Figure 4. RNA pentatope vaccines with mutations selected for in silico predicted favourable MHC class II binding and abundant expression confer potent antitumour control.

a, Comparison of median MHC II binding scores of immunogenic (Response) and non-immunogenic (No response) mutated 27mers. b, Highly expressed mutations were selected with (‘ME’) or without (‘E’) considering MHC class II binding score. Ten mutations (two pentatopes) per category were used for vaccination of CT26-Luc tumour-bearing mice (n = 10). Tumour growth, area under the curve (AUC) at day 40 and ink-treated lungs are shown. c, Mice (n = 5) were analysed for T-cell responses against the RNA pentatopes via ELISpot (mean ± s.e.m. subtracted by an irrelevant RNA control). d, CT26 tumour nodules per lung of untreated mice or mice (n = 10) injected with irrelevant or P_ME (±CD8 depletion or CD40L blocking) RNA. e, T-cell responses against gp70_423–431 (gp70-AH1) determined via ELISpot in blood (pooled from 5 mice at day 20) and spleen (n = 5). Background (medium control) subtracted mean ± s.e.m. shown. f, Genomic, expressed and predictively presented (HLA-DRB1, IEDB rank <10) non-synonymous single nucleotide variations (nsSNVs) derived from human cancer samples (TCGA). SKCM, skin cutaneous melanoma; COAD, colon adenocarcinoma; BRCA, breast invasive carcinoma.