Suppression of cytokine production at 96 hours is predicable at 7 hours. (A) Absolute concentrations of IL-2 and IFN-γ in 96-hour supernatants of Tresps cultured alone or together with freshly isolated or GMP-Tregs at various ratios. (B) Suppression of IL-2 and IFN-γ in supernatants by Tregs. (C) Relationship between suppression of CD69 and CD154 expression at 7 hours and inhibition of IL-2 and IFN-γ at 96 hours. (A-C) Cumulative data from 4 independent experiments with each type of Treg showing freshly isolated Tregs (left panels) and GMP-Tregs (right panels). *P < .05, **P < .01, ***P < .001, ****P < .0001. †P < .05, ††P < .01, †††P < .001, and ††††P < .001 with respect to Tresp alone cultures (Tresp:Treg ratio of 1:0).