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IL-7Rα Expression Regulates Murine Dendritic Cell Sensitivity to Thymic Stromal Lymphopoietin

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IL-7Rα Expression Regulates Murine Dendritic Cell Sensitivity to Thymic Stromal Lymphopoietin IL-7Rα Expression Regulates Murine Dendritic Cell Sensitivity to Thymic Stromal Lymphopoietin This information is current as Laura Kummola, Zsuzsanna Ortutay, Xi Chen, Stephane of September 28, 2021. Caucheteux, Sanna Hämäläinen, Saara Aittomäki, Ryoji Yagi, Jinfang Zhu, Marko Pesu, William E. Paul and Ilkka S. Junttila J Immunol 2017; 198:3909-3918; Prepublished online 12 April 2017; Downloaded from doi: 10.4049/jimmunol.1600753 http://www.jimmunol.org/content/198/10/3909 Supplementary http://www.jimmunol.org/content/suppl/2017/04/12/jimmunol.160075 http://www.jimmunol.org/ Material 3.DCSupplemental References This article cites 32 articles, 17 of which you can access for free at: http://www.jimmunol.org/content/198/10/3909.full#ref-list-1 Why The JI? Submit online. by guest on September 28, 2021 • 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 © 2017 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology IL-7Ra Expression Regulates Murine Dendritic Cell Sensitivity to Thymic Stromal Lymphopoietin Laura Kummola,*,1 Zsuzsanna Ortutay,*,1 Xi Chen,† Stephane Caucheteux,†,2 Sanna Ha¨ma¨la¨inen,‡ Saara Aittoma¨ki,‡ Ryoji Yagi,†,3 Jinfang Zhu,† Marko Pesu,‡,x William E. Paul,†,1,4 and Ilkka S. Junttila*,{,1 Thymic stromal lymphopoietin (TSLP) and IL-7 are related cytokines that mediate growth and differentiation events in the immune system. They signal through IL-7Ra–containing receptors. Target cells of TSLP in Th2 responses include CD4 T cells and dendritic cells (DCs). Although it has been reported that expression of TSLP receptor (TSLPR) on CD4 T cells is required for OVA-induced lung inflammation, DCs have also been shown to be target cells of TSLP. In this study, we show that murine ex vivo splenic DCs are unresponsive to TSLP, as they fail to phosphorylate STAT5, but in vitro overnight culture, especially in presence of IL-4, renders DCs responsive to both TSLP and IL-7. This induced responsiveness is accompanied by dramatic upregulation of Downloaded from 2 IL-7Ra on DCs with little change in expression of TSLPR or of gc. In splenic DCs, the induction of IL-7Ra occurs mainly in CD8 DCs. In vivo, we found that IL-4 has a differential regulatory role on expression of IL-7Ra depending on the cell type; IL-4 decreases IL-7Ra expression on CD4 T cells whereas it upregulates the expression on DCs. Our results indicate that the induction of IL-7Ra expression on DCs is critical for TSLP responsiveness and that IL-4 can upregulate IL-7Ra on DCs. The Journal of Immunology, 2017, 198: 3909–3918. http://www.jimmunol.org/ ignaling by thymic stromal lymphopoietin (TSLP) and IL-7 as well as DNA binding of STAT5, resulting in regulation of depends on ligand-mediated assemblage of a receptor STAT5-mediated transcription. In contrast, TSLP binds TSLP S complex consisting of a cytokine-specific binding chain receptor (TSLPR), a close homolog of gc,followedbyIL-7Ra and an auxiliary chain. The TSLP and IL-7 receptors share IL-7Ra, recruitment to the complex. The KD for TSLP binding to TSLPR 9 although they use this chain distinctly. IL-7 binds with high af- is ∼7 3 10 (2), whereas the KD for IL-7 binding to IL-7Ra is 1 3 finity to IL-7Ra, and this complex then binds common g chain 1010 (3), indicating that initial cytokine binding to ligand-binding (gc), activating the Jak1/Jak3 kinases, followed by phosphoryla- receptor chain occurs with higher affinity by IL-7 than by TSLP. tion of critical tyrosine residues on IL-7Ra (1). Docking of STAT5 As opposed to IL-7, the Jak1 and Jak2 kinases are activated by by guest on September 28, 2021 to the phosphorylated tyrosine residues of IL-7Ra results in TSLP in human and mouse primary CD4 T cells with the con- Jak-mediated phosphorylation and subsequent nuclear translocation, sequent phosphorylation of STAT5 (1). In human myeloid den- dritic cells (DCs), activation of STAT6 by TSLP has also been *Faculty of Medicine and Life Sciences, University of Tampere, 33520 Tampere, Fin- reported (4). land; †Laboratory of Immunology, National Institute of Allergy and Infectious Diseases, Functionally, IL-7 is a critical growth factor for B and T cell National Institutes of Health, Bethesda, MD 20892; ‡Biomeditech, University of Tam- x development and induces survival of memory T cells (5). Re- pere, 33014 Tampere, Finland; Department of Dermatology, Tampere University Hos- pital, 33521 Tampere, Finland; and {Fimlab Laboratories, 33520 Tampere, Finland cently, the role of IL-7 in development of innate lymphoid cells 1L.K., Z.O., W.E.P., and I.S.J. contributed equally to this work. has also been established (6). It is of interest that epithelial cells 2Current address: Institute of Infection and Immunity, Cardiff University, Cardiff, U.K. possess the capacity to produce IL-7, which might be important 3 for survival of tissue lymphocytes locally (7). Memory cell Current address: Department of Immunology, Graduate School of Medicine, Chiba University, Chiba, Japan. competition for IL-7 has been suggested to be the mechanism of 4Deceased. regulation of IL-7–mediated growth (8). ORCIDs: 0000-0001-7621-2753 (R.Y.); 0000-0002-7541-3921 (M.P.). Although mast cells (9), basophils (10), and DCs (11) have been Received for publication April 27, 2016. Accepted for publication March 13, 2017. reported to produce TSLP, the main TSLP-producing cells appear to be the epithelial cells in humans and particularly the kerati- This work was supported by the National Institute of Allergy and Infectious Diseases Intramural Research Program (to X.C., S.C., R.Y., W.E.P., J.Z., and I.S.J.), the Finn- nocytes in mice (12–14). Thus, TSLP appears to be principally ish Medical Foundation (to I.S.J.), the Sigrid Juselius Foundation (to I.S.J. and M.P.), expressed at skin and mucosal barriers; in contrast, IL-7 produc- the Tampere Tuberculosis Foundation (to I.S.J. and M.P.), Competitive State Re- search Financing of the Expert Responsibility Area of Tampere University Hospital tion seems more a property of stromal cells in primary and Grants 9M080, 9N056, and 9S051 (to M.P.), Fimlab Laboratories Grant X51409 (to secondary lymphoid organs (15). Effectively, this suggests a I.S.J.), Academy of Finland Projects 263955 and 135980 (to M.P.), and by the Emil specialization of the functions of the two cytokines and particu- Aaltonen Foundation (to L.K. and M.P.). larly emphasizes the potential importance of TSLP in action at Address correspondence and reprint requests to Dr. Ilkka S. Junttila, Faculty of Medicine and Life Sciences, University of Tampere, Room F354 Arvo-Building, mucosal surfaces, be it in the induction of Ag-presenting com- La¨a¨ka¨rinkatu 1, FI-33520 Tampere, Finland. E-mail address: Ilkka.junttila@uta.fi petence in DCs or in the differentiation/expansion of memory/ The online version of this article contains supplemental material. effector T cells in tissues. Abbreviations used in this article: gc, common g chain; DC, dendritic cell; MHC II, To respond to TSLP, target cells need to express both IL-7Ra and MHC class II; o/n, overnight; TSLP, thymic stromal lymphopoietin; TSLPR, TSLP TSLPR, whereas those responsive to IL-7 require expression of receptor. IL-7Ra and gc (16). Of conventional or plasmacytoid DCs Copyright Ó 2017 by The American Association of Immunologists, Inc. 0022-1767/17/$30.00 from unmanipulated mice, only migratory and skin DCs express www.jimmunol.org/cgi/doi/10.4049/jimmunol.1600753 3910 IL-7Ra EXPRESSION ON DCs IL-7Ra at a low level, although IL-7Ra expression appears to be under ethical permission R21002 from the Ethics Committee of Pirkan- important for DC development (17). Human myeloid DCs have maa Hospital District using pan-human DC purification kit (Miltenyi been reported to alter their behavior in response to TSLP by di- Biotec). All reagents used were provided endotoxin free by the manu- facturers and where applicable they were dissolved into sterile PBS rectly upregulating CD40 and CD80 and by enhancing their ca- containing BSA (low endotoxin; Sigma-Aldrich, St. Louis, MO). Strict pacity to induce T cell proliferation and chemotaxis, with the aseptic techniques in biosafety level 2 sterile tissue culture hoods were latter through expression of CCL17 and CCL22 (12, 18). Freshly used in all experiments. isolated human myeloid DCs express low levels of both IL-7Ra and TSLPR, and overnight (o/n) culture of these cells strongly Abs, cytokines, flow cytometry, and statistics upregulates expression of both receptor chains (19). Recently, Mouse Abs and isotype controls were either from BD Biosciences (Franklin basophils were shown to be responsive to TSLP, and it has been Lakes, NJ; pSTAT5, B220), eBioscience (Santa Clara, CA; CD11c, CD49/ reported that such basophils acquire a Th2 phenotype in that they Dx5, MHC class II [MHC II], CD3, CD4, CD8, F4/80, IL-7Ra, gc, CD80, produce more IL-4 in response to cytokine stimulation than do CD86, isotype controls for IL-7Ra and gc), or R&D Systems (Minneap- olis, MN; TSLPR and polyclonal goat Ig as a control).
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