Fig. 1

MSLN-CAR T cells can efficiently lyse MUC16-positive human ovarian cancer cells. A Schematic illustration of a CD19-CAR and a MSLN-CAR T vector, respectively. B, C Quantitative analysis of MUC16 protein expression levels in ovarian cells (SKOV3, OVCAR3) by Western blotting (B) and flow cytometry (C). Tubulin was used as a loading control. Lane 1 = molecular marker; Lanes 2 = SKOV3 cell lysate; Lanes 3 = OVCAR3 cell lysate; Lanes 4 = MUC16 overexpression in 293T cell lysate. B and C consist of three technical replicates. The pictures represent one example of three technical replicates. D The cytotoxicity of MSLN-CAR T cells on ovarian cancer cell lines was quantified using a luciferase assay. Primary human T cells transduced with the indicated lentiviruses were co-incubated with target cells expressing luciferase at an effector to target (E:T) ratio of 2:1 for 20 h. Three independent experiments were performed. E Quantification of IFN-γ (left) and TNF-α (right) release in response to coculture with CD19-CAR or MSLN-CAR T cells at an E:T ratio of 2:1, as measured by ELISA. Data are presented as the mean ± SD, n = 3. F CAR T cells were co-incubated with target cells expressing luciferase at varying effector to target (E:T) ratios for 20 h. D–F consist of three biologic replicates and three technical replicates. Statistics: two-tailed one-way ANOVA. The results are presented as the mean volume ± SD; *P < 0.05, **P < 0.01, ***P < 0.001 vs CD19-CAR or NTD