Mutation of Tec family kinases alters T helper cell differentiation

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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Correspondence to Pamela L. Schwartzberg.

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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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