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. 2020 Jun 4;23(6):101238. doi: 10.1016/j.isci.2020.101238

Figure 4.

Figure 4

Bioactive Neoantigen Epitopes

Analysis of neoantigen-specific T cells in vaccinated mice. C57BL/6J mice were administered intravenously (i.v.) with neoantigen peptide (Zbtb40, Dpagt1, and Cry1) together with poly(I:C) and anti-CD40 Ab.

(A) One week later, neoantigen-specific T cell activation in these mice was assessed by intracellular staining for IFN-γ and TNF-α production (i) and upregulation of PD-1 and CD137 (ii) after culturing with or without each peptide for 6 h (i) or 24 h (ii).

(B) As shown in (A), neoantigen-specific T cell proliferating responses in these mice were assessed by multimer-APC, CD8a-FITC, and TCR-β-PE. Representative flow cytometry analysis (i) and each value was plotted (ii).

(C) Neoantigen-specific T cell response was quantified. Whole splenocytes (ELISA) or isolated CD8+ T cells (ELISPOT) from mice given each neoantigen were cultured with each neoantigen peptide or non-mutated relevant peptide for 24 h and assessed for IFN-γ production. Data are pooled from two independent experiments, and the values represent mean ± SD. Circles and dot lines link respective mice.