Dynamic CpG-DNA Methylation of Il10 and Il19 in CD4⁺ T Lymphocytes and Macrophages: Effects on Tissue-Specific Gene Expression

Abstract

The IL-10 family of cytokines consists of 9 members, including the immune-regulatory IL-10; Il19 is in close physical relationship with Il10 in the so-called IL-10 cytokine cluster on chromosome 1q32. While IL-10 is ubiquitously expressed, IL-19 expression is restricted to myeloid and epithelial cells. Little is known about molecular mechanisms that control tissue-specific expression of IL-10, and IL-19. Modifications in CpG-DNA methylation are a key mechanism in controlling transcription. Using bisulfite sequencing, we demonstrate that murine Il19 is methylated in CD4⁺ T lymphocytes. Macrophages display site-specific demethylation of Il19. The ubiquitously expressed Il10 gene is methylated to a lower degree and exhibits tissue-specific methylation patterns. DNA demethylation with 5-azacytidine resulted in an induction of IL-10, and IL-19 expression in CD4⁺ T cells, and CpG-DNA methylation through DNMT3a resulted in transcriptional silencing in macrophages. Thus, our findings suggest a role of CpG-DNA methylation in the regulation of Il10 and Il19.

Zusammenfassung

Die IL-10-Zytokinfamilie besteht aus 9 Mitglie­dern: dem immun-regulatorischen Zytokin IL-10; Il19 liegt gemeinsam mit Il10 im sog. IL-10- „Zytokincluster“ auf Chromosom 1q32. Während IL-10 von einer Vielzahl an Zellen exprimiert wird, ist die Expression von IL-19 auf myeloide und epitheliale Zellen beschränkt. Über die molekularen Mechanismen der gewebs­spezifischen Expression von IL-10 und IL-19 ist wenig bekannt. Modifikationen der CpG-DNA-Methylierung sind ein Schlüsselmechanismus der transkriptionellen Kontrolle. Durch Bisulfitsequenzierung zeigen wir hochgradige CpG-DNA-Methylierung des murinen Il19-Gens in CD4⁺-T-Lymphozyten und regionsspezifische Demethy­lierung in Makrophagen. Das ubiquitär expri­mierte IL-10- ist zu einem niedrigeren Grad me­thyliert. DNA-Demethylierung durch 5-Azacytidin resultiert in einer Induktion der IL-10- und IL-19-Expression in CD4⁺-T-Lymphozyten. CpG-DNA-Methylierung in Makrophagen durch DNMT3a resultiert in transkriptionellem Stillstand. Dies deutet auf eine Rolle der CpG-DNA-Methylierung bei der transkriptionellen Kontrolle von Il10 und Il19 hin.

Key words

IL-19 - IL-10 - DNA methylation - inflammation - gene regulation - chromatin

Schlüsselwörter

IL-19 - IL-10 - DNA Methylierung - Entzündung - Genregulation - Chromatin

• References

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