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. 2017 Feb 24;8:175. doi: 10.3389/fimmu.2017.00175

Figure 2.

Figure 2

A dual targeting triplebody ULBP2-aCD19-aCD33 mediates NK cell-dependent killing of antigen loss variants. (A) NK cells were purified from healthy donor by negative selection and were primed by IL2 (200 U/ml) + IL15 (10 ng/ml) cytokines for 15–18 h (overnight). Next day, primed NK cells were incubated with DiR dye-labeled BV173 cells at indicated effector to target (E:T) ratio for 3 h. The incubation was continued either alone (No construct) or in the presence of 100 nM of immunoligand (U-19: ULBP2-aCD19, U-33: ULBP2-aCD33, U-19-33: ULBP2-aCD19-aCD33). After incubation, 7-AAD was added and 7-AAD-positive cells within DiR-positive gate indicated dead BV173 cells. One representative toxicity assay is shown. (B) Cumulative analysis of four independent toxicity assays at 2.5:1 (E:T) ratio (N = 4; each N represents an independent healthy NK cell donor). Error bars indicate SEM and statistical analysis by one-way ANOVA.