Fig. 1: Collecting and expanding TIL from human colorectal cancer specimens.

a Processing workflow of clinical specimens. b The T-cell population in PBMC and TIL. Left: representative flow cytometry plots of PBMC and TIL. Upper data were gated on live cells. Lower data were gated on CD3⁺ live cells. Right: quantification of CD3⁺ cells or CTL frequency in PBMC and TIL. ****P < 0.0001, two-tailed unpaired t test. Dots represent data from individual cases. Means are shown; error bars represent SD. c Number of bulk TIL and TI-CTL from an ~5 mm³ tumor block. Dots represent data from individual cases. Medians are shown; error bars represent interquartile ranges. d Expansion data of TI-CTL with cytokine cocktails. Expansion folds of TI-CTL were compared on day 14. *P < 0.05, paired nonparametric analysis (Friedman test followed by Dunn’s multiple comparisons test). Dots represent data from eight individual cases. Medians are shown; error bars represent interquartile ranges. e The TCR Vβ repertoire analysis of TI-CTL based on flow cytometry. Left: representative clonogram from both pre- and post-cytokine expansion TI-CTL for C-T-10 MMR-D. Arrows indicate Vβ subtypes containing tumor-specific populations. Right: dots indicate the number of detected Vβ subtypes from individual cases. NS, not significant, two-tailed paired t test. Means are shown; error bars represent SD.