Fig. 4

Anti-tumor activity of CD7 KO-CD7 CAR-iNK cells against primary T-ALL cells. A Experimental design for the T-ALL patient tumor-killing assay by hPSC-derived CD7 KO-CD7 CAR-iNK cells. Mononuclear cells (MNCs) were first isolated from the bone marrow of patients with T-ALL and transplanted into B-NDG immunodeficient mice (1 × 10⁶/mouse) via tail vein injection. Four weeks after transplantation, the proportion of primary tumor cells in the PB of B-NDG mice was monitored weekly using flow cytometry. Once the proportion of huCD45⁺CD7⁺ primary tumor cells exceeded 80% (approximately 6-9 weeks post-transplantation), the spleens of the B-NDG recipient mice were harvested and processed into single-cell suspensions. The resulting splenocytes were then cocultured with Ctrl-iNK or CD7 KO-CD7 CAR-iNK cells for tumor cytotoxicity assays. B Surface expression of CD7 in the splenic cells from B-NDG recipient mice measured by flow cytometry. C-E Cytotoxicity analysis of Ctrl-iNK or CD7 KO-CD7 CAR-iNK cells against T-ALL cells isolated from patient 1 (C), patient 2 (D), and patient 3 (E) at the indicated E:T ratios after 6-h incubation. Data are represented as mean ± SD (n = 4). Two-way ANOVA and Mann-Whitney U test were used for statistics. ***p < 0.001. F–H Cytotoxicity analysis of Ctrl-iNK or CD7 KO-CD7 CAR-iNK cells against T-ALL cells isolated from patient 1 (F), patient 2 (G), and patient 3 (H) at the E:T ratio of 1:2 after 3 h-, 6 h-, 9 h-, and 12 h- incubation. Data are represented as mean ± SD (n = 4). Two-way ANOVA and Mann-Whitney U test were used for statistics. ***p < 0.001