Abstract
Neutrophil extracellular traps (NETs; NETosis) is known as one of innate immunity system in neutrophil. However, these have been few reports about the relationship between epigenetics and NETs induction. In this study, we elucidate the mechanism of NETosis in terms of epigenetics, especially DNA methylation, on neutrophil differentiation. HL-60 cells are differentiated into neutrophil-like cells (nHL-60) by the treatment with 1.25% DMSO for 72 hours, and we confirmed by staining DNA used SYTOX green that NETosis is induced by the treatment with A23187, calcium ionophore. nHL-60 treated with 5-azacytidine (Aza), DNA methyltransferase 1 (DNMT1) inhibitor, increased A23187-induced NETosis. Interestingly, nHL-60 treated with Aza induced NETosis without the treatment with A23187. The Aza-treated nHL-60 increased peptidylarginine deiminase 4 (PAD4), which converted arginine to citrulline, expression, and citrullinated H3 did not increase by Aza. Moreover, NETosis is known to be involved in not only PAD4 but also myeloperoxidase (MPO) and reactive oxygen species (ROS), however, the treatment with Aza did not increase MPO expression and ROS induction. Therefore, these reports suggest that DNA demethylation by DNMT1 inactivation in neutrophil induce ROS-independent NETosis.
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