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TET proteins regulate the lineage specification and TCR-mediated expansion of iNKT cells

Nature Immunology volume 18, pages 4553 (2017) | Download Citation

Abstract

TET proteins oxidize 5-methylcytosine in DNA to 5-hydroxymethylcytosine and other oxidation products. We found that simultaneous deletion of Tet2 and Tet3 in mouse CD4+CD8+ double-positive thymocytes resulted in dysregulated development and proliferation of invariant natural killer T cells (iNKT cells). Tet2-Tet3 double-knockout (DKO) iNKT cells displayed pronounced skewing toward the NKT17 lineage, with increased DNA methylation and impaired expression of genes encoding the key lineage-specifying factors T-bet and ThPOK. Transfer of purified Tet2-Tet3 DKO iNKT cells into immunocompetent recipient mice resulted in an uncontrolled expansion that was dependent on the nonclassical major histocompatibility complex (MHC) protein CD1d, which presents lipid antigens to iNKT cells. Our data indicate that TET proteins regulate iNKT cell fate by ensuring their proper development and maturation and by suppressing aberrant proliferation mediated by the T cell antigen receptor (TCR).

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Acknowledgements

We thank C. Kim, K. van Gunst, L. Nosworthy, D. Hinz and R. Simmons at the LJI Flow Cytometry Core for help with fluorescence-activated cell sorting; G. Seumois and J. Day at the LJI Functional Genomics Center for assistance with next-generation sequencing (Illumina HiSeq 2500); M. Kronenberg, I. Engel and C.-W. Lio (LJI) for discussions, the LJI Bioinformatics Core for routine analysis; Z. Mikulski and B. Kiosses at the LJI microscopy core, M. Chadwell at the LJI Histology core, and the Histology Core at the University of California at San Diego Moores Cancer Center; and R. Bosselut (National Cancer Institute) for pMX-ThPOK-IRES-GFP. Supported by US National Institutes of Health (R01 AI44432, CA151535 and R35CA210043), the Leukemia and Lymphoma Society (Translation Research Project grant 6187-12 to A.R.), the Academy of Finland Centre of Excellence in Molecular Systems Immunology and Physiology Research (H.L. and. S.R.), an Albert Billings Ruddock Professorship at Caltech (E.V.R.), the Cancer Research Institute (Irvington Institute postdoctoral fellowship to A.T.), the Fraternal Order of Eagles Fellow of the Damon Runyon Cancer Research Foundation (DRG-2069-11 to J.P.S.-B.) and the National Science Foundation (W.A.P.).

Author information

Author notes

    • William A Pastor

    Present address: Department of Molecular, Cell and Developmental Biology, University of California at Los Angeles, Los Angeles, California, USA.

    • Edahí González-Avalos
    •  & Sini Rautio

    These authors contributed equally to this work.

Affiliations

  1. Department of Signaling and Gene Expression, La Jolla Institute for Allergy and Immunology, La Jolla, California, USA.

    • Ageliki Tsagaratou
    • , Edahí González-Avalos
    • , James P Scott-Browne
    • , Susan Togher
    • , William A Pastor
    •  & Anjana Rao
  2. Department of Computer Science, Aalto University School of Science, Aalto, Finland.

    • Sini Rautio
    •  & Harri Lähdesmäki
  3. Sanford Consortium for Regenerative Medicine, La Jolla, California, USA.

    • James P Scott-Browne
    •  & Anjana Rao
  4. Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, California, USA.

    • Ellen V Rothenberg
  5. Division of Pediatric Neurooncology, German Cancer Research Center (DKFZ), Heidelberg, Germany.

    • Lukas Chavez
  6. Department of Pharmacology and Moores Cancer Center, University of California at San Diego, La Jolla, California, USA.

    • Anjana Rao

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Contributions

A.R. and A.T. designed the study; A.T. performed all of the experiments, and A.R. and A.T. wrote the manuscript. E.G.-A. analyzed WGBS and ATAC-seq data sets under the supervision of L.C. and J.P.S.-B., respectively. S.R. analyzed RNA-seq and CMS-IP data sets under the supervision of H.L. S.T. helped with in vivo adoptive transfer experiments. W.A.P. generated the Tet3fl/fl mice. E.V.R. provided critical input and suggestions during the course of this study and helped write the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Anjana Rao.

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https://doi.org/10.1038/ni.3630

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