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A genetically selective inhibitor demonstrates a function for the kinase Zap70 in regulatory T cells independent of its catalytic activity

Nature Immunology volume 11, pages 1085–1092 (2010)Cite this article

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Abstract

To investigate the role of the kinase Zap70 in T cells, we generated mice expressing a Zap70 mutant whose catalytic activity can be selectively blocked by a small-molecule inhibitor. We found that conventional naive, effector and memory T cells were dependent on the kinase activity of Zap70 for their activation, which demonstrated a nonredundant role for Zap70 in signals induced by the T cell antigen receptor (TCR). In contrast, the catalytic activity of Zap70 was not required for activation of the GTPase Rap1 and inside-out signals that promote integrin adhesion. This Zap70 kinase–independent pathway was sufficient for the suppressive activity of regulatory T cells (Treg cells), which was unperturbed by inhibition of the catalytic activity of Zap70. Our results indicate Zap70 is a likely therapeutic target.

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Figure 1: T cell activation is dependent on the catalytic activity of Zap70.
Figure 2: Proliferation of CD4+ T cells requires the catalytic activity of Zap70.
Figure 3: Execution of effector T cell functions requires the catalytic activity of Zap70.
Figure 4: CD8+ memory responses are dependent on the catalytic activity of Zap70.
Figure 5: The catalytic activity of Zap70 is dispensable for the suppressive activity of Treg cells in vitro.
Figure 6: TCR-induced activation of Rap1 and adhesion to ICAM-1 are independent of Zap70 kinase activity.
Figure 7: TCR-induced activation of Rap1 and adhesion to ICAM-1 are dependent on the adaptor function of Zap70.

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Change history

  • 04 November 2010

    In the version of this article initially published online, the key above Figure 5a was mislabeled. The correct label is Treg:Tconv. The error has been corrected for the print, PDF and HTML versions of this article.

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Acknowledgements

We thank M. Gately (Hoffmann–La Roche) for recombinant human IL-2; A. Roque for assistance in animal husbandry; C. MacArthur for cell sorting; and J. Wei for generating the Zap70(AS) BAC transgene construct. Supported by the National Institute of Arthritis, Musculoskeletal and Skin Diseases, National Institutes of Health (RC2AR058947 to A.W. and F32AR056174 to L.-Y.H.).

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  1. Susan E Levin

    Present address: Present address: Department of Biology, Tufts University, Medford, Massachusetts.,

Authors and Affiliations

  1. Department of Medicine, Department of Microbiology and Immunology, Howard Hughes Medical Institute, Rosalind Russell Medical Research Center for Arthritis, University of California, San Francisco, San Francisco, California, USA

    Byron B Au-Yeung, Susan E Levin, Lih-Yun Hsu, Debra A Cheng & Arthur Weiss

  2. Department of Cellular and Molecular Pharmacology, Howard Hughes Medical Institute, University of California, San Francisco, San Francisco, California, USA

    Chao Zhang & Kevan M Shokat

  3. Department of Microbiology and Immunology, University of California, San Francisco, San Francisco, California, USA

    Nigel Killeen

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Contributions

B.B.A.-Y. did most of the experiments and wrote the paper; S.E.L. did the initial characterization of Zap70(AS) mice; N.K. designed the strategy for generating Zap70(AS) mice; D.A.C. assisted with the ICAM-1 adhesion assays and calcium-flux assays; L.-Y.H. helped with the Zap70(YYAA) Treg cell–suppression assay; K.M.S. and C.Z. provided advice and synthesized 3-MB-PP1; and A.W. directed the project.

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Correspondence to Arthur Weiss.

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The authors declare no competing financial interests.

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Au-Yeung, B., Levin, S., Zhang, C. et al. A genetically selective inhibitor demonstrates a function for the kinase Zap70 in regulatory T cells independent of its catalytic activity. Nat Immunol 11, 1085–1092 (2010). https://doi.org/10.1038/ni.1955

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