Abstract

The mechanisms by which mucosal homeostasis is maintained are of central importance to inflammatory bowel disease. Critical to these processes is the intestinal epithelial cell (IEC), which regulates immune responses at the interface between the commensal microbiota and the host1,2. CD1d presents self and microbial lipid antigens to natural killer T (NKT) cells, which are involved in the pathogenesis of colitis in animal models and human inflammatory bowel disease3,4,5,6,7,8. As CD1d crosslinking on model IECs results in the production of the important regulatory cytokine interleukin (IL)-10 (ref. 9), decreased epithelial CD1d expression—as observed in inflammatory bowel disease10,11—may contribute substantially to intestinal inflammation. Here we show in mice that whereas bone-marrow-derived CD1d signals contribute to NKT-cell-mediated intestinal inflammation, engagement of epithelial CD1d elicits protective effects through the activation of STAT3 and STAT3-dependent transcription of IL-10, heat shock protein 110 (HSP110; also known as HSP105), and CD1d itself. All of these epithelial elements are critically involved in controlling CD1d-mediated intestinal inflammation. This is demonstrated by severe NKT-cell-mediated colitis upon IEC-specific deletion of IL-10, CD1d, and its critical regulator microsomal triglyceride transfer protein (MTP)12,13, as well as deletion of HSP110 in the radioresistant compartment. Our studies thus uncover a novel pathway of IEC-dependent regulation of mucosal homeostasis and highlight a critical role of IL-10 in the intestinal epithelium, with broad implications for diseases such as inflammatory bowel disease.

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References

  1. 1.

    , & Inflammatory bowel disease. Annu. Rev. Immunol. 28, 573–621 (2010)

  2. 2.

    , & Welcome to the neighborhood: epithelial cell-derived cytokines license innate and adaptive immune responses at mucosal sites. Immunol. Rev. 226, 172–190 (2008)

  3. 3.

    et al. Nonclassical CD1d-restricted NK T cells that produce IL-13 characterize an atypical Th2 response in ulcerative colitis. J. Clin. Invest. 113, 1490–1497 (2004)

  4. 4.

    , , , & Oxazolone colitis, a Th2 colitis model resembling ulcerative colitis, is mediated by IL-13-producing NK-T cells. Immunity 17, 629–638 (2002)

  5. 5.

    et al. Microbial exposure during early life has persistent effects on natural killer T cell function. Science 336, 489–493 (2012)

  6. 6.

    & Role of NKT cells in the digestive system. IV. The role of canonical natural killer T cells in mucosal immunity and inflammation. Am. J. Physiol. Gastrointest. Liver Physiol. 294, G1–G8 (2008)

  7. 7.

    et al. Interleukin-13 is the key effector Th2 cytokine in ulcerative colitis that affects epithelial tight junctions, apoptosis, and cell restitution. Gastroenterology 129, 550–564 (2005)

  8. 8.

    et al. dysregulation of CD1d-restricted type II natural killer T cells leads to spontaneous development of colitis in mice. Gastroenterology 142, 326–334 (2012)

  9. 9.

    , , & Ligation of intestinal epithelial CD1d induces bioactive IL-10: critical role of the cytoplasmic tail in autocrine signaling. Proc. Natl Acad. Sci. USA 96, 13938–13943 (1999)

  10. 10.

    et al. Expression of nonclassical class I molecules by intestinal epithelial cells. Inflamm. Bowel Dis. 13, 298–307 (2007)

  11. 11.

    et al. Expression of a nonpolymorphic MHC class I-like molecule, CD1D, by human intestinal epithelial cells. J. Immunol. 147, 2518–2524 (1991)

  12. 12.

    et al. CD1d function is regulated by microsomal triglyceride transfer protein. Nature Med. 10, 535–539 (2004)

  13. 13.

    et al. Primary deficiency of microsomal triglyceride transfer protein in human abetalipoproteinemia is associated with loss of CD1 function. J. Clin. Invest. 120, 2889–2899 (2010)

  14. 14.

    et al. Tissue-specific and inducible Cre-mediated recombination in the gut epithelium. Genesis 39, 186–193 (2004)

  15. 15.

    et al. Analysis of the role of microsomal triglyceride transfer protein in the liver of tissue-specific knockout mice. J. Clin. Invest. 103, 1287–1298 (1999)

  16. 16.

    et al. Intestinal heat shock protein 110 regulates expression of CD1d on intestinal epithelial cells. J. Clin. Invest. 112, 745–754 (2003)

  17. 17.

    et al. Multiple defects in antigen presentation and T cell development by mice expressing cytoplasmic tail-truncated CD1d. Nature Immunol. 3, 55–60 (2002)

  18. 18.

    & The regulation of IL-10 production by immune cells. Nature Rev. Immunol. 10, 170–181 (2010)

  19. 19.

    et al. STAT3 links IL-22 signaling in intestinal epithelial cells to mucosal wound healing. J. Exp. Med. 206, 1465–1472 (2009)

  20. 20.

