Fig. 2.

Identification and selection of tumor associated antigens and neoantigens. (A) Heatmap of mRNA expression of tumor associated antigens (TAAs) in the patient's tumor sample. The mRNAs of the indicated TAAs in tumor samples from resection were assayed by quantitative PCR (qPCR) in triplicate, and the expression levels of the measured genes were expressed as the mean fold changes. (B) Procedure for neoantigen identification and selection. The number of identified candidates is indicated for each step. (C) In vitro T-cell responses to synthesized neoantigen and wild-type peptides. Peripheral blood mononuclear cells (PBMCs) were stimulated with autologous dendritic cell (DC) pulsed with antigen peptides for 12 days and restimulated for 6 h. T cells were stained for intracellular expression of tumor necrosis factor alpha (TNF-α) and interferon-gamma (IFN-γ), and percentages of cytokine-expressing CD4⁺ or CD8⁺ T cells are shown. NGS: next generation sequencing.