Figure 6

The processing efficacy of the gp100^mel–derived spliced epitopes is comparable to that of the non-spliced _M210gp100^mel_209–217 and NY-ESO-1_157–165 epitopes.

(A–E) The IFN-γ-release by specific CD8⁺ T cell clones exposed to HLA-A*03:01/-A*02:01/-A*32:01⁺ HeLa cells (E:T = 1:1) pulsed with different concentrations of the synthetic peptides (upper panels) or HLA-A*03:01/-A*02:01/-A*32:01⁺ HeLa cells transfected with the target antigen (lower panels) is shown for the epitopes _M210gp100^mel_209–217 (A), NY-ESO-1_157–165 (B), gp100^mel_{40–42/47–52} (C), gp100^mel_{47–52/40–42} (D) or gp100^mel_{195–202/192} (E). The data are the mean and bars the SD of three replicates of a representative assay of independent experiments (n = 2–6). (F) We here report a marker of the amount of epitope presented on the cell surface of HLA-A*03:01/-A*02:01/-A*32:01⁺ HeLa cells transfected with the target antigen. By titrations (upper panels in A–E) we calculated the amount of synthetic peptide pulsed on to HeLa cells, which triggered the release of IFN-γ amount corresponding to the IFN-γ released by the specific CD8⁺ T cell clones exposed to HLA-A*03:01/-A*02:01/-A*32:01⁺ HeLa cells transfected with the target antigen and cultivated in an E:T ratio = 4:1 (see lower panels in A–E). Values are the means and bars the SEM of independent experiments (n = 2–7; each experiment with three independent replicates). Significant differences in the means are marked and the corresponding by p values are reported (we applied the Mann-Whitney test with Bonferroni correction for multiple comparison). The IFN-γ-release in the medium was measured by ELISA after 16 hrs co-culture.