Fig. 6: Validation of neoantigen-stimulated T cell responses from prostate cancer patients.

a, Schematic of the validation experiment. Somatic mutations were identified by the WES of surgically resected prostate tissues and matched normal cells (PBMCs), and their expression was confirmed by tumour RNA-seq. Candidate immunogenic peptides were selected and validated based on the statistics of binding prediction by PISTE. b, In vitro detection of T cell responses for PBMCs stimulated with individual neoantigens by IFN-γ ELISA. c, IFN-γ production on CD8+ T cells against selected peptides was detected by flow cytometry for patient PCA03 and patient PCA06. Percentages shown in the density plots are frequencies of reactive IFN-γ+ cells as a proportion of all the CD8+ T cells. P value was determined via one-way analysis of variance with Dunnett’s multiple comparisons test. n = 3 repeated technical measurements; error bars show the standard error of the mean. The dashed line indicates the baseline for the identification of positive reactions (Methods). SSC-H, side scatter height. Panel a created with BioRender.com.