Figure 1.

Treatment of CT26 bearing mice with a TLR7 agonist induces a discrete neoepitope – specific T-cell response against mutated Smc3. A: Design of the peptide matrix encoding all 628 transcribed non synonymous single nucleotide variants (nsSNVs) of CT26. B: Splenocytes were isolated from CT26-WT tumor bearing mice (n = 3, 28 days after tumor inoculation, mean tumor size ~800mm³). 5 × 10⁵ CD4 depleted cells per well were tested for recognition of matrix peptides or 5 × 10⁴ CT26-WT cells in an IFNγ ELISpot. C-D: CT26-WT (C) or CT26-gp70KO (D) tumor bearing mice (n = 5) were treated repetitively with SC1, an TLR7 agonist injected into the tumor starting at day 14 (tumor size ~50 mm³). T-cell responses were analyzed by ELISpot on day 31 as described above. E-F: Splenocytes from TLR7 treated CT26-WT (E) or CT26-gp70KO (F) tumor bearing mice were tested for recognition of Smc3 and gp70 AH1 peptides at 0.4 µg/ml (same concentration as compared to the peptide matrix) or 2 µg/ml as well as CT26-WT or CT26-gp70KO cells by IFNγ ELISpot. Mean + s.e.m. of duplicates is shown.