Plasticity of Th17 Cells in Autoimmune Kidney Diseases Christian F. Krebs, Jan-Eric Turner, Hans-Joachim Paust, Sonja Kapffer, Tobias Koyro, Sonja Krohn, Friederike Ufer, This information is current as Manuel A. Friese, Richard A. Flavell, Brigitta Stockinger, of October 2, 2021. Oliver M. Steinmetz, Rolf A. K. Stahl, Samuel Huber and Ulf Panzer J Immunol 2016; 197:449-457; Prepublished online 6 June 2016; doi: 10.4049/jimmunol.1501831 Downloaded from http://www.jimmunol.org/content/197/2/449 Supplementary http://www.jimmunol.org/content/suppl/2016/06/04/jimmunol.150183 http://www.jimmunol.org/ Material 1.DCSupplemental References This article cites 35 articles, 14 of which you can access for free at: http://www.jimmunol.org/content/197/2/449.full#ref-list-1 Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision by guest on October 2, 2021 • 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 © 2016 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology Plasticity of Th17 Cells in Autoimmune Kidney Diseases Christian F. Krebs,* Jan-Eric Turner,* Hans-Joachim Paust,* Sonja Kapffer,* Tobias Koyro,* Sonja Krohn,* Friederike Ufer,† Manuel A. Friese,† Richard A. Flavell,‡ Brigitta Stockinger,x Oliver M. Steinmetz,* Rolf A. K. Stahl,* Samuel Huber,{ and Ulf Panzer* The ability of CD4+ T cells to differentiate into pathogenic Th1 and Th17 or protective T regulatory cells plays a pivotal role in the pathogenesis of autoimmune diseases. Recent data suggest that CD4+ T cell subsets display a considerable plasticity. This plasticity seems to be a critical factor for their pathogenicity, but also for the potential transition of pathogenic effector T cells toward a more tolerogenic phenotype. The aim of the current study was to analyze the plasticity of Th17 cells in a mouse model of acute crescentic glomerulonephritis and in a mouse chronic model of lupus nephritis. By transferring in vitro generated, highly purified Th17 cells and by using IL-17A fate reporter mice, we demonstrate that Th17 cells fail to acquire substantial expression of the Th1 and Th2 signature cytokines IFN-g and IL-13, respectively, or the T regulatory transcription factor Foxp3 throughout the course Downloaded from of renal inflammation. In an attempt to therapeutically break the stability of the Th17 phenotype in acute glomerulonephritis, we subjected nephritic mice to CD3-specific Ab treatment. Indeed, this treatment induced an immunoregulatory phenotype in Th17 cells, which was marked by high expression of IL-10 and attenuated renal tissue damage in acute glomerulonephritis. In summary, we show that Th17 cells display a minimum of plasticity in acute and chronic experimental glomerulonephritis and introduce anti- CD3 treatment as a tool to induce a regulatory phenotype in Th17 cells in the kidney that may be therapeutically exploited. The Journal of Immunology, 2016, 197: 449–457. http://www.jimmunol.org/ he CD4+ T cells play a central role in the pathogenesis glomerulonephritis (i.e., nephrotoxic nephritis [NTN]) (8–10) and of autoimmune diseases, including human and experi- a model of pristane-induced lupus nephritis (11, 12). Time course T mental crescentic glomerulonephritis. Classically, it was analyses of the renal CD4+ T cell response in the NTN model assumed that the cell-mediated immune reactions responsible for revealed that Th17 cell numbers peak at ∼days 6–10 after ne- tissue injury are predominantly mediated by IFN-g–producing phritis induction and then decline, whereas the numbers of Th1 Th1 cells (1, 2). However, the identification and functional char- and T regulatory (Treg) cells continuously increase from days 6 to acterization of Th17 cells have challenged this paradigm (3, 4). 10 onward (13–16). This sequential infiltration of the different by guest on October 2, 2021 Indeed, numerous studies have clearly shown that Th17 cells drive CD4+ T cell subsets into inflamed target organs with an early autoimmune processes that were previously thought to be exclu- increase of Th17 cells that is followed by Th1 and Treg cell in- sively Th1 mediated, such as multiple sclerosis (5), rheumatoid filtration is a phenomenon that has also been reported in other arthritis (6), and psoriasis (7). inflammatory diseases (17, 18). Whether the gradual decline of We and others recently demonstrated that the IL-17A/Th17 axis Th17 cells observed during progression of renal autoimmunity is contributes to renal tissue injury in both experimental crescentic due to transdifferentiation into cells of another Th cell subtype, a result of apoptosis, or a consequence of emigration of Th17 cells is currently unclear. + *III. Medizinische Klinik, Universita¨tsklinikum Hamburg-Eppendorf, 20246 