Extended Data Fig. 10. Combinatorial signal enhanced CD19CAR-iCAR-TCTL anti-tumor function.
a Surface antigen profiles of CD19BBz-iCAR-TCTL and CD19BBz-DGK-dKO-iCAR-TCTL-mbIL15tg. CD19BBz-iCAR-TCTL and CD19BBz-DGK-dKO-iCAR-TCTL-mbIL15tg were evaluated using surface antigen expression related to T/NK linage, naïve/memory, and exhaustion phenotype. b-c. Detection of phosphorylated ERK (pERK) in CD19BBz-iCAR-TCTL and CD19BBz-DGK-dKO-iCAR-TCTL-mbIL15tg 60 minutes after co-culturing with irradiated Nalm6. Two-sided Student’s t test. d. In vitro cytotoxic assays of CD19BBz-iCAR-TCTL and CD19BBz-DGK-dKO-iCAR-TCTL-mbIL15tg against Nalm6. n = 4, mean ± SEM. e. Cytokine production of CD19BBz-iCAR-TCTL and CD19BBz-DGK-dKO-iCAR-TCTL-mbIL15tg 48 h after co-culturing with irradiated Nalm6 (n = 4, mean ± SEM). Two-sided Student’s t test. f–i. Therapeutic efficacy of CD19CAR-iCAR-TCTL with in the Nalm6 systemic tumor model. Treatment schedule (f). NSG mice were injected intravenously with 5 × 105 Nalm6 cells 4 days before the treatment. In total, 1 × 107 CD19BBz-iCAR-TCTL and CD19BBz-DGK-dKO-iCAR-TCTL-mbIL15tg were injected intravenously on days 4. In vivo bioluminescence imaging of the injected T cells in NSG mice treated with CD19BBz-iCAR-TCTL, CD19BBz-DGK-dKO-iCAR-TCTL-mbIL15tg (g). Total flux (photons/s) of the injected CAR-T cells was quantified at the indicated time points (n = 4 or 5, mean ± SEM). Two-way ANOVA Turkey’s multiple comparison (h) mice blood cells were collected on day 23. Nalm6 (hCD45 + CD19 + ) cells were analyzed by flow cytometry. One-way ANOVA with Tukey’s multiple comparisons test (i).