CCR4-CAR (KW_L2H) induces comparable antitumor efficacy to CD19-CAR in NSG mice engrafted with HH expressing CD19 (HH-CBG-GFP-t19). (A) Schematic representation of the experiment. NSG mice were intravenously inoculated with 1 × 106 HH cells expressing luciferase and truncated CD19 (HH-CGB-GFP-t19). Engrafted tumors were intravenously treated with either 2 × 106 CCR4-CAR–positive T cells or UTD T cells 7 days after tumor cell inoculation. (B) Overall kinetics of the tumor burden by BLI. The dashed line represents the background level of photons determined by imaging tumor-free mice. Dots and bars represent the mean and SEM (n = 5 for the UTD group and n = 7 for the other groups). The data are representative of 2 experiments from 2 donors. (C) T-cell counts in peripheral blood on days 7 and 14. Peripheral blood was analyzed for T-cell counts on days 7 and 14 by FCM using counting beads. Bars represent the mean and SEM (n = 5 for the UTD group and n = 7 for the other groups). The data are representative of 2 experiments from 2 donors. (D) Cytokine levels of serum on day 6. Serum was collected from mouse whole blood at day 6 using the same experimental schedule as described in panels A-C. Cytokines were analyzed using high-sensitivity LUMINEX assay. Bars represent the mean and SEM (n = 4 for the UTD group and n = 5 for the other groups). ∗P < .05; ∗∗P < .01; ∗∗∗∗P < .0001 by 1-way ANOVA with Tukey’s post hoc test. ns, not significant.
