Figure 2. Antitumor activity of vaccination with predicted tumor neoantigen (TNA).

(A) Scheme of immunization. Two doses of peptides emulsified in Freund’s adjuvant were subcutaneously (s.c.) injected, separated by 14 days. 14 days after the last dose the B16 F10 melanoma cells were injected s.c. in the mid-right flank of C57BL/6J host mice and the tumor size over time was analyzed using a dial caliper. (B–I) Tumor growth on non-vaccinated (B) or vaccinated mice with the indicated peptide (C–I) (or peptide mix (H)), monitored every 1–3 days. Each line corresponds to the tumor size in one animal. The median of the tumors in non-vaccinated (phosphate-buffered saline [PBS]) and vaccinated with Trp2 (positive control) are shown as dashed lines in gray and green color, respectively. Mice with tumors≥900 mm³ were sacrificed. Following the rules of our ethical committee, animals presenting ulcers were also sacrificed. (J) Percentage of animals showing tumors with a volume≤100 mm³ 15 days after implantation. 4 mice/group were used.