Specific lysis of the LP-1 cell line by CTLs induced by DCs transfected with LP-1–derived RNA.

DCs were generated from adherent PBMNCs of an HLA-A3/24, B62/50, Cw9/6 (panel A) and an HLA-A3/24, B35/60, Cw3/4 (panel B) buffy coat in RP-10 medium supplemented with GM-CSF and IL-4 and electroporated on day 6 with total RNA of the LP-1 cell line. After a 24-hour culture with TNF-α as maturation stimulus, the transfected DCs were used for the induction of CTLs. At 5 days after the first restimulation, the cytolytic activity of the cells was determined in a standard⁵¹Cr-release assay, with the use of the myeloma cell lines LP-1 (HLA-A3/24, B7/18, Cw7,−) and U266 (HLA-A2/3, B7/40, Cw3/7); the breast cancer cell line MCF-7 (HLA-A2,−, B18/44, Cw5,−); the ovarian cancer cell line SK-OV-3 (HLA-A3/68, B18/35); the renal cancer cell line A498 (HLA-A2,−, B8,−); and the EBV-immortalized B-cell line Croft (HLA-A2,−, B7/8, Cw7,−) (loaded with the HIV peptide) as targets.