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Notch ligand Delta-like 4 induces epigenetic regulation of Treg cell differentiation and function in viral infection

Mucosal Immunology (2018) | Download Citation

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Abstract

Notch ligand Delta-like ligand 4 (DLL4) has been shown to regulate CD4 T-cell differentiation, including regulatory T cells (Treg). Epigenetic alterations, which include histone modifications, are critical in cell differentiation decisions. Recent genome-wide studies demonstrated that Treg have increased trimethylation on histone H3 at lysine 4 (H3K4me3) around the Treg master transcription factor, Foxp3 loci. Here we report that DLL4 dynamically increased H3K4 methylation around the Foxp3 locus that was dependent upon upregulated SET and MYDN domain containing protein 3 (SMYD3). DLL4 promoted Smyd3 through the canonical Notch pathway in iTreg differentiation. DLL4 inhibition during pulmonary respiratory syncytial virus (RSV) infection decreased Smyd3 expression and Foxp3 expression in Treg leading to increased Il17a. On the other hand, DLL4 supported Il10 expression in vitro and in vivo, which was also partially dependent upon SMYD3. Using genome-wide unbiased mRNA sequencing, novel sets of DLL4- and Smyd3-dependent differentially expressed genes were discovered, including lymphocyte-activation gene 3 (Lag3), a checkpoint inhibitor that has been identified for modulating Th cell activation. Together, our data demonstrate a novel mechanism of DLL4/Notch-induced Smyd3 epigenetic pathways that maintain regulatory CD4 T cells in viral infections.

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Acknowledgements

HT, DDN, MAS, and NWL designed the experiments. HT, AJR, DDN, and CM performed experiments. HT and NWL did data analysis and wrote the manuscript. We thank Dr. Matthew A Schaller and Consulting for Statistics, Computation, and Analytical Research (CSCAR) for consultations; Ivan Maillard for helpful discussions; Susan Morris, Lisa Riggs Johnson, for technical assistance; and Dr. Judith Connett for editing the manuscript. The manuscript was supported in part by NIH grant AI036302 (NWL).

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Author notes

    • Denise de Almeida Nagata

    Present address: Department of Cancer Immunology, Genentech, South San Francisco, CA, 94080, USA

    • Ivan P Maillard

    Present address: Department of Medicine, Abramson Family Cancer Research Institute, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA, 19104, USA

Affiliations

  1. Department of Pathology, University of Michigan, Ann Arbor, MI, 48109, USA

    • Hung-An Ting
    • , Denise de Almeida Nagata
    • , Andrew J Rasky
    • , Carrie-Anne Malinczak
    • , Matthew A Schaller
    •  & Nicholas W Lukacs
  2. Molecular and Cellular Pathology Program, University of Michigan, Ann Arbor, MI, 48109, USA

    • Hung-An Ting
    • , Carrie-Anne Malinczak
    •  & Nicholas W Lukacs
  3. Department of Internal Medicine, Life Sciences Institute, University of Michigan, Ann Arbor, MI, 48109, USA

    • Ivan P Maillard
  4. Department of Cell and Developmental Biology, Life Sciences Institute, University of Michigan, Ann Arbor, MI, 48109, USA

    • Ivan P Maillard

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The authors declare no competing financial interests.

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Correspondence to Nicholas W Lukacs.

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https://doi.org/10.1038/s41385-018-0052-1