Figure 4.

Expansion of neoepitope peptide-specific T cells from PBMC. Dendritic cells from KADA (A) or ANRU (E) were loaded with their respective TIL-activating and/or tumor-presented peptides and used to stimulate autologous CD8 T cells. The same experiment was performed using three healthy donors for each peptide set (B–D, KADA neoepitopes; F–H, ANRU neoepitopes and MART-1). After 10 days of expansion, the cells were stained with corresponding HLA-A2/peptide dextramers and analyzed by flow cytometry. Dot plots are gated on lymphocytes/singlets/live cells and frequency indicates %dextramer⁺ out of CD8⁺ cells. For ANRU epitopes, the function of the neoepitope specific T cells was assessed by re-stimulation with ANRU tumor cells and evaluated by CD107a staining (I, ANRU; J–L, healthy donors). Only positive stainings are shown. Dot plots are gated on lymphocytes/singlets/live cells/CD8⁺ cells and frequency indicates % dextramer⁺CD07a⁻ or dextramer⁺CD107a⁺ of CD8⁺ cells.