title: Epigenetic modification of the human CCR6 gene is associated with stable CCR6 expression in T cells.
abstract: CCR6 is a chemokine receptor expressed on Th17 cells and regulatory T cells that is induced by T-cell priming with certain cytokines, but how its expression and stability are regulated at the molecular level is largely unknown. Here, we identified and characterized a noncoding region of the human CCR6 locus that displayed unmethylated CpG motifs (differentially methylated region [DMR]) selectively in CCR6(+) lymphocytes. CCR6 expression on circulating CD4(+) T cells was stable on cytokine-induced proliferation but partially down-regulated on T-cell receptor stimulation. However, CCR6 down-regulation was mostly transient, and the DMR within the CCR6 locus remained demethylated. Notably, in vitro induction of CCR6 expression with cytokines in T-cell receptor-activated naive CD4(+) T cells was not associated with a demethylated DMR and resulted in unstable CCR6 expression. Conversely, treatment with the DNA methylation inhibitor 5'-azacytidine induced demethylation of the DMR and led to increased and stable CCR6 expression. Finally, when cloned into a reporter gene plasmid, the DMR displayed transcriptional activity in memory T cells that was suppressed by DNA methylation. In summary, we have identified a noncoding region of the human CCR6 gene with methylation-sensitive transcriptional activity in CCR6(+) T cells that controls stable CCR6 expression via epigenetic mechanisms.
authors.affiliation: Charité University Medicine Berlin, Research Centre for ImmunoSciences, Germany.
authors.forename: Svenja
authors.initials: S
authors.lastname: Steinfelder
authors.forename: Stefan
authors.initials: S
authors.lastname: Floess
authors.forename: Dirk
authors.initials: D
authors.lastname: Engelbert
authors.forename: Barbara
authors.initials: B
authors.lastname: Haeringer
authors.forename: Udo
authors.initials: U
authors.lastname: Baron
authors.forename: Laura
authors.initials: L
authors.lastname: Rivino
authors.forename: Bodo
authors.initials: B
authors.lastname: Steckel
authors.forename: Andreas
authors.initials: A
authors.lastname: Gruetzkau
authors.forename: Sven
authors.initials: S
authors.lastname: Olek
authors.forename: Jens
authors.initials: J
authors.lastname: Geginat
authors.forename: Jochen
authors.initials: J
authors.lastname: Huehn
authors.forename: Alf
authors.initials: A
authors.lastname: Hamann
mesh_terms: D002469:Cell Separation; D019175:DNA Methylation / Q000235:genetics*; D044127:Epigenesis, Genetic / Q000235:genetics*; D005434:Flow Cytometry; D015870:Gene Expression; D005786:Gene Expression Regulation / Q000235:genetics*; D006801:Humans; D016133:Polymerase Chain Reaction; D054399:Receptors, CCR6 / Q000235:genetics*; D013601:T-Lymphocytes / Q000378:metabolism*; D014162:Transfection
chemical_list: C106311:CCR6 protein, human; D054399:Receptors, CCR6