DCs incorporating CHP/HER2-147 effectively present HER2p63 peptide on HLA-A2402 molecules.

(A) HLA-A2402⁺ DC (1 × 10⁶) were cultured with 0.001 to 20 μg/mL CHP/HER2-147, CHP/CAB, HER2-147 protein, HER2p63, or HER2p780 at room temperature for 1 hour and were recultured for an additional hour at 37°C. DCs (1 × 10⁵cells/well) were incubated with 5 × 10⁴ cells/well Y.I.1a for 18 hours, and IFN-γ production was measured in duplicate using an ELISA kit. Data represent the average of duplicate measurements. (B) HLA-A2402⁺ (1 × 10⁶) and HLA-A2402⁻ DCs (1 × 10⁶) were cultured with 20 μg/mL protein CHP/HER2-147 or CHP/CAB for 3 hours at 37°C, washed, recultured in 10% FCS RPMI 1640 alone for an additional 18 hours, and incubated with the CTL clone, Y.I.1a, for 5 hours at an effector-target ratio of 2.5. Cytotoxic activity was measured using ⁵¹Cr release assay.