K562 target cells were transduced with human CD19 and NucLight Red (Essen BioScience, Ann Arbor, MI, USA) and cocultured with various engineered T cells. Cytolytic activity was assessed by measuring the loss of viable target cells over a period of 3 days, as determined by red fluorescence signal (IncuCyte Live Cell Analysis System, Essen BioScience). The number of NLR+ target cells was determined every 2 hours. Normalized target cell numbers were generated by dividing target cell counts to cell counts at the start of each culture. To capture the cytolytic activity of CAR T cells against target cells over time, the area under the curve (AUC) was calculated. Where described, the “kill index” is the 1AUC value.

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