Functional assay of NS3 intracellular processing. (A) ELISPOT detection of IFN-γ release by NS3_1406-1415–specific CD8⁺ clone cocultured for 18 hours with DCs previously pulsed with NS3 protein for 16 hours and then washed and used in a cross-presentation assay. The results are expressed as mean ± SD of 10 independent experiments. (B-C) Evaluation of IFN-γ–forming spots in cocultures of IFN-DCs (B) or IL-4-DCs (C) incubated with the specific inhibitors before 16 hours of NS3 pulsing and then washed and used in a cross-presentation assay with NS3_1406-1415–specific CD8⁺ clone (mean ± SD values of 5 independent experiments). (D) IFN-γ analysis of DCs pulsed with NS3 protein for 1 hour and then washed and used after 3 days in a cross-presentation assay with NS3_1406-1415–specific CD8⁺ clone (mean ± SD; n = 4). Statistical analysis in all panels was performed by Mann–Whitney test (*P < .05).