Human CD8 T cell IL2 deprivation time course

IL2 deprivation time course for day 8 human CD8 T cells activated for 3 days with TransAct T-cell activation beads. PBMC (Cambridge Bio) were defrosted and CD8+ T-cells isolated using a human CD8+ T-cell isolation kit. CD8+ T-cells were activated at 1 x 106 cells/mL with 1 in 100 TransAct T-cell activation beads (Miltenyi) and 20 ng/mL IL-2 in complete RMPI (RMPI 1640, 10% FBS, penicillin/streptomycin) for three days. Cells were then washed out of activation and then maintained in complete RMPI with 20 ng/mL IL-2 for five days. To remove IL-2 for IL-2 deprivation, cells were washed twice in pre-warmed media, then replated in complete media without IL-2. Cells were deprived of IL-2 for 1hr, 3hrs, 5hrs, 8hrs, 24hrs or 48hrs.