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. Author manuscript; available in PMC: 2019 Jul 1.
Published in final edited form as: Cancer Immunol Res. 2018 May 16;6(7):776–787. doi: 10.1158/2326-6066.CIR-17-0649

Figure 6. Assessment of cytotoxicity and IFNγ production in the presence of the CS1-NGK2D biAb.

Figure 6

(A) MM.1S cell line was cocultured with IL2-primed PBMCs, T cell- depleted PBMCs, or NK cell-depleted PBMCs at an E:T ratio of 10:1 for 24 hours in the presence of biAb. All depletions were from negative depletions. Specific lysis induced from the control biAb and CS1-NKG2D biAb on each effector was compared as shown in dashed (PBMCs), green (T cells), and red (NK cells). (B) IL2-primed PBMCs, purified T cells, and purified NK cells were cocultured with the MM.1S MM cell line for 24 hours in the presence of biAb. CD3+ T cells and CD56+CD3− NK cells were sorted. E:T ratio for NK cells: 0.5:1; E:T ratio for PBMCs and T cells: 10:1. (C) Effect of a NKG2D blocking antibody on the enhanced cytotoxicity of IL2-primed PBMCs induced by the CS1-NKG2D biAb against the MM.1S MM cell line at an E:T ratio of 10:1 for 24 hours. The effectors were pretreated with the blockade before coculturing the targets. (D) IFNγ production of the PBMCs and NKG2D-depleted PBMCs against the MM.1S MM cell line in the presence of biAbs. The E:T ratio is 1:1. (E) Cytotoxicity assay of the IL2-primed PBMCs against CS1− MOLT-3 cell line and CS1 overexpressed MOLT-3 cell line. The dose of biAbs for all experiments was 50 μg/mL. Results show specific lysis mean±S.E.M. of five independent experiments. NS: not significant; *p<0.05; **p<0.01; ***p<0.001 using Student’s t test for two-group comparison and linear mixed model for multiple group comparison.