CTL lines generated from HLA-A2+ normal donors after in vitro stimulation with autologous DCs pulsed with PCI-13 lysates mediate the HLA-A2-restricted killing of HNC targets. Normal HLA-A2+ donor PBMCs were stimulated twice with autologous DCs preloaded with lysate derived from the PCI-13 cell line and subsequently with irradiated PCI-13 cells on a weekly regimen as outlined in the Materials and Methods. On day 32-37 of culture, CD8+ T cells were enriched and analyzed for their ability to lyse PCI-13 target cells in the absence or presence of blocking anti-HLA-A2 mAb MA2.1 in 4 h cytotoxicity assays at the indicated E/T ratios. Targets included the HLA-A2+ HNC cell lines PCI-13 and PCI-1, and the HLA-A2-negative HNC cell line SCC-74. Data are from one representative assay of 2 performed.