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Scalable signaling mediated by T cell antigen receptor–CD3 ITAMs ensures effective negative selection and prevents autoimmunity

Nature Immunology volume 9, pages 658–666 (2008)Cite this article

Abstract

The T cell antigen receptor (TCR)-CD3 complex is unique in having ten cytoplasmic immunoreceptor tyrosine-based activation motifs (ITAMs). The physiological importance of this high TCR ITAM number is unclear. Here we generated 25 groups of mice expressing various combinations of wild-type and mutant ITAMs in TCR-CD3 complexes. Mice with fewer than seven wild-type CD3 ITAMs developed a lethal, multiorgan autoimmune disease caused by a breakdown in central rather than peripheral tolerance. Although there was a linear correlation between the number of wild-type CD3 ITAMs and T cell proliferation, cytokine production was unaffected by ITAM number. Thus, high ITAM number provides scalable signaling that can modulate proliferation yet ensure effective negative selection and prevention of autoimmunity.

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Figure 1: Profound autoimmunity in CD3-mutant mice.
Figure 2: A specific number of ITAMs is required for each T cell developmental parameter.
Figure 3: Cells from CD3-mutant mice proliferate less but have normal cytokine production.
Figure 4: Normal proportion and function of Treg cells in CD3-mutant mice.
Figure 5: Lower signaling converts negative selection into positive selection and generates IFN-γ-producing autoreactive T cells.

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Acknowledgements

We thank D. Green, M. Davis and J. Ihle for reviewing the manuscript; L. Collison (St. Jude Children's Research Hospital) for mice with inflammatory bowel disease; C. Terhorst (Harvard Medical School) for Cd3eΔP/ΔP mice; N. van Oers (University of Texas Southwestern Medical Center) for CD3ζ-KO Tg1-6M mice; M. Davis for 3A9 TCR–transgenic mice; K. Vignali, Y. Wang, S. Dilioglou, A. Burton and E. Vincent for technical assistance; the Vignali lab for assistance with bone marrow collection; R. Cross, J. Hoffrage, J. Smith and Y. He for flow cytometry; S. Rowe, J. Gatewood and J. Smith for cytokine analysis; L. Zhang of the Cell and Tissue Imaging facility for image acquisition; the staff of the Flow Cytometry and Cell Sorting Shared Resource facility for purification by magnetic-activated cell sorting; and the Hartwell Center for DNA sequencing. Supported by the National Institutes of Health (AI-52199; and U19-DK-6134 to P.J.U.), the St. Jude National Cancer Institute Cancer Center (CA-21765) and the American Lebanese Syrian Associated Charities (D.A.A.V.) the National Heart, Lung and Blood Institute (proteomics contract NOI-HV-28183 to P.J.U.); and the Floren Family Trust (P.J.U.).

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Author notes

  1. Jeff Holst

    Present address: Present address: Gene & Stem Cell Therapy Program, Centenary Institute of Cancer Medicine & Cell Biology, University of Sydney, NSW 2042, Australia.,

  2. Haopeng Wang, Kelly Durick Eder and Creg J Workman: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Immunology, St. Jude Children's Research Hospital, Memphis, 38105-2794, Tennessee, USA

    Jeff Holst, Haopeng Wang, Kelly Durick Eder, Creg J Workman, Karen Forbes & Dario A A Vignali

  2. Animal Resource Center, St. Jude Children's Research Hospital, Memphis, 38105-2794, Tennessee, USA

    Kelli L Boyd

  3. Department of Developmental Neurobiology, St. Jude Children's Research Hospital, Memphis, 38105-2794, Tennessee, USA

    Zachary Baquet & Richard Smeyne

  4. Interdisciplinary Program, University of Tennessee Health Science Center, Memphis, 38163, Tennessee, USA

    Haopeng Wang

  5. Department of Pathology, University of Tennessee Health Science Center, Memphis, 38163, Tennessee, USA

    Dario A A Vignali

  6. Division of Immunology & Rheumatology, Department of Medicine, Stanford University School of Medicine, Stanford, 94305, California, USA

    Harvir Singh, Andrzej Chruscinski & Paul J Utz

  7. Department of Immunology and Department of Microbiology, The University of Texas Southwestern Medical Center, Dallas, 75390-9093, Texas, USA

    Nicolai S C van Oers

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Contributions

All authors contributed to discussions of experimental design, data analysis and/or editing of the manuscript; J.H. did all experiments unless stated otherwise; H.W., K.D.E. and C.J.W. generated mice for the immunofluorescence microscopy and autoantibody analysis, did the Treg cell experiments, chimera-lymphopenia experiments, some of the flow cytometry analysis and the BrdU and lymph node assays; K.L.B. did all the histological analysis and scoring while 'blinded' to sample identity; K.F. assisted in cellular isolation and analysis and managed the breeding colonies; Z.B. and R.S. did the immunofluorescence microscopy, H.S., A.C. and P.J.U. did the autoantibody analysis; N.S.C.v.O. provided the CD3ζ-KO Tg1-6M-transgenic mice and discussions about their use; J.H. and D.A.A.V. wrote the manuscript; and D.A.A.V. conceptualized and directed the project.

Corresponding author

Correspondence to Dario A A Vignali.

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Holst, J., Wang, H., Eder, K. et al. Scalable signaling mediated by T cell antigen receptor–CD3 ITAMs ensures effective negative selection and prevents autoimmunity. Nat Immunol 9, 658–666 (2008). https://doi.org/10.1038/ni.1611

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