Extended Data Fig. 3: Manufacturing and in vitro validation of BTLA-deleted CART30 cells.
From: The BTLA–HVEM axis restricts CAR T cell efficacy in cancer
a, Schematic depicting timeline for CRISPR-edited human CAR T cell manufacturing. b, Prediction of off-targeting knockouts from optimized BTLA sgRNA, showing no additional targets. c, Representative analysis of indels in exon 2 of BTLA following CRISPR-Cas9 KO, using the TIDE (Tracking of Indels by DEcomposition) software (https://tide.nki.nl/). d, Expression of T1-159 CAR30 in primary human T cells at day 6 (de-bead) of expansion, measured via flow cytometry using an anti-G4S linker antibody. e, Expansion metrics of T1-159 CART30 for a single donor (ND518), including population doublings, total cells, cell size, and viability. f, Cytotoxicity of WT UTD, BTLA KO UTD, WT CART30, and BTLA KO CART30 against HDLM-2 at 48 h (n = 2 technical replicates per condition). g, Concentration of TNF in supernatant after 24 h of coculture with HDLM-2 (effector: target = 1, n = 3 technical replicates, representative of two independent biological donors). For all bar plots, data are presented as mean ± s.d. Statistical significance was determined by one-way ANOVA with post hoc Tukey tests (g). EP, electroporation.
