De Novo DNA Methylation Is Required to Restrict T Helper Lineage Plasticity

Abstract

Naïve CD4+ T cells are highly plastic and can differentiate into discrete lineages with unique functions during an immune response. Once differentiated, helper T cells maintain a stable transcriptional memory of their initial lineage choice and resist redifferentiation. During embryogenesis, de novo DNA methylation operates on the hypomethylated genome of the blastocyst to achieve tissue-specific patterns of gene expression. Similarly, the ifn γ promoter is hypomethylated in naïve T cells, but Th2, Th17, and iTreg differentiation is accompanied by substantial de novo DNA methylation at this locus. To determine whether de novo DNA methylation is required to restrict T helper lineage plasticity, we used mice with T cell-specific deletion of the methyltransferase DNMT3a. Induction of lineage-specific cytokines occurred normally in the absence of DNMT3a, however, DNMT3a-deficient Th2, Th17, and iTreg completely failed to methylate the ifn γ promoter. This was accompanied by an increase in the transcriptionally permissive trimethyl H3K4 mark, and a reduction in inhibitory H3K27 methylation at the ifn γ locus. Failed de novo methylation resulted in failed silencing of the ifn γ gene, as DNMT3a-deficient Th2, Th17, and iTreg cells produced significant levels of IFNγ following restimulation in the presence of IL-12. Therefore, DNMT3a-mediated DNA methylation restricts T helper plasticity by establishing an epigenetically silent chromatin structure at regulatory regions of the ifn γ gene. Background: T cells undergo lineage commitment during an immune response and "remember" their lineage choice after the instructive signals cease. Results: T cells lacking a de novo DNA methyltransferase fail to silence the ifn γ gene. Conclusion: DNMT3a opposes T cell trans -differentiation by epigenetically silencing "off-lineage" genes. Significance: The proper control of inflammatory cytokine gene expression is crucial for immune homeostasis.