Identification of CD8-restricted T-cell epitopes
(A) IFN-γ production by T-cells in response to PRAME peptide pool stimulation. (B)(C) T-cell responses to single PRAME peptides, present in the mini-pools, were evaluated by IFN-γ ELISpot assay. (D) HLA-I-restricted epitopes (36, 37, 43, and 44) were determined by measuring IFN-γ and TNF-α release by CD8+ T-cells. (E) CD4+ T-cells showed no specificity for HLA-I-restricted epitopes (36, 37, 43, 44). (F) The minimal 9-mer peptides FPEPEAAQP and VEVLVDLFL were determined by IFN-γ ELISpot assay. (G) FPEPEAAQP HLA restriction was confirmed using anti-HLA-B*35 antibody.