Epigenetic regulation of IL-12-dependent T cell proliferation

Matthew Schaller

Toshihiro Ito

Ronald Allen

Danielle Kroetz

Nicolai Kittan

Catherine Ptaschinski

Karen Cavassani

William Carson

Nuria Godessart+4 authors

Supplementary Material

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*Department of Pathology, University of Michigan Medical School, Ann Arbor, Michigan, USA; ^{Department of Immunology, Nara Medical University, Nara, Japan; and}^{Dermatology Research, Almirall, S.A., St Feliu de Llobregat, Spain}

^{Correspondence: University of Michigan, 109 Zina Pitcher Pl., Ann Arbor, MI 41809, USA. E-mail:}ude.hcimu@ellahcsm

Received 2014 Aug 5; Revised 2015 May 15; Accepted 2015 May 18.

Abstract

It is well established that the cytokine IL-12 and the transcription factor STAT4, an essential part of the IL-12 signaling pathway, are critical components of the Th1 differentiation process in T cells. In response to pathogenic stimuli, this process causes T cells to proliferate rapidly and secrete high amounts of the cytokine IFN-γ, leading to the Th1 proinflammatory phenotype. However, there are still unknown components of this differentiation pathway. We here demonstrated that the expression of the histone methyltransferase Mll1 is driven by IL-12 signaling through STAT4 in humans and mice and is critical for the proper differentiation of a naïve T cell to a Th1 cell. Once MLL1 is up-regulated by IL-12, it regulates the proliferation of Th1 cells. As evidence of this, we show that Th1 cells from Mll1^+/− mice are unable to proliferate rapidly in a Th1 environment in vitro and in vivo. Additionally, upon restimulation with cognate antigen Mll1^{, T cells do not convert to a Th1 phenotype, as characterized by IFN-γ output. Furthermore, we observed a reduction in IFN-γ production and proliferation in human peripheral blood stimulated with tetanus toxoid by use of a specific inhibitor of the MLL1/menin complex. Together, our results demonstrate that the}MLL1 gene plays a previously unrecognized but essential role in Th1 cell biology and furthermore, describes a novel pathway through which Mll1 expression is regulated.

Keywords: trithorax, Th1 differentiation, histone methylation, STAT4

Abstract

F, forward; Hs, Homo sapiens; Mm, Mus musculus, R, reverse.

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ACKNOWLEDGMENTS

Funding for this research was provided by U.S. National Institutes of Health National Heart, Lung, and Blood Institute Grants HL112897, HL31237, and HL89216. The authors thank the Pathology Flow Cytometry Core for its help and support, Lisa Johnson for preparation of histological samples, and Dr. Judith Connett for her critical reading of the manuscript.

ACKNOWLEDGMENTS

Glossary

^−/−	deficient
BCG	Bacillus Calmette-Guerin
BMDC	bone marrow-derived dendritic cell
CCND	cyclin D
CD62L	cluster of differentiation 62 ligand
Cdkn	cyclin-dependent kinase inhibitor
ChIP	chromatin immunoprecipitation
DC	dendritic cell
EDU	5-ethynyl-2′-deoxyuridine
H3K4Me3	histone H3 lysine 4 methylation
IRF	IFN regulatory factor
MHC II	MHC class II
MLL1	mixed lineage leukemia 1
PPD	purified protein derivative
qPCR	quantitative PCR
rh	recombinant human
Tbx21	T-box 21
WT	wild-type

Glossary

Footnotes

The online version of this paper, found at www.jleukbio.org, includes supplemental information.

Footnotes

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