Fig. 1

Identification of neoantigen-specific T cells in PBMC of epithelial cancer patients. a A representative data from one of four patients showing the gating strategy used for the phenotypic analysis of PD-1-positive (PD-1⁺) peripheral T cells. b PBMC isolated from four epithelial cancer patients were thawed, rested overnight in a cytokine-free T-cell medium and stained with antibodies against CD3, CD45RO, CD62L, and PD-1. PD-1⁺ and PD-1-negative (PD-1⁻) cells were analyzed for the surface expression of the T-cell memory markers, CD45RO and CD62L. Error bars indicate the mean standard deviation between all four patients samples. c PBMC isolated from six epithelial cancer patients were thawed, rested overnight in a cytokine-free T-cell medium and stained with antibodies against CD3, CD45RO, CD62L, CCR7, and CD45RA. Cells were sorted based on their memory subset, washed once with PBS, frozen at −80 °C and send for TCR-Vβ sequencing. Numbers represent the percent of specific TCR-Vβ sequence in each T-cell population. A minus sign (−) was used If no sequence was identified. d Analysis of the productive clonality of TCR-Vβ sequences of each T-cell population isolated in c. (Paired t-test, *** P < 0.001, * P < 0.05) dashed line, mean