Figure 2.

Identification of mutation-associated neoantigens in LLC1 tumors. (A) C57BL/6 mice (n = 3 or 4) were immunized with 1 × 10⁶ dendritic cells (DC) pulsed with these neoantigen short peptides (Table S1). After two rounds of DC immunization at a 2-week interval, spleen cells from immunized mice were harvested, pooled, and examined for the reactivity to the immunized peptide by ex vivo intracellular cytokine staining (ICS). By ex vivo ICS analysis, 14 peptides (highlighted in red numbers) induced peptide-specific IFN-γ production of CD8⁺ T cells. (B) Spleen cells were also cultured with immunized peptides for 5 days and restimulated with corresponding peptides. After in vitro expansion culture, IFN-γ production was examined by ELISA. Red bars and blue bars represent the results of ex vivo ICS and ELISA after in vitro culture, respectively. Overall, 25 mutated peptides displayed immunogenicity by DC vaccination.