Cross-presentation of NY-ESO-1 formulations by MoDCs. (A) MoDCs of an HLA-A2⁺ patient with melanoma were pulsed with 10 μg/mL of either NY-ESO-1 protein, NY-ESO-1/IC (ES121), NY-ESO-1/IMX, or IMX alone and cultured overnight before coculture with autologous NY-ESO-1_157-165–specific CD8⁺ T cells. IFN-γ–producing T cells were quantified by ELISpot assay. Peptide-pulsed MoDCs served as a positive control. (B) MoDCs from an HLA-A2⁺ healthy donor were pulsed with NY-ESO-1 formulations as described for panel A. Induction of IFN-γ production by NY-ESO-1–specific CD8⁺ T cells was quantified by intracellular cytokine staining (ICS). Data are representative of 6 experiments. (C) Comparison of 3 different NY-ESO-1/IC formulations generated by incubating NY-ESO-1 protein with anti–NY-ESO-1 mAbs, ES121 or E978, or with serum from a patient with high serum titers of anti–NY-ESO-1 antibodies. Induction of IFN-γ production by NY-ESO-1–specific CD8⁺ T cells was quantified by ICS. Data are representative of 3 experiments.