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.