5-Azacytidine Promotes an Inhibitory T-Cell Phenotype and Impairs Immune Mediated Antileukemic Activity
Figure 2
Treatment with 5-Aza induces FOXP3+ Treg and upregulates immunomodulatory cytokines. (a) CD3+ and CD4+ T-cells were isolated and cultured for one week in the presence of IL2. Thereafter, cells were treated with the indicated dosages of 5-Aza. After 48 h mRNA was isolated and FOXP3 expression levels were analyzed in the CD3+ and CD4+ T-cell subset. Data show mean value with SD. (, ). (b) T-cells were analyzed by FACS after the described treatment with or without 5-Aza for the expression of FoxP3. CD4+/CD25+/FoxP3+, triple positive cells were considered as Tregs. A representative example of three independent experiments is shown. (c) The marker combination CD4+/CD25hi/CD127lo was used to confirm the results of (b). Data show mean value with SD of three different experiments. (d) mRNA of 5-Aza treated or control CD3+ T-cells were isolated and mRNA levels of IL10 and TGF-β were measured. Data shown as fold pattern gene induction. (, ).
