Extended Data Fig. 7: Characterization of expanded NK cells from Opt-Cs and Sub-Cs.
Clinical CAR19/IL-15 NK cells from Opt-Cs and Sub-Cs were used in panels a-c. Expanded NT-NK cells from an independent cohort of CBUs were used in panels d-f. (a) Bar graph showing the percentage of CAR expression in clinical CAR19/IL-15 NK cells from Sub-Cs (n = 21 donors) or Opt-Cs (n = 16 donors). (b) Cumulative population doublings (PDs) for CAR-NK cells from Sub-Cs (n = 21 donors) or Opt-Cs (n = 16 donors) expanded with K562-based feeder cells and IL-2 (200 U/mL). (c) SPADE analysis of CyTOF data showing the phenotype of the CAR19/IL-15 NK cells (expanded for 14–21 days) from Sub-Cs (n = 17 donors) or Opt-Cs (n = 8 donors). Frequencies of each cluster (1–6) are indicated; size and color of nodes represent numbers of clustered cells. (d) Cumulative PDs of NT-NK cells derived from an independent cohort of Sub-Cs vs. Opt-Cs (n = 4 donors per group). (e) Tumor rechallenge assay where NT-NK cells were rechallenged with RajimCherry at 5:1(E:T ratio). Tumor cells (16,700 cells) were added every 2 days for 8 days and tumor cell killing was measured by the tumor cell index. The bar plots show the AUC of tumor cell index (n = 3 donors per group). (f) Representative images from the serial tumor rechallenge assay from the experiment described in panel e. (g) Bar graph showing the PSI of NT-NK cells secreting different cytokines after Fc-CD16 stimulation (n = 3 per group). (h) OCR as a surrogate for oxidative phosphorylation (OXPHOS) was performed on NT-NK cells that were expanded from a subset of Sub-Cs and Opt-Cs (n = 3 donors each). The results of the mito stress test (left), and the bar graphs of basal respiration (middle) and maximal respiration (right) are presented; Oligo: Oligomycin, Rot/AA: Rotenone/Antimycin A. P-values were determined by two-tailed Student’s t test in panels a,e,h, or two-tailed one-way ANOVA in panels b,d,g. Each symbol represents an individual donor, data are shown as mean + s.e.m.
