Fig. 4: Peptide immunogenicity assessment via ELISpot assay.

A Schematic listing criteria used for peptide prioritization. B Interferon-gamma (IFN-γ) secretion by PBMCs from healthy donors (HDs) stimulated with selected peptides. Each dot represents an individual peptide (average of technical duplicates). C IFN-γ secretion by PBMCs from ovarian cancer (OVA) patients stimulated with the same peptides; each dot represents a single peptide. In graphs B and C, each dot represents a single peptide (average of technical duplicates). D Interferon gamma secretion upon activation of HD PBMCs stimulated with the selected peptides. Response is deconvoluted at peptide level and each dot represent the response of a different HD. E Interferon gamma secretion upon activation of OVA patients’ PBMCs stimulated with the selected peptides. Response is deconvoluted at peptide level and each dot represent the response of a different patient. F Percent response rate deconvoluted by peptides for both HDs (black) and OVA patients (magenta). Dotted line represents 25% response rate as reference. G Focus on assessment of cancer specific response for peptides with tumor-specific response (4, 8 and 13). Data from peptide 4 and 13 were analyzed using Mann–Whitney test, while data from peptide 8 were analyzed using Wilcoxon signed-rank test (*p value < 0.05) as all HD responses were 0. In all the graphs showing ELISpot data, the dotted line on the y axis represents a response threshold which correspond to 23.3 spots/1*106 PBMCs as from ref. ¹².