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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 658666 (2008) | Download Citation

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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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.).

Author information

Author notes

    • Jeff Holst

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

    • Haopeng Wang
    • , Kelly Durick Eder
    •  & Creg J Workman

    These authors contributed equally to this work.

Affiliations

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

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

    • Zachary Baquet
    •  & Richard Smeyne
  4. Interdisciplinary Program, University of Tennessee Health Science Center, Memphis, Tennessee 38163, USA.

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

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

    • Harvir Singh
    • , Andrzej Chruscinski
    •  & Paul J Utz
  7. Department of Immunology and Department of Microbiology, The University of Texas Southwestern Medical Center, Dallas, Texas 75390-9093, 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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DOI

https://doi.org/10.1038/ni.1611

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