5-Azacytidine Promotes an Inhibitory T-Cell Phenotype and Impairs Immune Mediated Antileukemic Activity
Figure 1
5-Azacytidine reduces T-cell proliferation mainly by inhibition of CD8 T-cell proliferation by p15 upregulation. (a) T-cells were isolated from buffy coats and cultured for one week in presence of IL-2. 12 h before 5-Aza treatment cells were seeded in fresh medium. T-cells were treated with 5 M or 20 M 5-Aza or left untreated as control. Numbers of viable cells were determined every 12 h, over a period of 72 h by Trypan blue exclusion. Depicted results show mean value of triplicates. (, ). (b) CD3, CD4, and CD8 T-cells were sorted and treated with (5 μM or 20 μM) or without 5-Aza. p15 mRNA levels of the different subsets were analyzed by qRT-PCR. Data show mean values of triplicates. (, , not significant (n.s.)). (c) After 5-Aza for 48 h T-cell subsets were analyzed by FACS with mAB against CD3, CD4, and CD8. CD4/CD8 ratios were indicated for each treatment group. Data represent mean value of four independent experiments with SD.
