Fig. 5. In vitro and in vivo functions of WT1-TCR-transduced HLA-homo iCD8αβ T-cells.

a Fold proliferation of WT1-TCR iCD8αβ T-cells in an optimized protocol as shown (top) for four sequential rounds of stimulations. Data are representative of two independent experiments. b Flow cytometry plots of proliferated WT1-iPSC CD8 SP T-cells representing the expression levels of tetramer and naive/memory T-cell markers (n = 3 independent experiments). c In vitro cytotoxicity assay of proliferated WT1-TCR iCD8αβ T-cells measured by DELFIA assay using LCL cells loaded with specific peptides as target cells (left) and NCI-H226 (right). Data are representative of two independent experiments. d Intracellular cytokine production of WT1-TCR iCD8αβ T-cells after 6 h stimulation with PMA/Ionomycin. Data are representative of two independent experiments. e, f In vivo anti-tumor activity of WT1-TCR iCD8αβ T-cells. NSG mice were intraperitoneally inoculated with NCI-H226-expressing luciferase, treated 4 times weekly with HBSS or WT1-TCR iCD8αβ T-cells, and monitored for (e) tumor volume and (f) survival rate (n = 5 animals each). *P = 0.0016. Data represent mean ± SEM. ^*P < 0.05; ^**P < 0.01 (Welch’s two samples t-test (two-tailed) (e); log-rank test (two-tailed) (f)).