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Mutation of Tec family kinases alters T helper cell differentiation

Nature Immunology volume 2, pages 1183–1188 (2001)Cite this article

Abstract

The Tec kinases Rlk and Itk are critical for full T cell receptor (TCR)-induced activation of phospholipase C-γ and mitogen-activated protein kinase. We show here that the mutation of Rlk and Itk impaired activation of the transcription factors NFAT and AP-1 and production of both T helper type 1 (TH1) and TH2 cytokines. Consistent with these biochemical defects, Itk−/− mice did not generate effective TH2 responses when challenged with Schistosoma mansoni eggs. Paradoxically, the more severely impaired Rlk−/−Itk−/− mice were able to mount a TH2 response and produced TH2 cytokines in response to this challenge. In addition, Rlk−/−Itk−/− cells showed impaired TCR-induced repression of the TH2-inducing transcription factor GATA-3, suggesting a potential mechanism for TH2 development in these hyporesponsive cells. Thus, mutations that affect Tec kinases lead to complex alterations in CD4+ TH cell differentiation.

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Figure 1: Decreased activation of NFATc and NFATp in Rlk−/−Itk−/− T lymphocytes.
Figure 2: Decreased TCR-mediated responses in CD4+ Rlk−/−Itk−/− T cells.
Figure 3: Increased IgG1 and IgE in Rlk−/−Itk−/− and Itk−/− mice.
Figure 4: Altered responses to the TH2 polarizing agent S. mansoni.
Figure 5: Itk−/− and Rlk−/−Itk−/− CD4+ T cells polarize in culture but have decreased cytokine production.
Figure 6: Impaired TCR-induced GATA-3 repression in Rlk−/−Itk−/− cells.

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Acknowledgements

We thank M. Erdos, S. John, U. Siebenlist and K. Brown for advice on EMSAs; J. Hu-Li and D. Jankovic for advice on in vitro polarization cultures; J. Cote-Sierra for advice on GATA-3 assays; A. Venegas, M. Chetana, J. Shehadeh and G. Elliott for technical assistance; S. Barbieri, R. Swafford, and C. Eigsti for cell sorting; A. Rao, J. Powell and N. Rice for gifts of antibodies; and R. Germain, D. Jankovic, T. Wynn, M. Kullberg, J. Bluestone, W. Paul and A. Kimmel for critical discussions. Supported in part by the Searle Scholars Program/Chicago Community Trust (P. L. S.) and the HHMI-NIH Scholars Program (E. M. S. and C. M. L.).

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Author notes

  1. Edward M. Schaeffer, George S. Yap and Carol M. Lewis: These authors contributed equally to this work.

Authors and Affiliations

  1. National Human Genome Research Institute, National Institutes of Health, Bethesda, 20892, MD, USA

    Edward M. Schaeffer, Carol M. Lewis, Michael J. Czar & Pamela L. Schwartzberg

  2. National Institute of Allergy and Infectious Disease and National Institutes of Health, Bethesda, MD, USA

    George S. Yap & Alan Sher

  3. National Cancer Institute, National Institutes of Health, Bethesda, MD, USA

    Daniel W. McVicar

  4. Biomedical Research Institute, Rockville, MD, USA

    Allen W. Cheever

  5. Department of Medical Microbiology and Immunology, Brown University, Providence, RI, USA

    George S. Yap

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Correspondence to Pamela L. Schwartzberg.

Supplementary information

Web Figure 1

GATA-3 protein expression. (a) Purified T cells were stimulated with anti-CD3 for 40 h in the absence of cytokines, nuclear extracts prepared and GATA-3 protein expression examined by immunoblotting with monoclonal anti-GATA-3 or goat polyclonal anti-STAT6. (b) The specificity of the GATA-3 antibody was examined in nuclear extracts from cells that had been differentiated with TH1 or TH2 cytokines for 40 h. (JPG 23 kb)

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Schaeffer, E., Yap, G., Lewis, C. et al. Mutation of Tec family kinases alters T helper cell differentiation. Nat Immunol 2, 1183–1188 (2001). https://doi.org/10.1038/ni734

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