In vivo DC maturation in response to α-GalCer–loaded allogeneic fibroblasts. (A) Several DC maturation markers were evaluated 12 hours after intravenous injection of NIH3T3 or CD1d^hi-NIH3T3 cells with or without α-GalCer. (B) Expression of CD70 on DCs was analyzed 12 hours and 40 hours after immunization. (C) IL-12 production from CD11c⁺ DCs in vivo was evaluated by intracellular staining 4 hours after giving mice CD1d^hi-NIH3T3/Gal or NIH3T3 cells as previously reported.^43,54  (D) To determine whether the T-cell response seen in mice immunized with CD1d^hi-NIH3T3/Gal-ova is dependent on DCs, 2 × 10⁶ CFSE-labeled OT-I T cells were transferred before immunization in wild-type (WT) mice or diphtheria toxin (DT)–treated CD11c-DTR mice (Document S1). OT-I proliferation was evaluated by dilution of CFSE-labeled cells 3 days later. Data are representative of 2 independent experiments with 2 mice in each group.