Extended Data Fig. 10: Immunophenotype profiling of quad-CAR and quad-CAR triple-Switch T cells.
From: Leucine zipper-based immunomagnetic purification of CAR T cells displaying multiple receptors
a–n, Resting and BM185-CD20-stimulated quad-CAR and quad-CAR triple-Switch T cells (as depicted in Fig. 6), were analysed by multiparameter spectral flow cytometry. Cells were stimulated at 1:1 E:T ratio with BM185-CD20 for 24 h or left unstimulated without IL-2. a, t-Distributed Stochastic Neighbour Embedding (t-SNE) projection with flowSOM metaclusters depicting combined stimulated and unstimulated CD4+ quad-CAR and quad-CAR triple-Switch T cells (n = 3 concatenated technical replicates from triplicate wells). b, tSNE projections illustrating flowSOM metaclusters separated by stimulated and unstimulated quad-CAR and quad-CAR triple-Switch T cell populations. c, tSNE projections depicting endogenous PD-1 and PD-1-OX40 switch receptor expression. d, Population distributions of stimulated (top) and unstimulated (bottom) T cell products in metaclusters. e, tSNE projections of selected protein expression for combined stimulated and unstimulated quad-CAR and quad-CAR triple-Switch T cells. f, Heat map depicting Z-score-transformed median fluorescence intensity expression values depicted for proteins in stimulated and unstimulated quad-CAR and quad-CAR triple-Switch T cells. g, Histograms depicting protein expression for selected markers. h–n, Concurrent analysis of CD8+ T cell populations as in panels a–g. Data are representative of n = 2 donor experiments with triplicate wells. d,k, Mean of technical replicates and individual data replicates are depicted.
