Figure 5
#21-3BiTE shows potent T-cell dependent cytotoxicity against surviving-2B+ HLA-A*24+ tumor cells in vitro. (a) T2/A24 cells stably expressing luciferase were pulsed with the indicated peptide and then mixed with T-cells expanded from a HLA-A*24+ healthy donor at an E:T ratio of 10:1 in the presence of serial dilutions of #21-3BiTE. The reactions were allowed to proceed for 8 h. Target cell viability was determined by luciferase assay. Data are representative of two independent experiments. (b) HeLa/A24 or HeLa/A02 cells stably expressing luciferase were mixed with T-cells at E:T ratio of 10:1 in the presence of serial dilution of #21-3BiTE, and the reactions were allowed to proceed for 12 h. Relative cell viability values for HeLA cells is determined by luciferase assay. Data are representative of three independent experiments. (c) WiDr cells stably expressing luciferase were mixed with the T-cells at an E:T ratio of 10:1 in the presence of serial dilution of the indicated antibody, and the reactions were allowed to proceed for 14 h. Relative cell viability values for WiDr cells was determined by luciferase assay. Representative data from three independent experiments are shown. (d) Colo320DM cells were treated as in (c), and relative cell viability values was determined by luciferase assay. (e) WiDr cells mixed with the T-cells at an E:T ratio of 10:1 were incubated with 10 ng/mL of #21-3BiTE in the presence of serial dilutions of SV2B80-88/HLA-A*24 monomer, and the reactions were allowed to proceed for 14 h. Relative cell viability values for WiDr cells was determined by luciferase assay. Data are representative of two independent experiments. (f) Colo201 cells were treated as in (c), and the reactions were allowed to proceed for 14 h. Target cell viability was determined by luciferase assay. Relative cell viability values for target cells are represented as the average ± SD for three replicates/group (***p < 0.001).
