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. 2012 Feb 28;16(3):569–581. doi: 10.1111/j.1582-4934.2011.01343.x

Fig 3.

Fig 3

NK-92-scFv(ch14.18)-ζ cells display enhanced cell killing activity towards established GD2-expressing NB cells. (A) Cytotoxic activity of NK-92-scFv(ch14.18)HL-ζ cells towards UKF-NB3, Kelly and SK-N-SH NB cells was determined in FACS-based cytotoxicity assays at different effector to target ratios (E/T). Parental NK-92 cells were included for comparison. To investigate dependence of cell killing on GD2 recognition, GD2 on Kelly NB cells was blocked with parental murine anti-GD2 antibody 14G2a prior to co-culture with NK-92-scFv(ch14.18)HL-ζ as indicated. Data are represented as mean ± S.D. (B) Cytotoxic activity of NK-92-scFv(ch14.18)HL-ζ towards LAN-1 NB cells was determined in 51Cr-release assays at an E/T ratio of 6.3:1. Parental NK-92 cells were included for comparison (left panel). To block interaction of CAR with GD2 on the target cell surface, LAN-1 and NK-92-scFv(ch14.18)HL-ζ cells were co-cultured in the presence of 10 μg/ml of anti-idiotype antibody 1A7 (right panel). Data are represented as mean ± S.D. (C) Cytotoxic activity of NK-92-scFv(ch14.18)HL-ζ and NK-92-scFv(ch14.18)LH-ζ towards BE(2)C NB cells was investigated by microscopical analysis after 18 hrs of co-culture at low E/T ratios of 1:1 and 0.3:1. Parental NK-92 cells were included for comparison. Control cells were incubated in the absence of NK cells, or were treated with 8 μM of the apoptosis-inducing drug staurosporine as indicated. Representative fields are shown.