Extended Data Fig. 1: In vitro and in vivo characterization of 19-28ζ and 19-BBζ CAR T cells.
From: CAR T cell trogocytosis and cooperative killing regulate tumour antigen escape
a, Schematic representation of the CD19 CARs. EC, extracellular domain; scFv, single chain variable region specific for anti-CD19; TM, transmembrane domain. b, c, Percentage of CAR+ T cells (left) and MFI of T cell surface CAR expression (right). b, CAR expression analysis performed 4 days after T cell transduction (n = 9 donors). c, CAR expression analysis performed 7 days after stimulation on NIH/3T3 cells expressing CD19 (n = 10 donors). d, Luciferase-based 18-h cytotoxic assay using NALM6 target cells at several E:T ratios (n = 3 donors). e, Kaplan–Meier survival analysis of NALM6 cell-bearing mice treated with 1 × 106, 0.4 × 106 or 0.2 × 106 transduced CAR T cells. Data are pooled from two independent experiments for the 0.2 × 106 CAR T cell dose. Corresponding bioluminescence imaging data are presented in Fig. 1a. f, Representative FACS profile of NALM6 and CAR T cells. NALM6 cells are GFP+; CAR T cells are LNGFR+. Absolute counts of NALM6 cells (left) and CAR T cells (right) at day 14 after infusion (n = 4 mice per group). g, Left, EOMES/T-bet ratio of CAR T cells. Right, fraction of CAR T cells expressing PD-1, LAG-3 and TIM-3 at day 14 after T cell infusion (n = 4 mice per group). In f and g, cells were isolated from the bone marrow of mice treated with 0.2 × 106 CAR T cells. h, Representative FACS profile of cells obtained by the time of relapse, 39 days after T cell infusion (19-28ζ, representative of 3 mice; 19-BBζ, representative of 9 mice). NA, not available (no detectable cells). P values were determined by two-sided Mann–Whitney U-test (b, c, f and g), or log-rank Mantel–Cox test (e). Data are mean ± s.e.m.
