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A two-amino-acid substitution in the transcription factor RORγt disrupts its function in TH17 differentiation but not in thymocyte development

Nature Immunology volume 18, pages 11281138 (2017) | Download Citation

Abstract

The transcription factor RORγt regulates differentiation of the TH17 subset of helper T cells, thymic T cell development and lymph-node genesis. Although elimination of RORγt prevents TH17 cell–mediated experimental autoimmune encephalomyelitis (EAE), it also disrupts thymocyte development, which could lead to lethal thymic lymphoma. Here we identified a two-amino-acid substitution in RORγt (RORγtM) that 'preferentially' disrupted TH17 differentiation but not thymocyte development. Mice expressing RORγtM were resistant to EAE associated with defective TH17 differentiation but maintained normal thymocyte development and normal lymph-node genesis, except for Peyer's patches. RORγtM showed less ubiquitination at Lys69 that was selectively required for TH17 differentiation but not T cell development. This study will inform the development of treatments that selectively target TH17 cell–mediated autoimmunity but do not affect thymocyte development or induce lymphoma.

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Acknowledgements

We thank J. C. Zuniga-Pflucker (University of Toronto) for the DP9-DL4 stroma cell line; W.S. Pear (University of Pennsylvania) for the retroviral vector MIGR; T. Dawson (Johns Hopkins) for RK5-HA-ubiquitin constructs; and Biocytogen for assisting with design and generation of the RorγtM/M mice. Supported by the US National Institutes of Health (R01-AI053147 and R01-AI109644), institutional pilot funding, the National Cancer Institute of the National Institutes of Health (P30CA33572, which includes work performed in the Animal Resource Center, Integrative Genomics, Analytical Cytometry, and Mass Spectrometry and Proteomics Cores) and the Science and Technology Department of Guangdong Province (2016A050503023 to Z. Huang). The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.

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Author notes

    • Zhiheng He
    •  & Jian Ma

    These authors contributed equally to this work.

Affiliations

  1. Division of Molecular Immunology, Beckman Research Institute of City of Hope, Duarte, California, USA.

    • Zhiheng He
    • , Jian Ma
    • , Ruiqing Wang
    • , Jing Zhang
    • , Fei Wang
    • , Subha Sen
    •  & Zuoming Sun
  2. Irell & Manella Graduate School of Biological Sciences, City of Hope, Duarte, California, USA.

    • Ruiqing Wang
    •  & Jing Zhang
  3. Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, Guangdong, P.R. China.

    • Zhaofeng Huang
  4. Division of Biology & Biological Engineering, California Institute of Technology, Pasadena, California, USA.

    • Ellen V Rothenberg

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Contributions

Z. He and Z.S. conceived of the study and wrote the manuscript; Z. He, J.M., R.W., J.Z., Z. Huang, F.W. and S.S. carried out the experiments. E.V.R. instructed and provided critical reagents for in vitro thymocyte development experiments; and all the authors reviewed and edited the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Zuoming Sun.

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https://doi.org/10.1038/ni.3832

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