Fig. 2: Neoantigen-reactive CD8⁺ T cells recognise cognate peptide in vitro.

a Table detailing attributes of tumour-reactive T cell receptors (TCRs), including neoantigen-reactive (top) and tumour-associated antigen-reactive (TAA-reactive) (bottom) TCRs isolated upon vaccination. b Neoantigen-reactive CD8⁺ T cell responses were induced by vaccination; isolated tetramer⁺ T cells were stimulated for 6 h with (varying concentrations of) cognate peptide and αCD28 and subsequently analysed via intracellular staining (ICS) flow cytometry for IFNγ expression. c Transgenic (tg)TCR CD8⁺ T cells were co-incubated with varying concentrations of cognate mutant (MT) peptide or wild type (WT) peptide (x-axis) and αCD28 for 6 h; intracellular staining (ICS) was performed subsequently. Percentage of T cells expressing IFNγ of a parent CD3⁺CD8⁺ population is shown on y-axis, normalised to maximum IFNγ expression. MT peptide values are shown as clear circles, and WT as filled, black circles. Trp2-reactive TCR 180CC6 only recognises a WT peptide (shown with clear circle). d Neoantigen-reactive TCR half-maximal (EC₅₀) cytokine production concentration (top). TgTCR CD8⁺ T cells expressing the indicated TCR were stimulated, as described in (c), with titrated mutant (MT) or wild-type (WT) peptide and IFNγ production was measured by ICS. Symbols indicate median of biologic replicates (n = 3/condition), ±95% CI. Dashed horizontal line indicates mean half-maximal response (EC₅₀) for tested neoAg TCR. (Bottom) Ratio of WT/MT EC₅₀ as log₁₀ fold change. Source data are provided as a Source Data file.