Ad.DCs induce NK cell activation. NK cells and DCs were cultured alone or cocultured together at 1:1 ratio for 24 hours and subsequently examined. (A-B) iDCs, iDCs transduced with 3 different viral loads (100, 500, and 1000 MOI: Ad.DCs 100, Ad.DCs 500, Ad.DCs 1000, respectively) and LPS/IFN-γ-mDCs were assessed. (C-E) iDCs, mDCs, and Ad.DCs transduced with 500 MOI of AdV were examined. (A) CD69 expression on CD56^dimCD16⁺ and CD56^highD16⁻ NK^high cell subsets was measured by multicolor FACS analysis, and (B) secreted IFN-γ was tested in cell culture supernatants by multiplex cytokine assays. (C) NK cells from DC/NK cell cocultures were tested for proliferation by CFSE dilution assay using FACS analysis. (D-E) NK cells from DC/NK cell cocultures were tested for the ability to kill K562 and DAUDI tumor cell targets by CTO-based cytotoxicity assay, respectively. (A) Data are mean fluorescence intensities (MFI) for CD69. Bars represent the coefficient of variance for 5000 events acquired. (B) Data are mean ± SEM IFN-γ (pg/mL) of duplicate tests. (D-E) Data are LU₂₀/10⁷ NK cells based on the percentage of CTO⁺7-AAD⁺ cells. (A-B) Data are representative of 10 experiments. (C-E) Data are representative of 3 experiments.