Fig. 2: Selection of tumor-specific TI-CTL from cytokine-expanded populations.

a Overview of the tumor-specific TI-CTL selection from cytokine-expanded populations. b Cytokine-expanded TI-CTL were co-cultured with autologous cancer spheroids. CD107a⁺ or 4-1BB⁺ populations were sorted and selectively expanded by the PHA/PBMC method. All data were gated on CD45⁺ live cells. c Tumor-reactive TI-CTL were co-cultured with cancer spheroids again with or without HLA class I antibody. Representative data of three independent experiments are shown from C-T-10 MMR-D. All data were gated on CD45⁺ live cells. d Activation of tumor-reactive TI-CTL against cancer spheroids was evaluated on a per Vβ-subtype basis. HLA class Ι blocking antibody was used for the prediction of TCR-HLA class Ι axis dependency for the TI-CTL activation. The arrows designate Vβ subtypes that mainly depend on TCR-HLA class Ι interactions for their activation. Left: data from MMR-D. Representative data of two independent experiments are shown. Middle and right: data from MMR-P. Representative data of three independent experiments for each are shown. e Killing function data of the selected Vβ-based TI-CTL subgroups against autologous spheroids. Representative data of three independent experiments are shown. The ⁵¹Cr release assay was applied for the quantification. Dots represent individual values. Means are connected by lines; error bars represent SD. n = 3 per point.