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Abstract

CTLA-4, a negative regulator of T cell function, was found to associate with the T cell receptor (TCR) complex ζ chain in primary T cells. The association of TCRζ with CTLA-4, reconstituted in 293 transfectants, was enhanced by p56lck-induced tyrosine phosphorylation. Coexpression of the CTLA-4–associated tyrosine phosphatase, SHP-2, resulted in dephosphorylation of TCRζ bound to CTLA-4 and abolished the p56lck-inducible TCRζ–CTLA-4 interaction. Thus, CTLA-4 inhibits TCR signal transduction by binding to TCRζ and inhibiting tyrosine phosphorylation after T cell activation. These findings have broad implications for the negative regulation of T cell function and T cell tolerance.

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REFERENCES AND NOTES

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21
Immunoprecipitations were performed with lysates prepared in LB [1% Nonidet P-40, 50 mM tris-HCl (pH 7.4), 150 mM NaCl, 20 mM EDTA (pH 8.0), 1 mM sodium vanadate, leupeptin (10 μg/ml), 10 μM aprotinin, 1 mM phenylsulfonylsulfoxide]. Lysates were precleared twice with protein A beads and once with protein A beads coated with a mAb from a control hamster (UC3-10A6) (18) before precipitating antibodies were added. Immunoprecipitation was performed overnight at 4oC. Immune complexes were washed five times with LB and subjected to SDS-PAGE.
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Supported by the National Institutes of Health (PHS PO1 AI35294-6). Additional support was provided by a Howard Hughes Medical Institute Medical Student Research Training Fellowship (D.K.H.) and a Mayo Foundation Scholarship (M.G.). We thank G. Feng for the reagents; A. Sharpe for CTLA-4 knockout mice; N. Patel and C. Long for technical assistance; S. Thomas, M. Fonstein, and W. Buikema for services provided at the Cancer Research Center (CRC) DNA Sequencing Facility; and M. Clark for critical reading of the manuscript.

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Information & Authors

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Published In

Science
Volume 282 | Issue 5397
18 December 1998

Submission history

Received: 10 August 1998
Accepted: 10 November 1998
Published in print: 18 December 1998

