Extended Data Fig. 2: T cells from CR are enriched for serial killing, increasing mitochondrial and lysosomal size, and persistent migration.
(a) Schematic of a killing event at an E:T of 1:1 in which a CAR T-cell conjugates and kills a NALM-6 cell. The plot on the right shows the killing rate comparison between T cells from CR and PR/PD within all 1E:1T nanowells. Each dot (n) represents the frequency of killer T cells for each IP. Micrograph showing an example of 1E:1T killing event through the 6-hours (hh:mm) of time-lapse imaging from a CR IP. (b) Schematic of a killing event, showing the interaction parameters. tSeek defined as the time for CAR-T cell to find and conjugate to the NALM-6 cell. tconjugation is defined as the duration of CAR-T cell in stable conjugation with NALM-6 cell. tDeath is the time interval between the start of the conjugation and the apoptosis of the NALM-6 cell. Plots show the comparison between T cells from CR and PD/PR for these parameters. Each dot (n) represents the average value for all T cells within each IP. (c) Representative violin plot shows the interaction parameters from one responder IP: tSeek (n = 96 events), tconjugation (n = 94 events), tDeath (n = 30 events). The bar graph shows the killing frequencies of the same responder IP at an E:T of both 1:1 and 1:2. (d) Schematics and examples of serial killing, mono killing and no-killing events in nanowells with an E:T of 1:2. (e) Unsupervised hierarchical clustering based on parameters from TIMING, and confocal microscopy. Serial killing, migration, increasing mitochondrial and lysosomal volume were features associated with T cells from CR patients. (f) Cytotoxicity and motility correlations with CD4/CD8 ratio. Each point represents the average parameter for each IP (n = 9 patient IPs). Cytotoxicity is defined as the frequency of 1E:1T killing from TIMING. The Pearson’s correlation coefficient was calculated for CR and PR/PD IP.
