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Keratinocyte-Intrinsic MHCII Expression Controls Microbiota-Induced Th1 Cell Responses

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Keratinocyte-Intrinsic MHCII Expression Controls Microbiota-Induced Th1 Cell Responses Keratinocyte-intrinsic MHCII expression controls microbiota-induced Th1 cell responses Samira Tamoutounoura,1,2, Seong-Ji Hana,1, Julie Deckersb,c, Michael G. Constantinidesa, Charlotte Hurabiellea,d,3, Oliver J. Harrisona, Nicolas Bouladouxa,e, Jonathan L. Linehana,4, Verena M. Linka, Ivan Vujkovic-Cvijina, Paula Juliana Perez-Chaparroe, Stephan P. Rosshartf,5, Barbara Rehermannf, Vanja Lazarevicg, and Yasmine Belkaida,e,6 aMetaorganism Immunity Section, Laboratory of Immune System Biology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892; bImmunoregulation Unit, Vlaams Instituut voor Biotechnologie Center for Inflammation Research, 9052 Ghent, Belgium; cDepartment of Internal Medicine, Ghent University, 9000 Ghent, Belgium; dInserm U976, Hôpital Saint Louis, Université Paris Diderot, 75010 Paris, France; eNational Institute of Allergy and Infectious Diseases Microbiome Program, National Institutes of Health, Bethesda, MD 20892; fImmunology Section, Diseases Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892; and gExperimental Immunology Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892 Contributed by Yasmine Belkaid, September 21, 2019 (sent for review July 22, 2019; reviewed by Florent Ginhoux and Brian Kelsall) The cross-talk between the microbiota and the immune system to which epithelial cells from distinct compartments respond to plays a fundamental role in the control of host physiology. commensals in a way that supports defined classes of adaptive However, the tissue-specific factors controlling this dialogue remain immunity remains poorly understood. poorly understood. Here we demonstrate that T cell responses to Here we found that the ability of commensal-specific Th1 cells commensal colonization are associated with the development of to accumulate within the skin was associated with the physical organized cellular clusters within the skin epithelium. These orga- organization of T cells within epidermal lymphocyte clusters. nized lymphocyte clusters are surrounded by keratinocytes express- Within these clusters, interferon-γ (IFN-γ)productionby ing a discrete program associated with antigen presentation and + antimicrobial defense. Notably, IL-22–mediated keratinocyte-intrinsic CD4 T cells was controlled by major histocompatibility complex MHC class II expression was required for the selective accumulation class II (MHCII)-expressing keratinocytes. Taken together, our of commensal-induced IFN-γ, but not IL-17A–producing CD4+ Tcells work uncovers a tissue checkpoint involved in the discrete control INFLAMMATION within the skin. Taking these data together, this work uncovers an IMMUNOLOGY AND unexpected role for MHC class II expression by keratinocytes in the Significance control of homeostatic type 1 responses to the microbiota. Our find- ings have important implications for the understanding of the tissue- The cross-talk between the microbiota and the immune system specific rules governing the dialogue between a host and its plays a fundamental role in the control of host physiology. microbiota. However, the tissue-specific factors controlling this dialogue remain poorly understood. Here we demonstrate that T cell microbiota | skin | keratinocyte | antigen presentation | Th1 responses to commensal colonization are associated with the development of organized cellular clusters within the skin ep- he microbiota play a fundamental role in the induction and ithelium. Keratinocyte-intrinsic MHC class II expression was Tquality of local immune responses (1). A large fraction of this required for the selective accumulation of commensal-induced + response is directed at the microbiota itself (1, 2). Notably, IFN-γ–producing CD4 T cells within the skin. This work un- adaptive recognition of the microbiota controls several aspects of covers an unexpected role for MHC class II expression by tissue physiology, including antimicrobial defense and tissue re- keratinocytes in the control of homeostatic type 1 responses to pair (1–4). As such, identification of the factors controlling the the microbiota. Our findings have important implications for quality and function of commensal-induced T cells has important the understanding of the tissue-specific rules governing the implications for our understanding of host immunity and tissue dialogue between a host and its microbiota. pathologies. Commensal-specific T cells exert their local function via the Author contributions: S.T. and Y.B. designed research; S.T., S.-J.H., J.D., M.G.C., C.H., release of defined cytokines, such as interleukin (IL)-17A and O.J.H., N.B., J.L.L., V.M.L., I.V.-C., and P.J.P.-C. performed research; S.P.R., B.R., and V.L. contributed new reagents/analytic tools; S.T., S.-J.H., J.D., M.G.C., C.H., O.J.H., N.B., J.L.L., IL-22, the production of which is tightly regulated by local cues V.M.L., I.V.