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. 2021 Dec 27;14(1):106. doi: 10.3390/cancers14010106

Figure 2.

Figure 2

Identification of neoantigens: (a) YTN2-reactive CD8+ T cell lines were established from the spleens of mice that had rejected YTN2 tumors. They responded to both YTN2 and YTN16 tumor cells and produced IFN-γ. (b) YTN16-reactive CD8+ T cell lines were established from splenocytes of mice that had rejected YTN16 following treatment with anti-PD-1 and/or anti-CTLA-4 mAbs. IFN-γ production by YTN16-reactive CD8+ T cells stimulated with YTN16 or YTN2 was quantified. Venn diagrams indicate (c) missense mutations identified by WES, (d) expressed mutations filtered by RNA-Seq data (FPKM ≥ 30 and RNA VAF ≥ 0.04), and (e) candidate neoepitopes of YTN2 and YTN16 cells. (f) YTN16-reactive CD8+ T cell lines were stimulated with 11 candidate peptides. IFN-γ production in the culture supernatant was evaluated by ELISA. Data are based on 3 independently established YTN16-reactive CD8+ T cell lines. A dotted line indicates mean + 2 SD of IFN-γ production by the unstimulated cells.