Fig. 6

Specificity and safety of MSLN-CAR T cells. A The cytotoxicity of MSLN-CAR T cells on luciferase-expressing target cells was quantified after incubation for 20 h at an E:T ratio of 2:1. Three independent experiments were performed per condition. Statistics: two-tailed one-way ANOVA. Data are presented as the mean ± SD, n = 3; ***P < 0.001 vs CD19-CAR or NTD. B Quantification of retroviral vector copy number (VCN) in transduced T cells (CAR). Measured VCN per cell of MSLN-CAR T cells derived from three healthy donors above. Cells were assessed 96 h post transduction and the transduction MOI and efficiency were the same as the above. The experiment was conducted using three biologic replicates, each one including three technical replicates. C Levels of IFN-γ (left), and TNF-α (right) were measured by ELISA on serum collected from peripheral blood on three days after immune effector cell infusion. For statistical analysis of two groups an unpaired two sample t-test is used. Data are presented as the mean ± SD, n = 3. **P < 0.01 vs CD19-CAR. D Designated organs were analyzed after fluorescent staining of immune tissues on day 5 after intraperitoneal or intravenous administration of CAR T cells. Representative confocal microscopy images of CD3⁺ T cells in major organs (heart, liver, spleen, lung, kidney, brain) (n = 3 mice/group). Scale bar = 100 μm. The experiments were repeated twice, independently, with similar results