-C., and Y.B. analyzed data; and S.T., S.-J.H., and Y.B. wrote the paper. (1). Indeed, adaptive immunity to commensal microbes relies Reviewers: F.G., Singapore Immunology Network; and B.K., National Institute of Allergy on a highly controlled sequence of events, including functional and Infectious Diseases. licensing by tissue-specific factors or mechanisms. Epithelial The authors declare no competing interest. cells, as the first-line in the host microbiota dialogue, play a Published under the PNAS license. critical role in these processes. For example, within the gastro- Data deposition: The data reported in this paper have been deposited in the Gene Ex- intestinal tract, segmented filamentous bacteria promote the pression Omnibus (GEO) database, https://www.ncbi.nlm.nih.gov/geo (accession no. production of serum amyloid A (SAA) by intestinal epithelial GSE125657). cells. SAA in turn licenses IL-17A production by segmented 1S.T. and S.-J.H. contributed equally to this work. + filamentous bacteria-specific CD4 T cells (5). Furthermore, in 2Present address: L’Oréal Research and Innovation, 93600 Aulnay-sous-Bois, France. — the gut, epithelial cell responses to the microbiota particularly 3Present address: Internal Medicine Department, University of California, San Francisco, the production of IL-18, IL-25, IL-33, and thymic stromal lym- CA 94143. — phopoietin have been shown to control both the frequency and 4Present address: Department of Cancer Immunology, Genentech, South San Francisco, function of innate lymphoid cells (ILCs) (6). Evidence also CA 94080. supports the idea that commensal-induced cues are tissue- 5Present address: Klinik für Innere Medizin II, University of Freiburg, 79106 Freiburg, specific. For example, within the skin, the ability of the micro- Germany. biota to promote local production of IL-1α and IL-23 licenses 6To whom correspondence may be addressed. Email: [email protected]. cutaneous T cells to release IL-17A (7, 8). However, much re- This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10. mains to be learned about how defined microenvironments can 1073/pnas.1912432116/-/DCSupplemental. shape the host–microbiota dialogue. More particularly, the extent www.pnas.org/cgi/doi/10.1073/pnas.1912432116 PNAS Latest Articles | 1of10 Downloaded by guest on September 25, 2021 ACIL-17AYFP CD8 CD49f CD8 CD4 MHCII+CD49f+ CD8 CD4 HF HF HF HF HF HF 30 m HF CD4 IL-17AYFP CD8 CD4 CD49f 100 m HF CD8 CD4 MHCII+CD49f+ MHCII+CD49f+ 100 m 100 m HF HF B T-betZsGreen CD8 CD8 CD4 MHCII+CD49f+ CD8 CD4 HF HF 20 m CD4 T-betZsGreen CD8 CD4 CD49f 50 m 50 m HF HF E F 10 * D Control S. epidermidis 8 105 105 inocytes (%) 6 t 104 104 4 MHCII+ keratinocytes 103 103 kera CD49f 0.4±0.2 3.3±1.3 + 2 95.3±2.2 Interfollicular U 102 102 1.9±1.1 Infundibulum 0 0 Ed 0 MHCII keratinocytes 0.5±0.1 Isthmus 0103 104 105 0103 104 105 MHCII keratinocytes MHCII ) % G ( 5 *** es *** Interfollicular 4 cyt Infundibulum ino 3 Isthmus Supra- 2 basal kerat + Basal bulge bulge 1 0 MHCII HIJ KL * Germ-free ) 6 ** 8 10 3 %) ** (%) (% 6 ** * *** ** ( * es 6 8 es 4 2 ytes 6 c 4 inocyt 4 inocyt t 4 tino 2 1 2 erat 2 era keratinocytes (%) k kera k keratinocytes (%) 2 + + + + + 0 0 0 0 0 MHCII MHCII Ctrl MHCII Ctrl Ctrl MHCII MHCII Ctrl S. epi C. alb Wild LM061LM087 S. aur B6 SPFB6 Conv NIH05001 Fig. 1. S. epidermidis induces MHCII+ keratinocytes and lymphocyte clusters in the epidermis. (A) Representative confocal microscopy image of a projection on the z axis of an Il17aCreRosa26YFP (IL-17AYFP) mouse epidermis stained for CD8α, CD4, and CD49f at day 14 postassociation with S. epidermidis. HF: Hair follicle. (Scale bars, 30 μm.) (B) Representative confocal microscopy image of a projection on the z axis of a T-betZsGreen mouse epidermis stained for CD8α, CD4, and CD49f at day 14 postassociation with S. epidermidis. HF: Hair follicle. (Scale bars, 20 μm.) (C) Representative confocal microscopy image of C57BL/6 + + mouse epidermis stained for CD8α, CD4, MHCII, and CD49f at day 14 postassociation with S. epidermidis. MHCII CD49f keratinocytes result from colocali- + + + + zation between MHCII and CD49f cells. (Scale bars, 100 μmor50μm.) Dotted orange lines show the delineation of the areas containing CD49f MHCII + keratinocytes. (D) Flow cytometric analysis of MHCII expression by epidermal CD49f keratinocytes from unassociated control mice or mice at day 14 post- association with S. epidermidis.(E) Frequencies of EdU incorporation by MHCII+ keratinocytes and MHCII− keratinocytes at 14 d after S. epidermidis asso- ciation. (F) Frequencies of MHCII+ keratinocyte populations by indicated cellular phenotype. (G) Schematic illustrating the anatomical localization of distinct + keratinocyte populations. Graph represents the frequencies of MHCII expression by distinct keratinocyte populations. (H–K) Frequencies of epidermal MHCII keratinocytes in control (Ctrl) unassociated mice or in mice associated with different strains (LM061, LM087, NIH05001) of S. epidermidis (H), in S. epidermidis + (LM087)-associated germ-free mice (I), and in mice associated with S. aureus (J)orC.
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