Fig. 3.

Cytotoxicity of 5T4_p17-specific TCR transduced CD8⁺ T-cells against T2 pulsed with alanine-substituted 5T4_p17 peptides. a Binding of alanine-substituted 5T4_p17 peptides to HLA-A2 was determined by stabilization of HLA-A2 on the surface of peptide-pulsed T2 cells. CMV_pp65 is an HLA-A2⁺ T-cell epitope from CMV and serves as a positive control. Cell surface HLA-A2 was assessed by immunostaining and flow cytometry. b CD8⁺ T-cells expressing 5T4_p17-specific TCRs were tested for recognition of T2 cells pulsed with the progenitor 5T4_p17 peptide or HLA-A2 binding alanine-substituted variants in a 4-h cytotoxicity assay with a 10:1 E:T