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Authors

Affiliations

Kyung-Mi Lee*
K.-M. Lee, M. Griffin, R. Khattri, D. K. Hong, J. A. Bluestone, Ben May Institute for Cancer Research, and Committee on Immunology, University of Chicago, Chicago, IL 60637, USA. E. Chuang, Committee on Immunology, and Gwen Knapp Center for Lupus and Immunology Research, University of Chicago, Chicago, IL 60637, USA. W. Zhang and L. E. Samelson, Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA. D. Straus, Committee on Immunology, and Department of Pathology and Medicine, University of Chicago, Chicago, IL 60637, USA. C. B. Thompson, Howard Hughes Medical Institute, Ben May Institute for Cancer Research, Committee on Immunology, and Gwen Knapp Center for Lupus and Immunology Research, University of Chicago, Chicago, IL 60637, USA.
Ellen Chuang*
K.-M. Lee, M. Griffin, R. Khattri, D. K. Hong, J. A. Bluestone, Ben May Institute for Cancer Research, and Committee on Immunology, University of Chicago, Chicago, IL 60637, USA. E. Chuang, Committee on Immunology, and Gwen Knapp Center for Lupus and Immunology Research, University of Chicago, Chicago, IL 60637, USA. W. Zhang and L. E. Samelson, Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA. D. Straus, Committee on Immunology, and Department of Pathology and Medicine, University of Chicago, Chicago, IL 60637, USA. C. B. Thompson, Howard Hughes Medical Institute, Ben May Institute for Cancer Research, Committee on Immunology, and Gwen Knapp Center for Lupus and Immunology Research, University of Chicago, Chicago, IL 60637, USA.
Matthew Griffin
K.-M. Lee, M. Griffin, R. Khattri, D. K. Hong, J. A. Bluestone, Ben May Institute for Cancer Research, and Committee on Immunology, University of Chicago, Chicago, IL 60637, USA. E. Chuang, Committee on Immunology, and Gwen Knapp Center for Lupus and Immunology Research, University of Chicago, Chicago, IL 60637, USA. W. Zhang and L. E. Samelson, Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA. D. Straus, Committee on Immunology, and Department of Pathology and Medicine, University of Chicago, Chicago, IL 60637, USA. C. B. Thompson, Howard Hughes Medical Institute, Ben May Institute for Cancer Research, Committee on Immunology, and Gwen Knapp Center for Lupus and Immunology Research, University of Chicago, Chicago, IL 60637, USA.
Roli Khattri
K.-M. Lee, M. Griffin, R. Khattri, D. K. Hong, J. A. Bluestone, Ben May Institute for Cancer Research, and Committee on Immunology, University of Chicago, Chicago, IL 60637, USA. E. Chuang, Committee on Immunology, and Gwen Knapp Center for Lupus and Immunology Research, University of Chicago, Chicago, IL 60637, USA. W. Zhang and L. E. Samelson, Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA. D. Straus, Committee on Immunology, and Department of Pathology and Medicine, University of Chicago, Chicago, IL 60637, USA. C. B. Thompson, Howard Hughes Medical Institute, Ben May Institute for Cancer Research, Committee on Immunology, and Gwen Knapp Center for Lupus and Immunology Research, University of Chicago, Chicago, IL 60637, USA.
David K. Hong
K.-M. Lee, M. Griffin, R. Khattri, D. K. Hong, J. A. Bluestone, Ben May Institute for Cancer Research, and Committee on Immunology, University of Chicago, Chicago, IL 60637, USA. E. Chuang, Committee on Immunology, and Gwen Knapp Center for Lupus and Immunology Research, University of Chicago, Chicago, IL 60637, USA. W. Zhang and L. E. Samelson, Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA. D. Straus, Committee on Immunology, and Department of Pathology and Medicine, University of Chicago, Chicago, IL 60637, USA. C. B. Thompson, Howard Hughes Medical Institute, Ben May Institute for Cancer Research, Committee on Immunology, and Gwen Knapp Center for Lupus and Immunology Research, University of Chicago, Chicago, IL 60637, USA.
Weiguo Zhang
K.-M. Lee, M. Griffin, R. Khattri, D. K. Hong, J. A. Bluestone, Ben May Institute for Cancer Research, and Committee on Immunology, University of Chicago, Chicago, IL 60637, USA. E. Chuang, Committee on Immunology, and Gwen Knapp Center for Lupus and Immunology Research, University of Chicago, Chicago, IL 60637, USA. W. Zhang and L. E. Samelson, Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA. D. Straus, Committee on Immunology, and Department of Pathology and Medicine, University of Chicago, Chicago, IL 60637, USA. C. B. Thompson, Howard Hughes Medical Institute, Ben May Institute for Cancer Research, Committee on Immunology, and Gwen Knapp Center for Lupus and Immunology Research, University of Chicago, Chicago, IL 60637, USA.
David Straus
K.-M. Lee, M. Griffin, R. Khattri, D. K. Hong, J. A. Bluestone, Ben May Institute for Cancer Research, and Committee on Immunology, University of Chicago, Chicago, IL 60637, USA. E. Chuang, Committee on Immunology, and Gwen Knapp Center for Lupus and Immunology Research, University of Chicago, Chicago, IL 60637, USA. W. Zhang and L. E. Samelson, Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA. D. Straus, Committee on Immunology, and Department of Pathology and Medicine, University of Chicago, Chicago, IL 60637, USA. C. B. Thompson, Howard Hughes Medical Institute, Ben May Institute for Cancer Research, Committee on Immunology, and Gwen Knapp Center for Lupus and Immunology Research, University of Chicago, Chicago, IL 60637, USA.
Lawrence E. Samelson
K.-M. Lee, M. Griffin, R. Khattri, D. K. Hong, J. A. Bluestone, Ben May Institute for Cancer Research, and Committee on Immunology, University of Chicago, Chicago, IL 60637, USA. E. Chuang, Committee on Immunology, and Gwen Knapp Center for Lupus and Immunology Research, University of Chicago, Chicago, IL 60637, USA. W. Zhang and L. E. Samelson, Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA. D. Straus, Committee on Immunology, and Department of Pathology and Medicine, University of Chicago, Chicago, IL 60637, USA. C. B. Thompson, Howard Hughes Medical Institute, Ben May Institute for Cancer Research, Committee on Immunology, and Gwen Knapp Center for Lupus and Immunology Research, University of Chicago, Chicago, IL 60637, USA.
Craig B. Thompson
K.-M. Lee, M. Griffin, R. Khattri, D. K. Hong, J. A. Bluestone, Ben May Institute for Cancer Research, and Committee on Immunology, University of Chicago, Chicago, IL 60637, USA. E. Chuang, Committee on Immunology, and Gwen Knapp Center for Lupus and Immunology Research, University of Chicago, Chicago, IL 60637, USA. W. Zhang and L. E. Samelson, Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA. D. Straus, Committee on Immunology, and Department of Pathology and Medicine, University of Chicago, Chicago, IL 60637, USA. C. B. Thompson, Howard Hughes Medical Institute, Ben May Institute for Cancer Research, Committee on Immunology, and Gwen Knapp Center for Lupus and Immunology Research, University of Chicago, Chicago, IL 60637, USA.
Jeffrey A. Bluestone
K.-M. Lee, M. Griffin, R. Khattri, D. K. Hong, J. A. Bluestone, Ben May Institute for Cancer Research, and Committee on Immunology, University of Chicago, Chicago, IL 60637, USA. E. Chuang, Committee on Immunology, and Gwen Knapp Center for Lupus and Immunology Research, University of Chicago, Chicago, IL 60637, USA. W. Zhang and L. E. Samelson, Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA. D. Straus, Committee on Immunology, and Department of Pathology and Medicine, University of Chicago, Chicago, IL 60637, USA. C. B. Thompson, Howard Hughes Medical Institute, Ben May Institute for Cancer Research, Committee on Immunology, and Gwen Knapp Center for Lupus and Immunology Research, University of Chicago, Chicago, IL 60637, USA.

Notes

*
These authors contributed equally to this work.
To whom correspondence should be addressed. E-mail: [email protected]

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