5-Azacytidine Promotes an Inhibitory T-Cell Phenotype and Impairs Immune Mediated Antileukemic Activity
Figure 3
CD8 T-cells show reduced cytotoxic function after 5-Aza treatment and upregulation of FOXP3. (a) CD3+ or CD8+ T-cells were isolated and treated with 5 μM or 20 μM or without 5-Aza for 48 h. Thereafter, T-cells were resuspended in fresh medium and cocultured with the target cell line HL60 for 4 h. LDH release was used as marker for cell death. Specific cytotoxicity was calculated as described by the manufacturer. Four independent experiments (all with an E : T ratio 10 : 1) are shown. (b) CD8+ T-cells were sorted and treated with 5-Aza (5 M and 20 M) for 48 h. mRNA levels of FOXP3 in CD8 cell compartment were assessed by qRT-PCR. (). (c) CD8+ T-cells were analyzed after treatment with or without 5-Aza, for their intracellular expression of FoxP3. A representative of three different experiments is shown.
