L1 Cell Adhesion Molecule-Specific Chimeric Antigen Receptor-Redirected Human T Cells Exhibit Specific and Efficient Antitumor Activity against Human Ovarian Cancer in Mice
Fig 5
CE7R+ T Cells Specifically Recognized and Killed L1-CAM-Expressing Primary Ovarian Cancer Cells Derived from Malignant Ascites.
A, Representative primary culture of ovarian cancer cells derived from patient ascites depicting typical epithelial cobblestone morphology. B, Representative cytogenetic analysis of primary culture of ovarian cancer cells derived from patient ascites. Stemline: 37, X, -X, der(2)t(2;6)(q35;p11), -4, del(5)(q13q33), -6, del(8)(p21p23), 9, del(10) (q24q26), der(11)t(11;?15) (p15;q15), der(12)t(12;?13) (q24.3;q22), -13, -16, -17, -18, 19, der(22)t(12;22)(q11;p13). C, Representative images of fluorescence in situ hybridization for cultured primary ovarian cancer cells in comparison to original ascites cells using probes for 13q14.3 (red) and 13q34 (green), 5p15 (red) and 5q31 (green), or P16 (red) and ABL (green). D, Flow cytometric examination of cell-surface L1-CAM expression on the primary cultured, ascites-derived cancer cells compared to the BE2 neuroblastoma, OVCAR-3 ovarian cancer, and D283 medulloblastoma cell lines. Mean fluorescent intensity (MFI) and percentages of cells with positive staining (%) over secondary reagent alone are indicated. E, CE7R+ T cells were co-cultured overnight with the indicated tumor lines at a 10:1 ratio and supernatants were analyzed for IFN-γ and TNF-α levels by cytometric bead array. Means + S.E.M. of triplicate wells are depicted. F, CE7R+ T cells against the indicated cancer cell lines targets was determined by 4-hr 51Cr-release assay. Mean % chromium release ± SD of triplicate wells are depicted.