Figure 6.

Comparison of Therapeutic Effect by Single Epitope or Multiple Epitopes of Neoantigen

(A) Tumor therapeutic model. C57BL/6J mice were s.c. injected with MC38 tumor cells and treated with neoantigen single-peptide- or multipeptide-pulsed DCs at a week later and again with the relevant peptide together with poly(I:C) and anti-CD40 Ab. Tumor growth was monitored at indicated time points. Tumor growth curve in each group of mice was plotted. As shown by the red line, tumor volume <400 mm³ at day 25 was consider as a partial response. Data are pooled from five independent experiments (n = 11–13).

(B and C) Analysis of TIL in mice treated with multipeptides of neoantigens. TILs were stimulated by peptides in the presence of anti-CD28 and brefeldin. (B) The percentages of IFN-γ single-producing T cells (left), IFN-γ⁺TNF-α⁺-producing T cells (middle), and IFN-γ⁺TNF-α⁺IL-2⁺-producing T cells (right) were plotted. Data are pooled from three independent experiments (n = 6). (C) Representative flow cytometry analysis of CD8⁺ TILs shows IFN-γ and TNF-α.

(D) Therapeutic effect by neoantigen-pulsed DCs. MC38 cells were administered s.c. One week later, these mice were treated with neoantigen-pulsed DCs twice at 1-week intervals. Anti-PD-1 Ab was injected intraperitoneally (i.p.) at day 7, 10, and 13 (n = 10 and 11/group). Data are pooled from four independent experiments, and the values represent mean ± SEM. Data were analyzed by one-way ANOVA followed by Turkey's multiple comparison test. ∗p < 0.05.