    , , & Hsp105β upregulates hsp70 gene expression through signal transducer and activator of transcription-3. FEBS J. 276, 5870–5880 (2009)

  21. 21.

    et al. Interleukin-10 signaling in regulatory T cells is required for suppression of Th17 cell-mediated inflammation. Immunity 34, 566–578 (2011)

  22. 22.

    et al. Regulatory T cell-derived interleukin-10 limits inflammation at environmental interfaces. Immunity 28, 546–558 (2008)

  23. 23.

    et al. Targeted disruption of Hsp110/105 gene protects against ischemic stress. Stroke 39, 2853–2859 (2008)

  24. 24.

    , & Immunoglobulin E production in the absence of interleukin-4-secreting CD1-dependent cells. Science 275, 977–979 (1997)

  25. 25.

    , , , & Interleukin-10-deficient mice develop chronic enterocolitis. Cell 75, 263–274 (1993)

  26. 26.

    et al. T cell-specific inactivation of the interleukin 10 gene in mice results in enhanced T cell responses but normal innate responses to lipopolysaccharide or skin irritation. J. Exp. Med. 200, 1289–1297 (2004)

  27. 27.

    et al. RAG-1-deficient mice have no mature B and T lymphocytes. Cell 68, 869–877 (1992)

  28. 28.

    , & Tissue-specific recognition of mouse CD1 molecules. J. Immunol. 160, 3128–3134 (1998)

  29. 29.

    et al. Hepatitis B virus-induced lipid alterations contribute to natural killer T cell-dependent protective immunity. Nature Med. 18, 1060–1068 (2012)

  30. 30.

    , , , & Immortalization of mouse intestinal epithelial cells by the SV40-large T gene. Phenotypic and immune characterization of the MODE-K cell line. J. Immunol. Methods 166, 63–73 (1993)

  31. 31.

    , , , & Diverse TCRs recognize murine CD1. J. Immunol. 162, 161–167 (1999)

  32. 32.

    , , , & CD1d ligation on human monocytes directly signals rapid NF-κB activation and production of bioactive IL-12. Proc. Natl Acad. Sci. USA 102, 11811–11816 (2005)

  33. 33.

    et al. A novel PPARγ gene therapy to control inflammation associated with inflammatory bowel disease in a murine model. Gastroenterology 124, 1315–1324 (2003)

  34. 34.

    & A simple method for constructing E1- and E1/E4-deleted recombinant adenoviral vectors. Hum. Gene Ther. 10, 2013–2017 (1999)

  35. 35.

    , , , & Carcinoembryonic antigen-related cell adhesion molecule 1 inhibits proximal TCR signaling by targeting ZAP-70. J. Immunol. 180, 6085–6093 (2008)

  36. 36.

    et al. XBP1 links ER stress to intestinal inflammation and confers genetic risk for human inflammatory bowel disease. Cell 134, 743–756 (2008)

  37. 37.

    et al. Eosinophils alter colonic epithelial barrier function: role for major basic protein. Am. J. Physiol. Gastrointest. Liver Physiol. 289, G890–G897 (2005)

  38. 38.

    et al. Hypoxia-inducible factor 1-dependent induction of intestinal trefoil factor protects barrier function during hypoxia. J. Exp. Med. 193, 1027–1034 (2001)

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Acknowledgements

The authors thank H.-C. Hung for technical assistance with microinjection, Y. Xie for performing osmium staining, A. Bedynek and M. Friedrich for performing immunohistochemistry of the human biopsies, F. A. Zhu for assistance with antigen presentation assays, D. Shouval, M. Sablon and D. Perez for animal care and husbandry, K. Tashiro for technical assistance with adenovirus preparation, V. M. Thiele for technical assistance, J. Cusick for help with manuscript preparation, and S. E. Plevy for discussions and reagents. This work was supported by: National Institutes of Health (NIH) (grants DK044319, DK051362, DK053056, DK088199) and the Harvard Digestive Diseases Center (DK0034854) (R.S.B.); the European Research Council (ERC Starting Grant agreement no. 336528), the Deutsche Forschungsgemeinschaft (DFG) (ZE 814/4-1, ZE 814/5-1, ZE 814/6-1), the Crohn’s and Colitis Foundation of America (Postdoctoral Fellowship Award), the European Commission (Marie Curie International Reintegration Grant no. 256363) and the DFG Excellence Cluster “Inflammation at Interfaces” (S.Z.); the DFG (OL 324/1-1) (T.O.); HL38180, DK56260, Washington University DDRCC P30DK52574 (morphology core) (N.O.D.); HDDC Pilot and Feasibility Grant (K.B.); NCI P30CA013696 (C.-S.L.), the DFG (BR 1912/6-1) and the Else Kroener-Fresenius-Stiftung (Else Kroener-Exzellenzstipendium 2010_EKES.32) (S.B.); Grant-in-Aid for Challenging Exploratory Research 24659823 from Japan Society for Promotion of Science (K.W.); the ERC under the European Community’s Seventh Framework Programme (FP7/2007-2013/ERC Grant agreement no. 260961), the National Institute for Health Research Cambridge Biomedical Research Centre, the Austrian Science Fund and Ministry of Science P21530-B18 and START Y446-B18, Innsbruck Medical University (MFI 2007-407) and the Addenbrooke’s Charitable Trust, CiCRA (A.K.); the European Community’s Seventh Framework Programme (FP7/2007-2013) under grant agreement SysmedIBD (no. 305564) (W.M., S.S.); the NIH (grants HL59561, DK034854, AI50950), the Helmsley Charitable Trust and the Wolpow Family Chair in IBD Treatment and Research (S.B.S.). PBS57-loaded and unloaded mouse CD1d tetramer was obtained through the NIH Tetramer Facility. The authors thank M. A. Exley and S. P. Colgan for discussions.

