chNKG2D- or chDap10-modified T cells produce large amounts of Th1 cytokines and proinflammatory chemokines after coincubation with NKG2D ligand–expressing tumor cells. The NKG2D ligand expression on various target cells is shown in panel A. Cells were stained with mouse NKG2D Ig (open curve) or control Ig (shaded curve). Bar labeled M1 designates positive events. For detection of IFN-γ (B), T cells (105) were cocultured with 105 target tumor cells RMA (▦), RMA/Rae-1β (▪), RMA/H60 (▨), and YAC-1 (▤) or media alone (□) for 24 hours. Concentrations of IFN-γ in supernatants were determined by ELISA. For detection of other cytokines (C) and chemokines (D), irradiated (100 Gy) tumor cells instead were mixed with T cells for 3 days. Bio-plex assays were performed to measure the levels of GM-CSF (▧), IL-3 (▩), IL-5 (▦), IL-10 (▪) (shown in C), CCL3 (▪), and CCL5 (□) (D).
