Fig. 5.

MICA immune complex formed with anti-MICA antibody clone 6E1 directly activates NK cells. a C1R-MICA*002 cell killing was assessed by co-culturing C1R-MICA*002 cell line with primary NK cells. The NK cells were pretreated with MICA-ECD alone, MICA-IC preformed with 6E1 (hIgG1, wildtype), MICA-IC preformed with 6E1 Fc effectorless mutant (hIgG1, N297G), or no treatment; NK cell killing of parental C1R cell line was used for comparison. Each data point represents the average of 2 technical replicates, the dataset is a representative of 3 independent experiments and p values were generated from unpaired t test. b NK cells were treated with MICA-ECD alone, MICA-IC preformed with 6E1 (hIgG1 wildtype) or MICA-IC preformed with 6E1 Fc effectorless mutant (hIgG1, N297G); IFN-γ and TNF-α secretion was analyzed using Luminex platform. Each data point represents average of 2 technical replicates, and the dataset is representative of 3 independent experiments and p values were generated from unpaired t test. c NK cells were cultured with MICA-IC preformed by 6E1 (hIgG1, N297G) that was bound to the goat anti-human Fc antibody coated to the surface of assay plate; IFN-γ and TNF-α secretion was analyzed using Luminex platform. Each data point represents average of 2 technical replicates (error bar represents SEM), and the dataset is a representative of 3 independent experiments. *p > = 0.05; **p < 0.05