from activated DCs, B cells (8), and prominent marker molecules to identify mature dendritic cells stimulated CD83+ T cells (11). Several studies provide evidence (DCs) (3, 4). However, other immune cells including activated for sCD83 having immunosuppressive functions. Treatment of B cells and T cells have been shown to express CD83 (5–7). DCs or monocytes with sCD83 resulted in impaired maturation and T cell stimulatory function in vitro (12–15). Strikingly, ap- plication of sCD83 to mice ameliorates experimental colitis (16), *Institute of Medical Microbiology, University Hospital Essen, University Duisburg- Essen, 45147 Essen, Germany; †Institute for Medical Medicine, University Medical prevents the development of experimental autoimmune enceph- Center of the Johannes-Gutenberg University Mainz, 55131 Mainz, Germany; alitis (EAE) (13), alleviates experimental autoimmune uveitis ‡Institute of Veterinary Pathology, Free University of Berlin, 14163 Berlin, Germany; x (17), and has protective effects in different transplantation settings Transgenic Core Facility, Max Planck Institute of Molecular Cell Biology and Genetics, 01307 Dresden, Germany; {Max Planck Institute for Metabolism Research, (18, 19). 2 2 Center for Endocrinology, Diabetes and Preventive Medicine, University Hospital of / ‖ Analysis of CD83 mice and B cell–specific CD83-deficient Cologne, University of Cologne, 50931 Cologne, Germany; and Excellence Cluster mice revealed that CD83 is involved in B cell activation. Loss of on Cellular Stress Responses in Aging Associated Diseases, University of Cologne, 50931 Cologne, Germany CD83 expression in B cells resulted in downregulation of MHC ORCIDs: 0000-0002-6308-0568 (R.K.); 0000-0003-2084-5150 (R.N.). class II (MHC-II) and CD86 as well as decreased proliferation Received for publication January 13, 2020. Accepted for publication April 10, 2020. upon stimulation (20, 21). Stabilization of MHC-II and CD86 by CD83 was also reported for thymus cortical epithelial cells (22) This work was supported by the Deutsche Forschungsgemeinschaft (HA5774/3) to W.H. and DCs (23). These results suggest that CD83 is involved in + Address correspondence and reprint requests to Prof. Wiebke Hansen, Institute of CD4 T cell development and priming by modulating the function 2 2 Medical Microbiology, University Hospital Essen, University Duisburg-Essen, of APCs in the thymus and periphery. Indeed, CD83 / mice Hufelandstraße 55, 45147 Essen, Germany. E-mail address: wiebke.hansen@uk- showed tremendously reduced frequencies of peripheral CD4+ essen.de T cells (24), and downregulation of CD83 on human DCs resulted The online version of this article contains supplemental material. in impaired T cell stimulation in an allogenic MLR (25). Abbreviations used in this article: CD83cKO, CD83flox/flox/CD4-cretg/wt; CD83cWT, CD83flox/flox/CD4-crewt/wt; CHS, contact hypersensitivity; DC, dendritic cell; DNFB, It is well established that regulatory T cells (Tregs) play an 2,4-dinitrofluorobenzene; EAE, experimental autoimmune encephalitis; MHC-II, important role in the maintenance of immune homeostasis and MHC class II; mLN, mesenteric lymph node; sCD83, soluble CD83; Treg, regulatory control of immune responses against pathogens and tumor cells T cell; WT, wild-type. because of their immunosuppressive function (26–28). We iden- + Copyright Ó 2020 by The American Association of Immunologists, Inc. 0022-1767/20/$37.50 tified CD83 to be highly expressed by Foxp3 Tregs in comparison www.jimmunol.org/cgi/doi/10.4049/jimmunol.2000042 2 ENDOGENOUS CD83 EXPRESSION REGULATES T CELL RESPONSES 2 with Foxp3 T cells (5). In accordance, CD83+ T cells from Scientific) following the manufacturer’s instructions and stimulated with 1 mg/ml anti-CD3 in the presence of 3 3 105 irradiated splenocytes from CD83-eGFP reporter mice have been shown to express Treg- + associated molecules and to have inhibitory activity in vitro as Thy1.1 BALB/c mice as APCs for 72 h. Proliferation was assessed as loss of eFluo670 on gated CD4+Thy1.2+ T cells by flow cytometry. To deter- well as during experimental colitis in vivo (11). Overexpression of mine the suppressive capacity CD4+CD25+ Tregs were isolated by cell murine CD83 by retroviral gene transfer conferred a suppressive sorting and cocultured with Cell Proliferation Dye eFluor 670–labeled 2 phenotype to CD4+CD252 T cells in vitro and adoptive transfer of CD4+CD25 responder T cells from Thy1.1 mice in the presence of these CD83-overexpressing T cells ameliorated the development 1 mg/ml anti-CD3 and irradiated splenocytes for 3 d. Proliferation was assessed as loss of eFluor 670 on gated CD4+Thy1.1+ responder T cells by of EAE and contact hypersensitivity (CHS) reaction in vivo (5). flow cytometry. Most recently, similar results were described for T cells from bacterial artificial chromosome transgenic mice expressing human Differentiation of Th1 and induced Tregs + 2 CD83. CD4 T cells from these mice exhibited reduced prolifer- A total of 1 3 106 sorted CD4+CD25 T cells were stimulated with ative activity, and transgenic mice recover faster from EAE than 5 mg/ml anti-CD3 (BD Biosciences) plate-bound and 1 mg/ml anti-CD28 wild-type (WT) mice (29). These studies indicate that enforced (BD Biosciences) (Th0) either in the presence of 100 U/ml IL-2 (Thermo Fisher Scientific) and 5 ng/ml rTGF-b (R&D Systems) for CD83 expression modulates the function of CD4+ T cells. + 3 d (induced Treg), in the presence of 20 ng/ml rIL-12 (R&D Systems) and However, the impact of endogenous CD83 on the CD4 Tcell 200 ng/ml anti–IL-4 (Thermo Fisher Scientific) for 5 d (Th1) or in the 2 phenotype, in particular on CD4+Foxp3 conventional T cells, presence of 2 ng/ml rTGF-b, 50 ng/ml rIL-6, 20 ng/ml rIL-1b, 100 ng/ml still remains elusive.