Effect of HDAC1 deficiency on in vitro Th2 and pathogenic Th2 differentiated murine CD4+ T cells
The differentiation of naïve CD4+ T cells into distinct effector T helper (Th) subsets is controlled by the chromatin state of the cells which in turn is regulated by various epigenetic mechanisms such as reversible lysine acetylation mediated by histone acetyltransferases and histone deacetylases (HDACs). In addition, Th2 cells can aquire pathogenic features, secreting pro-inflammatory factors and high levels of the cytokine IL-5. By using HDAC1 deficient naïve CD4+ T cells and establishing a protocol for differentiation of pathogenic Th2 cells, we could establish a critical role for HDAC1 in regulating the differentiation and pathogenicity of in vitro generated pathogenic Th2 cells Overall design: To study the function of HDAC1 in the differentiation of pathogenic Th2, naïve CD4+ T cells were isolated by magnetic cell sorting from CD4cre-HDAC1f/f (WT) and CD4cre+HDAC1f/f (KO) and differentiated into Th2 and pathogenic Th2 in vitro. Briefly, cells were activated with immobilized anti-CD3/CD28 in a polarizing cytokine milieu. Cells wer collected after 3 days in culture and RNA processed for RNA sequencing. We performed gene expression profiling by comparing WT Th2 to WT pathogenic Th2 conditions as well as KO Th2 to KO pathogenic Th2 and WT Th2.