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An alternative NFAT-activation pathway mediated by IL-7 is critical for early thymocyte development

Nature Immunology volume 14, pages 127–135 (2013)Cite this article

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Abstract

Interleukin 7 (IL-7) has a critical role in the development of early CD4−CD8− double-negative (DN) thymocytes. Although the transcription factor STAT5 is an important component of IL-7 signaling, differences in the phenotypes of mice deficient in STAT5, IL-7, IL-7 receptor alpha (IL-7rα) or the kinase Jak3 suggest the existence of STAT5-independent IL-7 signaling. Here we found that IL-7–Jak3 signals activated the transcription factor NFATc1 in DN thymocytes by phosphorylating Tyr371 in the regulatory region of NFATc1. This NFAT-activation pathway was critical for the survival and development of DN thymocytes, as deficiency in NFATc1 blocked thymocyte development at the DN1 stage, leading to T cell lymphopenia. In addition, our results demonstrated a cooperative function for NFATc1 and STAT5 in guiding thymocyte development in response to IL-7 signals.

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Figure 1: NFAT activation in DN thymocytes.
Figure 2: IL-7-induced activation of NFAT is independent of calcineurin activity.
Figure 3: IL-7-induced activation of NFATc1 is mediated by Jak3.
Figure 4: Jak3-induced activation of NFATc1 and translocation of NFATc1 to the nucleus is mediated by Tyr371 phosphorylation.
Figure 5: NFATc1 regulates Bcl-2 expression in early DN thymocytes.
Figure 6: Activation of NFAT is essential for the survival and differentiation of DN1–DN3 cells.
Figure 7: Loss of NFATc1 blocks T cell development at DN1 stage.

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Acknowledgements

We thank C. Linden for cell sorting; S.Y. Na, B. Kneitz and T. Hünig (University of Würzburg) for Rag1−/− and Il2−/− mice; A. Rao (La Jolla Institute for Allergy and Immunology) for Nfatc1fl/fl mice; D. Kioussis (Medical Research Council National Institute for Medical Research, London) for Vav-Cre mice; S. Chuvpilo (TheraMAB LLC, Moscow) for GST-NFATc1 constructs; R. Rost (Institute of Molecular Biology and Biochemistry, Graz, Austria) for NFATc1 expression vector; W.J. Leonard (US National Institutes of Health) for expression vectors for human IL-2Rγ and mouse IL-7Rα; J.J. O'Shea (US National Institutes of Health) for expression vectors for human Jak3; K. Hasegawa (Kyoto Medical Center) for the BCL2 luciferase construct; J.C. Zũniga-Pflücker (University of Toronto) for OP9 and OP9-DL1 cells; J. Schneider-Schaulies (University of Würzburg) for help in lentiviral transduction experiments; and members of the Serfling laboratory for comments on the manuscript. Supported by Deutsche Forschungsgemeinschaft (TRR52 to A.A. and E.S., and DFG-SFB854-TP9 to U.B.), the Wilhelm-Sander Foundation (E.S.) and the Mildred Scheel Foundation (E.S.).

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Authors and Affiliations

  1. Department of Molecular Pathology, Institute of Pathology, University of Würzburg, Würzburg, Germany

    Amiya K Patra, Andris Avots & Edgar Serfling

  2. Department of Bioanalytics, Leibniz-Institute for Analytical Sciences, Dortmund, Germany

    René P Zahedi & Albert Sickmann

  3. Institute of Molecular and Clinical Immunology, Otto-von-Guericke University Magdeburg, Magdeburg, Germany

    Thomas Schüler & Ursula Bommhardt

  4. Medical Proteom-Center, Ruhr-University Bochum, Bochum, Germany

    Albert Sickmann

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Contributions

A.K.P. conceived of the project and designed and did the experiments; A.A. assisted in OP9-DL1 coculture experiments and GST-protein purification; R.P.Z. and A.S. did mass spectrometry analysis for mapping of phosphorylated tyrosines; T.S. helped with Il7r−/− mice; U.B. contributed to immunofluorescence staining and discussions; E.S. contributed to the study organization; and A.K.P. and E.S. wrote the manuscript.

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Correspondence to Amiya K Patra or Edgar Serfling.

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Patra, A., Avots, A., Zahedi, R. et al. An alternative NFAT-activation pathway mediated by IL-7 is critical for early thymocyte development. Nat Immunol 14, 127–135 (2013). https://doi.org/10.1038/ni.2507

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