Author information

Author notes

    • Torsten Olszak
    • , Joana F. Neves
    • , C. Marie Dowds
    • , Sebastian Zeissig
    •  & Richard S. Blumberg

    These authors contributed equally to this work.

Affiliations

  1. Division of Gastroenterology, Hepatology, and Endoscopy, Brigham and Women’s Hospital, Harvard Medical School, Boston, Massachusetts 02115, USA

    • Torsten Olszak
    • , Joana F. Neves
    • , Kristi Baker
    • , Scott B. Snapper
    • , Sebastian Zeissig
    •  & Richard S. Blumberg
  2. Department of Internal Medicine I, University Medical Center Schleswig-Holstein, 24105 Kiel, Germany

    • C. Marie Dowds
    • , Stefan Schreiber
    •  & Sebastian Zeissig
  3. GI Pathology, Miraca Life Sciences, Newton, Massachusetts 02464, USA

    • Jonathan Glickman
  4. Division of Gastroenterology, Washington University School of Medicine, St Louis, Missouri 63110, USA

    • Nicholas O. Davidson
  5. Herbert Irving Comprehensive Cancer Center, Columbia University, New York, New York 10032, USA

    • Chyuan-Sheng Lin
  6. Department of Medicine, Department of Infectious Diseases & Pathology, University of Florida, Gainesville, Florida 32611, USA

    • Christian Jobin
  7. Department of Medicine II-Grosshadern, Ludwig Maximilians University, Munich 81377, Germany

    • Stephan Brand
  8. Institute of Pathology, Ludwig Maximilians University, Munich 80337, Germany

    • Karl Sotlar
  9. Department of Pharmacology, Graduate School of Dentistry, Osaka University, Osaka 565-0871, Japan

    • Koichiro Wada
    •  & Kazufumi Katayama
  10. Gastroenterology Division, Yokohama City University School of Medicine, Yokohama, Kanagawa 236-0027, Japan

    • Atsushi Nakajima
  11. Laboratory of Biochemistry and Molecular Biology, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka 565-0871, Japan

    • Hiroyuki Mizuguchi
  12. Department of Molecular Biosciences, Faculty of Life Sciences, Kyoto Sangyo University, Motoyama, Kamigamo, Kita-ku, Kyoto 603-8555, Japan

    • Kunito Kawasaki
    •  & Kazuhiro Nagata
  13. Faculty of Life Sciences, University of Manchester, Manchester M13 9PL, UK

    • Werner Müller
  14. Division of Pediatric Gastroenterology, Hepatology, and Nutrition, Department of Medicine, Children's Hospital Boston, Boston, Massachusetts 02115, USA

    • Scott B. Snapper
  15. Division of Gastroenterology, Addenbrooke Hospital, University of Cambridge, Cambridge CB2 0QQ, UK

    • Arthur Kaser

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Contributions

T.O., J.F.N., C.M.D. and K.B. performed in vitro and in vivo experiments and analysed the results; N.O.D. performed osmium tetroxide staining; J.G. obtained and scored histopathologies; C.-S.L. generated Cd1d1fl/fl mice; C.J. contributed to the analysis of CD1dΔIEC mice; S.B. and K.S. contributed to the immunohistochemical analysis of ulcerative colitis patients; K.W., K. Katayama, A.N. and H.M. generated adenoviruses; K. Kawasaki and K.N. provided HSP110-KO mice; W.M. and S.B.S. provided and participated in the analysis of the Il10ΔIEC mice; S.S. contributed to the coordination of experimental studies; A.K. contributed to MttpΔIEC studies and to the analysis of microarray data; R.S.B. and S.Z. designed the study, coordinated the experimental work and wrote the manuscript with input from co-authors. All authors discussed the results and commented on the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Sebastian Zeissig or Richard S. Blumberg.

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    Supplementary Information

    This file contains a Supplementary Discussion of potential mechanisms responsible for divergent outcomes of CD1d engagement on intestinal epithelial cells and professional antigen presenting cells.

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DOI

https://doi.org/10.1038/nature13150

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