Fig. 1 | Signal Transduction and Targeted Therapy

Fig. 1

From: IL-24 armored CAR19-T cells show enhanced antitumor activity and persistence

Fig. 1

Enhanced antitumor activity and persistence of CAR19-IL-24-T cells. a Schematic diagram of FMC63scFv-28ζ (CAR19) and CAR19-IL-24. SP, signal peptide; 2A, 2A “self-cleaving” peptide; EGFP, enhanced green fluorescent protein. b Representative flow cytometry data showed co-expression of CAR19 and EGFP in CAR19-T cells and CAR19-IL-24-T cells at days 10 post-transduction. CAR-T cells were confirmed by GFP and labeling with Alexa Fluor 647-AffiniPure F(ab’)2 Fragment Goat Anti-Mouse IgG (H+L) that recognizes scFv portion of the CAR19. c The cytotoxicity of T cells to target cells was measured with CytoTox 96® Non-Radioactive Cytotoxicity Assay. T cells were cocultured with K562 cells (CD19−) or K562-CD19 cells (CD19+) for 18 h at the indicated ratios. d Quantification and statistical analysis of residue Nalm6 cells, Raji cells, and K562-CD19 cells ratio in living cells subset during coculture. e Changes in GFP+ ratio of CAR19-T cells and CAR19-IL-24-T cells after cocultured with tumor Nalm6 cells, Raji cells and K562-CD19 cells at ratio = 1:1, respectively. f Quantification and statistical analysis of the apoptosis of GFP+ T cells in CAR19-T group and CAR19-IL-24-T group after challenged with Nalm6 cells, Raji cells, and K562-CD19 cells, respectively. g–j T cells were cocultured with tumor cells at ratio 1:1 for 24 h, without exogenous rIL-2. The supernatant was collected for ELISA assay of IL-24 (g), IL-2 (h), TNF-α (i) and IFN-γ (j) according to the manufacturer’s suggestions. k Analysis of Burkitt lymphoma tumor volumes. NSG mice were inoculated with Raji tumor cells (106 cells/mouse) at day 0, then treated with NT cells, CAR19-T cells and CAR19-IL-24-T cells (normalized to 5% CAR19+, 3 × 107 cells/mouse, n = 3/group) at day 8, respectively. All data with error bars were analyzed with GraphPad Prism8 and presented as mean ± SEM. e, f Unpaired and non-parametric Mann–Whitney test with two-tailed, (c, d, gk) One-way analysis of variance with Bonferroni correction. ns not significant; p > 0.05; *p < 0.05, **p < 0.01, ***p < 0.001

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