To assess the killing capacity of T cells, 5000 51Cr-labeled target cells were cocultured with 50,000, 25,000, 12,500, or 6250 effector cells in 200 μl of T cell medium + IL-2 (100 IU/ml). Cells were incubated in 96-well V-bottomed plates (Greiner Bio-one, Kremsmünster, Austria) for 4 hours. Fifty microliters of supernatant was transferred into Isoplate-96 HB plates (PerkinElmer, Massachusetts, USA). After addition of 150 μl of scintillation cocktail (OptiPhase SuperMix, PerkinElmer), samples were incubated overnight at room temperature. Counts were acquired using a 1450 MicroBeta TriLux scintillation counter (PerkinElmer). Percentage killing was calculated using the following formula:%killing=(Condition Cr51 releasebackgroundMaximal Cr51 releasebackground)*100

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