Figure 2.

Monomeric TCR/CD8-pMHC dissociation rates (k_off) of Melan-A-specific CD8 T-cell clones. (A) Representative wild-type NTAmer-based dissociation curves obtained from Melan-A-specific CD8 T-cell clones ex vivo derived from patients vaccinated with either low peptide/CpG (gray), high CpG (green), or high peptide (blue) dose protocol, after 4 (dashed curves) or 8 (plain curves) vaccinations. (B) k_off values categorized according to the vaccination cohort and the number of vaccine injections (4v vs. 8v). (C,D) Direct comparison of the dissociation rates (k_off) between low peptide/CpG and high CpG (left panel) or high peptide (right panel) dose cohort after 4 vaccine (C) or 8 vaccine (D) injections. (E) EC₅₀ (top panel) and B_max (bottom panel) values from cytolytic assays performed with chromium-labeled Melan-A-negative T2 target cells loaded with graded concentrations of native Melan-A peptide. Direct comparison between low peptide/CpG (in gray) or high peptide (in blue) dose cohort after 4 vaccine injections. (A–E) Data are representative of pooled tumor antigen-specific T-cell clones derived from EM28^pos and EM28^neg and are depicted as box (25th and 75th percentiles) and whisker (10th and 90th percentiles) with the middle line indicating the median. Number of clones n and P-values by Mann-Whitney U test are indicated; ^*P < 0.05, ^**P < 0.01, and ^***P < 0.001. Pp, peptide.