Fig. 7
Immunoprofiling of engineered TRACCAR T cells activated by tumor cells in vitro. a Schematic showing the experimental design to investigate the effector and memory phenotype and cytokine secretion profile of activated engineered TRACCAR T cells. 106 TRACCAR T cells were mixed with a suspension of RAJI-luc tumor cells at E:T = 5:1 in a total volume of 40 mL of Xvivo-15 media supplemented with 5% AB serum and incubated in a GREX-10 device. The mixture was incubated for 24 h before adding 2 × 106 RAJI-Luc cells. The resulting cell mixture was incubated for 24 h and the same procedure was repeated to get a total of 4 RAJI cell challenges. The mixture was then incubated as is for 4 additional days and where spun down to recover the supernatant and determine the cytokine secretion profile of TRACCAR T cell. TRACCAR T cells were also analyzed by flow cytometry to determine the frequency of CD62L and CD45RA population within CD8+ CAR+ T cells (day 0 and day 5 post tumor cell-dependent activation). b Frequency of T cell subpopulation displaying CD62L+CD45RA+ (naive-like), CD62L+ CD45RA− (central memory), CD62L-CD45RA− (effector memory), CD62L-CD45RA+ (terminal effector) labeling and obtained at day 0 and day 5 post tumor cell-dependent activation for different engineered T cells. A linear mixed model, using lmer from the lme4 R package was used for statistical analysis. p-values are documented in Supplementary Table 1. c Cytokine profiling assay results obtained from supernatant of engineered T cell/RAJI cells cocultured for 5 days. The data shown in b and c represent the average of three experiments performed with cells engineered from three different donors. The mean of cytokine concentration fold change and the whiskers representing the maximum and minimum data points are illustrated in c. Two-way ANOVA was used in panel c for statistical analysis (p-value are indicated on the figures). Source data are provided as a Source Data file
