CR2/3–CAR-TVSVdisplayed potent antitumor activity and restrained the antigen escape in vivo. (A) Diagram depicting the CAR-T treatment schedule in the mice bearing U87 intracranial tumors. U87 cells were inoculated into the mouse brain at day 0. CAR-T cells were injected at 5 days. Flow cytometry analysis was performed at 14 days. (B) Survival time of NCG mice bearing U87 intracranial xenograft (n = 7 mice). The mice were i.v. treated with VSVΔ51 (2 × 106 PFU), B7H3–CAR-T (106 cells), CR2/3–B7H3–CAR-T(106 cells), B7H3–CAR-TVSV(106 cells), and CR2/3–B7H3–CAR-TVSV(106 cells). (C) H&E staining of mouse brain sections from A was performed at 14 days. Scale bar, 1 cm. (D and E) T cell exhaustion and functional activity of CAR-T cells were analyzed in brain tumor tissue by measuring intracellular expression cytokines granzyme B, TNFα, and IFNγ (D) and the co-expression of the exhaustion markers PD-1, TIM-3, and LAG-3 (E) via flow cytometry (n = 4 biological replicates) at 9 days after CAR-T cell injection. (F) Diagram depicting the generation of antigen heterogeneity glioma model and CAR-T treatment schedule in the mice bearing mixed U87 intracranial tumors (20% B7H3-KO cells + 80% B7H3-WT cells). NCG mice were i.v. injected with 3 × 106 PBMCs at day 0. After 2 days, 20% B7H3-KO– and 80% B7H3-WT–mixed U87 cells (3 × 105 total cells per mouse) were injected into the mouse brain. CAR-T cells, loaded with VSVΔ51 or not, were i.v. injected at 7 days. (G) Kaplan–Meier survival curve of mice bearing U8720% B7H3-KO intracranial tumors (n = 5 mice). (H and I) Monitoring of tumor progression via bioluminescence imaging of luciferase in U87 xenograft mice at the indicated time (n = 5 mice). (H) Representative bioluminescence images of U87 intracranial tumor after treatment. (I) Quantitative analysis of tumor bioluminescence data. (J) Diagram depicting the generation breast cancer model and CAR-T treatment schedule in the mice bearing subcutaneous MDA-MB-231 cells tumors. Breast cancer cells, MDA-MB-231 cells (3 × 106 total cells per mouse), were injected into the mice. CAR-T, loaded with VSVΔ51 or not, were i.v. injected at 7 days. (K) Tumor volume was recorded every 3 days (n = 4 mice). (L) Representative images and weights of tumor xenografts at the end of the experiment (n = 4 mice). Data represent the mean ± SD from one of two independent experiments (B, G, D, E, I, L, and K). Statistics by log-rank Mantel–Cox tests (B and G), one-way ANOVA with Tukey’s multiple comparison test (D, E, I, and L), and two-way ANOVA (K). *, P < 0.05; **, P < 0.01; N.S., no significance.
Figure 7.

CR2/3–CAR-T VSV displayed potent antitumor activity and restrained the antigen escape in vivo. (A) Diagram depicting the CAR-T treatment schedule in the mice bearing U87 intracranial tumors. U87 cells were inoculated into the mouse brain at day 0. CAR-T cells were injected at 5 days. Flow cytometry analysis was performed at 14 days. (B) Survival time of NCG mice bearing U87 intracranial xenograft (n = 7 mice). The mice were i.v. treated with VSVΔ51 (2 × 106 PFU), B7H3–CAR-T (106 cells), CR2/3–B7H3–CAR-T(106 cells), B7H3–CAR-TVSV(106 cells), and CR2/3–B7H3–CAR-TVSV(106 cells). (C) H&E staining of mouse brain sections from A was performed at 14 days. Scale bar, 1 cm. (D and E) T cell exhaustion and functional activity of CAR-T cells were analyzed in brain tumor tissue by measuring intracellular expression cytokines granzyme B, TNFα, and IFNγ (D) and the co-expression of the exhaustion markers PD-1, TIM-3, and LAG-3 (E) via flow cytometry (n = 4 biological replicates) at 9 days after CAR-T cell injection. (F) Diagram depicting the generation of antigen heterogeneity glioma model and CAR-T treatment schedule in the mice bearing mixed U87 intracranial tumors (20% B7H3-KO cells + 80% B7H3-WT cells). NCG mice were i.v. injected with 3 × 106 PBMCs at day 0. After 2 days, 20% B7H3-KO– and 80% B7H3-WT–mixed U87 cells (3 × 105 total cells per mouse) were injected into the mouse brain. CAR-T cells, loaded with VSVΔ51 or not, were i.v. injected at 7 days. (G) Kaplan–Meier survival curve of mice bearing U8720% B7H3-KO intracranial tumors (n = 5 mice). (H and I) Monitoring of tumor progression via bioluminescence imaging of luciferase in U87 xenograft mice at the indicated time (n = 5 mice). (H) Representative bioluminescence images of U87 intracranial tumor after treatment. (I) Quantitative analysis of tumor bioluminescence data. (J) Diagram depicting the generation breast cancer model and CAR-T treatment schedule in the mice bearing subcutaneous MDA-MB-231 cells tumors. Breast cancer cells, MDA-MB-231 cells (3 × 106 total cells per mouse), were injected into the mice. CAR-T, loaded with VSVΔ51 or not, were i.v. injected at 7 days. (K) Tumor volume was recorded every 3 days (n = 4 mice). (L) Representative images and weights of tumor xenografts at the end of the experiment (n = 4 mice). Data represent the mean ± SD from one of two independent experiments (B, G, D, E, I, L, and K). Statistics by log-rank Mantel–Cox tests (B and G), one-way ANOVA with Tukey’s multiple comparison test (D, E, I, and L), and two-way ANOVA (K). *, P < 0.05; **, P < 0.01; N.S., no significance.

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal