Figure 1.

AZA reduces proliferation but increases activation of T cells and increases Treg frequency in vitro. Fresh T cells from three healthy volunteers were incubated for 12 d in technical duplicates in the presence or not of 1 µM of AZA. (A) CFSE loaded T cells were incubated for 3 d in presence of AZA (bold) or PBS (dashed) and analyzed by flow cytometry (FCM). (B–D) FCM comparisons of KI67⁺ (B), CD25⁺HLA-DR⁺ (C) and CD25 MFI (D) of T cells on days +4, +8, +12 in presence or not of AZA (given from day 0 to day 8). (E) Evolution of CD4⁺CD25⁺FOXP3⁺ Treg cells frequency among cultured CD4⁺ T cells receiving AZA for 4 d (red), 8 d (green) or PBS for 8 d (blue) on days +4, +8, +12, analyzed by FCM (p-values report the comparison of each time point with day +4 from the corresponding condition). Data show median values of biologic triplicates and technical duplicates (n = 6) with interquartile range (*p < 0.05, **p < 0.005, ***p < 0.0005).