Figure 1.
Generation of EBNA1-specific T cells. (A) T cells were generated from HLA-B8–expressing PBMCs of donor M after in vitro stimulation with synthetic peptides from EBNA1. 1.5 × 105 cells per well of PBMCs were used to generate EBNA1-P518–530 peptide–specific T cells. Peptides other than those used for repeated stimulations served as negative controls. For T cell recognition assays, peptides were pulsed onto 1359mel target cells and cocultured with T cells overnight. Cytokine release assays were performed as previously described (reference 11). (B) T cell recognition (T cell line M3-W1 from donor M) of 1359mel cells pulsed with EBNA1-P518–530 peptide was specifically inhibited by antibody against MHC class I molecules. T cell recognition assays were performed at an E/T ratio of 1:1. All results are expressed as IFN-γ release in picogram/milliliter and are the averages of duplicate values. All antibodies were used at a final concentration of 20 μg/ml each.