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  • A Corrigendum to this article was published on 16 March 2016

Abstract

T-cell immunoglobulin domain and mucin domain-3 (TIM-3, also known as HAVCR2) is an activation-induced inhibitory molecule involved in tolerance and shown to induce T-cell exhaustion in chronic viral infection and cancers1,2,3,4,5. Under some conditions, TIM-3 expression has also been shown to be stimulatory. Considering that TIM-3, like cytotoxic T lymphocyte antigen 4 (CTLA-4) and programmed death 1 (PD-1), is being targeted for cancer immunotherapy, it is important to identify the circumstances under which TIM-3 can inhibit and activate T-cell responses. Here we show that TIM-3 is co-expressed and forms a heterodimer with carcinoembryonic antigen cell adhesion molecule 1 (CEACAM1), another well-known molecule expressed on activated T cells and involved in T-cell inhibition6,7,8,9,10. Biochemical, biophysical and X-ray crystallography studies show that the membrane-distal immunoglobulin-variable (IgV)-like amino-terminal domain of each is crucial to these interactions. The presence of CEACAM1 endows TIM-3 with inhibitory function. CEACAM1 facilitates the maturation and cell surface expression of TIM-3 by forming a heterodimeric interaction in cis through the highly related membrane-distal N-terminal domains of each molecule. CEACAM1 and TIM-3 also bind in trans through their N-terminal domains. Both cis and trans interactions between CEACAM1 and TIM-3 determine the tolerance-inducing function of TIM-3. In a mouse adoptive transfer colitis model, CEACAM1-deficient T cells are hyper-inflammatory with reduced cell surface expression of TIM-3 and regulatory cytokines, and this is restored by T-cell-specific CEACAM1 expression. During chronic viral infection and in a tumour environment, CEACAM1 and TIM-3 mark exhausted T cells. Co-blockade of CEACAM1 and TIM-3 leads to enhancement of anti-tumour immune responses with improved elimination of tumours in mouse colorectal cancer models. Thus, CEACAM1 serves as a heterophilic ligand for TIM-3 that is required for its ability to mediate T-cell inhibition, and this interaction has a crucial role in regulating autoimmunity and anti-tumour immunity.

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Acknowledgements

We thank T. Gallagher, M. Yoshida and K. Holmes for essential reagents, R. Gali for statistical assistance, C. Chen, T. Wesse, S. Sabet, S. Greve, T. Henke, D. Tan, K. Sakuishi and J. Sullivan for technical assistance, E. Greenfield and C. Bencsics for core services, and J. H. Wang, E. Reinherz, R. Grenha, H. Iijima, J. Shively, A. Kaser, T. E. Adolph, K. Baker, D. Ringe and S. Zeissig for discussions. We thank the staff of the Dana Farber/Harvard Cancer Center monoclonal antibody core for purification of proteins used in X-ray crystallography and beam line X25 and X6A of the National Synchrotron Light Source (NSLS), Brookhaven National Laboratory, USA. The NSLS is supported by the US Department of Energy. This work was supported by the American Cancer Society grant RSG-11-057-01-LIB (A.C.A.); the Norwegian PSC research center and the Unger Vetlesen Medical Fund (E.M.); Crohn’s & Colitis Foundation of America fellowship grant (Y.-H.H.); Deutsche Forschungsgemeinschaft (DFG) Cluster of Excellence ‘Inflammation at Interfaces’ Award (A.F. and B.-S.P.); Harvard Clinical Translational Science Center, UL1 TR001102 (R. Gali); the National Basic Research Program of China No. 2010CB529906 (Q.C.); Canadian Institute of Health Research (K.L.C. and N.B.); Canadian Institute of Health Research grant MOP-93787 (M.A.O.); AACR-Pancreatic Cancer Action Network (H.L.P. and S.K.D.); National Institutes of Health (NIH) grant GM32415 (G.A.P.); NIH grants AI073748, NS045937, AI039671 and AI056299 (V.K.K.); NIH grants DK044319, DK051362, DK053056, DK088199, the Harvard Digestive Diseases Center (HDDC) DK0034854 and High Point Foundation (R.S.B.).