Ham- In this context, recent studies indicating that CD4 Tcell † burg, Germany; Institute of Neuroimmunology and Multiple Sclerosis, Universita¨ts- subsets in other organs demonstrate a higher grade of plastic- klinikum Hamburg-Eppendorf, 20246 Hamburg, Germany; ‡Department of Immunobiology, School of Medicine, Yale University, New Haven, CT 06520; ity than previously appreciated are of great interest (18–21). In xDivision of Molecular Immunology, Medical Research Council National Institute { particular, Th17 and Treg cells might be highly flexible, which for Medical Research, London NW7 1AA, United Kingdom; and I. Medizinische would allow these cells to switch to a different Th cell program Klinik, Universita¨tsklinikum Hamburg-Eppendorf, 20246 Hamburg, Germany and thereby adapt to various inflammatory settings during an ORCIDs: 0000-0002-8412-1840 (T.K.); 0000-0001-8781-336X (B.S.); 0000-0002- 6225-8600 (R.A.K.S.). immune response. However, whether the potential plasticity of Received for publication August 24, 2015. Accepted for publication May 4, 2016. Th17 cells in autoimmunity is a general finding, and whether this process is beneficial or detrimental remains to be fully This work was supported by Deutsche Forschungsgemeinschaft Grants SFB1192/A5 (to C.F.K. and S.H.), SFB1192/A1 (to U.P.), and KFO 228: PA 754/7-2 (to U.P. and elucidated. C.F.K.); Emmy-Noether-Programme Grants TU316/2-1 (to J.-E.T.) and HU1714/3-1 The aim of the current study was to investigate the potential (to S.H.); and grants from the Deutsche Gesellschaft fur€ Nephrologie (to C.F.K. and J.-E.T.). T.K. received a stipend from the Werner Otto Foundation. plasticity of Th17 cells in murine models of glomerulonephritis. We therefore did the following: 1) assessed the fate of adoptively Address correspondence and reprint requests to Dr. Christian F. Krebs, Universita¨ts- klinikum Hamburg-Eppendorf, III. Medizinische Klinik, Martinistraße 52, 20246 transferred Th17 cells in nephritic Rag1-knockout mice; 2) studied Hamburg, Germany. E-mail address: [email protected] the plasticity of in vivo differentiated Th17 cells in two models of The online version of this article contains supplemental material. glomerulonephritis (NTN and pristane-induced lupus nephritis) by Cre YFP Abbreviations used in this article: EAE, experimental autoimmune encephalomyeli- using IL-17A fate reporter mice (Il17a 3 R26r ); and 3) tis; GN, glomerulonephritis; NTN, nephrotoxic nephritis; PAS, periodic acid–Schiff; modulated Th17 cell plasticity by CD3-specific Ab treatment to Tr1, type 1 regulatory; Treg, T regulatory. induce a regulatory phenotype of Th17 cells in crescentic glo- Copyright Ó 2016 by The American Association of Immunologists, Inc. 0022-1767/16/$30.00 merulonephritis (GN). www.jimmunol.org/cgi/doi/10.4049/jimmunol.1501831 450 Th17 CELL PLASTICITY IN GN Materials and Methods Ag retrieval was performed by incubation with proteinase type XXIV (5 mg/ml; Animals Sigma-Aldrich) for 15 min at 37˚C. Tissue sections were developed with the Vectastain ABC-AP kit (Vector Laboratories, Burlingame, CA). To All mice in this study were on C57BL/6J background. Il-17aGFP/Ifn-gFP635/ quantify tubulointerstitial GR1+ cells, at least 20 low power fields (original Foxp3mRFP and Il-10GFP reporter mice as well as IL-17ACre 3 R26rYFP magnification 3200) were counted. mice were used, as described before (18, 22–24). Rag12/2 mice were All slides were evaluated using an Axioskop light microscope (Zeiss) and purchased from The Jackson Laboratory (Bar Harbor, ME). All mice were photographed with an Axiocam HRc (Zeiss) using the ZEN lite 2011 raised under specific pathogen-free conditions in our animal facility at the software (Zeiss). University Medical Center Hamburg-Eppendorf. Animal experiments were performed according to national animal care and ethical guidelines, and Isolation of leukocytes from kidney, spleen, and CNS were approved by local ethics committees. Previously described methods for leukocyte isolation from murine kidneys Animal models and functional studies were used (9). In brief, kidneys were digested for 45 min at 37˚C with 0.4 mg/ml collagenase D (Roche, Mannheim, Germany) and 0.01 mg/ml NTN was induced in 8- to 10-wk-old male mice by i.p. injection of 0.5 ml DNase I in RPMI 1640 medium (Life Technologies, Darmstadt, Ger- nephrotoxic sheep serum per mouse, as previously described (10). In many) supplemented with 10% heat-inactivated FCS (Invitrogen, Carlsbad, adoptive cell transfer experiments, cells were administered i.v. 1 d prior to CA), and finely minced using GentleMACS digester (Miltenyi Biotec, induction of glomerulonephritis i.v. Lupus nephritis was induced by i.p. Bergisch-Gladbach, Germany).