Fig. 8. HIP CAR T cells quickly clear tumor cells re-injected after 83 days in fully immunocompetent allogeneic humanized mice.

a Humanized mice were injected with 1 × 10⁶ Luc⁺ Nalm6 cells and received 7 × 10⁶ allo CAR T cells or HIP CAR T cells on day 3 (one experiment). Then, they received another injection of 1 × 10⁶ Luc⁺ Nalm6 cells on day 83. Spleen and bone marrow were taken after 95 days. b, c Human immune cell reconstitution is shown for all 5 mice in the allo CAR T group (b) and the HIP CAR T group (c). Individual values are shown and mean ± SD. d BLI images show the tumor burden for all mice in this study. e–g Graphs show BLI signals for animals in the HIP CAR T (e) and allo CAR T cell groups (f), and the Nalm6 only group (g, all single animals are shown). Arrows indicate Nalm6 injections, * indicates outliers used for further analysis. h, i Reconstitution of human immune cells was assessed in the HIP CAR T (h) and the allo CAR T (i) groups after euthanasia. The percentage of hCD45⁺ cells among all CD45⁺ cells and the percentage of hCD3⁺ cells in the hCD45⁺ population was quantified (mean ± SD, n = 5 HIP animals, n = 4 allo animals with maintained hCD3⁺ population and n = 1 allo animal with lost hCD3⁺ population). j, k The percentages of CAR⁺ cells in the bone marrow (j) and spleen (k) were assessed separately for animals that had maintained their hCD3⁺ population (n = 5 in the HIP CAR T group and n = 4 in the allo CAR T group, mean ± SD). The one animal in the allo CAR T group that had lost their hCD3 population is shown separately. Significant differences between the two CAR T cell groups are shown. l The percentage of CD19⁺ cells in the bone marrow was assessed separately for animals that had maintained their hCD3⁺ population (n = 5 in the HIP CAR T group and n = 4 in the allo CAR T group, mean ± SD). The one animal in the allo CAR T group that had lost their hCD3 population is shown separately. m The percentages of the HIP-edited cells in the HIP CAR T cell product before injection (n = 3) and the percentages of HIP-edited cells among the recovered CAR⁺ cell population from the spleen on day 95 (n = 5) are shown (mean ± SD, unpaired, two-tailed Student’s t test). For (h–l): Endpoint values from earlier euthanized mice in the allo CAR T group due to high tumor burden were included in the day 95 graphs to show all values at the individual endpoint. For (j–l): Unpaired, two-tailed Student’s t test was used to compare HIP and allo CAR T cell groups and significant differences are shown.