Author information

Author notes

    • Chen Zhu
    •  & Yasuyuki Kondo

    These authors contributed equally to this work.

Affiliations

  1. Division of Gastroenterology, Department of Medicine, Brigham and Women’s Hospital, Harvard Medical School, 75 Francis Street, Boston, Massachusetts 02115, USA

    • Yu-Hwa Huang
    • , Yasuyuki Kondo
    • , Amit Gandhi
    • , Espen Melum
    • , Michal Pyzik
    •  & Richard S. Blumberg
  2. Evergrande Center for Immunologic Diseases, Harvard Medical School and Brigham and Women’s Hospital, Harvard Institutes of Medicine, 77 Avenue Louis Pasteur, Boston, Massachusetts 02115, USA

    • Chen Zhu
    • , Ana C. Anderson
    • , Thomas Pertel
    •  & Vijay K. Kuchroo
  3. Rosenstiel Basic Medical Sciences Research Center, Brandeis University, 415 South Street, Waltham, Massachusetts 02454, USA

    • Andrew Russell
    •  & Gregory A. Petsko
  4. Whitehead Institute, Massachusetts Institute of Technology, Cambridge, Massachusetts 02142, USA

    • Stephanie K. Dougan
    •  & Hidde L. Ploegh
  5. Institute of Clinical Molecular Biology, Christian-Albrechts-University of Kiel, Kiel 24105, Germany

    • Britt-Sabina Petersen
    •  & Andre Franke
  6. Norwegian PSC Research Center, Division of Cancer Medicine, Surgery and Transplantation, Oslo University Hospital, Oslo 0424, Norway

    • Espen Melum
  7. Department of Immunology, University of Toronto, Toronto, Ontario M5S1A8, Canada

    • Kiera L. Clayton
    •  & Mario A. Ostrowski
  8. Cell Signalling Section, Department of Pathology, University of Cambridge, Cambridge CB2 1QP, UK

    • Monika Raab
    •  & Christopher E. Rudd
  9. State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu 610041, China

    • Qiang Chen
  10. Goodman Cancer Research Centre, McGill University, Montreal H3G 1Y6, Canada

    • Nicole Beauchemin
  11. Beckman Institute, City of Hope, Duarte, California 91010, USA

    • Paul J. Yazaki
  12. Keenan Research Centre of St. Michael’s Hospital, Toronto, Ontario M5S1A8, Canada

    • Mario A. Ostrowski
  13. GI Pathology, Miraca Life Sciences, Newton, Massachusetts 02464, USA

    • Jonathan N. Glickman

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Contributions

Y.-H.H., C.Z. and Y.K. performed most experiments and helped prepare the manuscript. B.-S.P., E.M. and A.F. provided expertise in the genetic assessment for TIM-3. J.N.G. assessed all pathology. A.C.A. designed and directed tumour experiments. T.P. designed shRNA experiments. M.R. and C.E.R. performed proximity ligation analysis. S.K.D. and H.L.P. conducted and analysed pulse-chase biosynthetic labelling experiments. A.G., A.R., Q.C. and G.A.P. performed X-ray crystallography or structural analysis. K.L.C. and M.A.O. conducted immune synapse experiments. M.P. and P.J.Y. assisted with the single chain protein analysis. N.B. assisted in generation of Ceacam1−/− Rag2−/− mice and in analyses of data. R.S.B. and V.K.K. devised and coordinated the project, and together with Y.-H.H., C.Z. and Y.K. wrote the manuscript and designed the experiments. R.S.B. and V.K.K. share senior authorship on this paper.

Competing interests

V.K.K. and A.C.A. are consultants to Novartis, which is developing immune modulators for the treatment of cancer; R.S.B. and all other authors declare no competing financial interests.

Corresponding author

Correspondence to Richard S. Blumberg.

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https://doi.org/10.1038/nature13848

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