Extended Data Fig. 5: Re-analysis of previous published flow cytometry data performed on patient 2 demonstrates the functional capacity of CD4⁺, CD8⁺, and double-negative CAR T-cells.

a, Gating strategy for the identification of CD4⁺, CD8⁺, and double-negative CAR T-cells. Flow cytometry data are from the functional experiment described by Porter et al. Sci Transl Med (2015), in which the authors stimulated cells from patient 2 with CD19 expressing K562 cells. Re-examination of the flow cytometry data identified prominent CD4⁺, CD8⁺, and double-negative CAR T-cell populations. Shown is representative gating at day 259, in which 31.2% of CAR T-cells were double-negative CAR T-cells. b, Line plots and box plots showing CAR-specific activation of CD4⁺, CD8⁺, and double-negative CAR T-cells supported by greater proportions of CAR⁺ T-cells expression MIP-1B and CD107a compared to CAR⁻ cells in response to CAR specific stimulation. Pairwise statistical testing was performed using the two-sided Welch’s t-test.