Methylation of FOXP3 in regulatory T cells is related to the severity of coronary artery disease

Abstract

Objectives: Regulatory T (Treg) cells have been shown to play a protective role in experimental atherosclerosis. However, it is unclear whether Tregs can protect from rupture of vulnerable plaque in patients with atherosclerosis. Demethylation of the DNA encoding the transcription factor forkhead box P3 (FOXP3) was found to be essential for the stable maintenance of the suppressive properties of Tregs. We aimed to evaluate Treg levels in patients with acute coronary syndrome (ACS) using a method based on Treg-specific DNA demethylation within the FOXP3 gene.

Methods and results: Peripheral blood was collected to determine Treg levels by PCR-based DNA methylation analysis. We found that Treg levels were decreased in patients with ACS compared with normal coronary controls. The decrease in Tregs was associated with the severity of the ACS. Furthermore, up-regulation of DNA-methyltransferases was detected in CD4(+)CD25(+) Tregs obtained from ACS patients as compared to those from normal coronary controls. A dose-dependent increase in the methylation of the Treg-specific demethylated region in FOXP3 was observed in cultures of PBMCs with ox-LDL. Moreover, the ox-LDL-induced Treg effects could be restored by loading (-)-epigallocatechin-3-gallate, a methyltransferase inhibitor. Treatment of CD4(+)CD25(+) Tregs with ox-LDL resulted in a 41% increase in the methylation of FOXP3, a 66% of reduction in FOXP3 mRNA expression, and an increase in the expression of DNA methyltransferase 3a as well as 3b.

Conclusions: Our data demonstrate that reduction in Treg cells is associated with ACS in atherosclerotic patients. Epigenetic suppression of FOXP3 might lead to down-regulation of Treg cells, and in turn increase the risk of ACS.