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Published October 2017 | Supplemental Material + Accepted Version
Journal Article Open

A two-amino-acid substitution in the transcription factor RORγt disrupts its function in T_H17 differentiation but not in thymocyte development

Abstract

The transcription factor RORγt regulates differentiation of the T_H17 subset of helper T cells, thymic T cell development and lymph-node genesis. Although elimination of RORγt prevents T_H17 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 (RORγt^M) that 'preferentially' disrupted T_H17 differentiation but not thymocyte development. Mice expressing RORγt^M were resistant to EAE associated with defective T_H17 differentiation but maintained normal thymocyte development and normal lymph-node genesis, except for Peyer's patches. RORγt^M showed less ubiquitination at Lys69 that was selectively required for T_H17 differentiation but not T cell development. This study will inform the development of treatments that selectively target T_H17 cell–mediated autoimmunity but do not affect thymocyte development or induce lymphoma.

Additional Information

© 2017 Macmillan Publishers Limited, part of Springer Nature. Received 11 April; accepted 8 August; published online 28 August 2017. 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 Rorγt^(M/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. Author 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. The authors declare no competing financial interests.

Attached Files

Accepted Version - nihms898687.pdf

Supplemental Material - ni.3832-S1.pdf

Supplemental Material - ni.3832-S2.pdf

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Additional details

Created:
August 19, 2023
Modified:
October 17, 2023