Panel a shows a series of bioluminescent images depicting tumor progression in mice over time with different treatments: NTD, 19BBz-tEGFR, and 19BBz-Rab5. Panel b shows a line graph displaying the probability of survival of mice over time after tumor implantation, with the x-axis representing days after tumor implanted and the y-axis representing the probability of survival. Panel c shows two flow cytometry plots indicating the percentage of GFP and Nalm6 cells in the peripheral blood of mice treated with 19BBz-tEGFR and 19BBz-Rab5. Panel d shows two scatter plots presenting the percentage of GFP and CD3 positive cells in the peripheral blood of mice treated with 19BBz-tEGFR and 19BBz-Rab5. Panel e shows a series of bioluminescent images depicting tumor progression in mice over time with different treatments: NTD, 19BBz-tEGFR, and 19BBz-Rab5. Panel f shows a line graph displaying the total flux (tumor burden) over time in mice treated with NTD, 19BBz-tEGFR, and 19BBz-Rab5, with the x-axis representing days after tumor implanted and the y-axis representing total flux. Panel g shows a line graph displaying the probability of survival of mice over time after tumor implantation, with the x-axis representing days after tumor implanted and the y-axis representing the probability of survival. Panel h shows an image depicting the physical appearance of tumor tissues isolated from mice treated with 19BBz-tEGFR and 19BBz-Rab5. Panel i shows a scatter plot presenting the weight of tumor tissues isolated from mice treated with 19BBz-tEGFR and 19BBz-Rab5, with the x-axis representing the treatment groups and the y-axis representing tumor weight in grams. Panel j shows two flow cytometry plots indicating the CAR surface expression of tumor-infiltrating CARTs 14 days after infusion with 19BBz-tEGFR and 19BBz-Rab5. Panel k shows a scatter plot presenting the number of CD3 positive and G4S positive cells in tumors after 14 days of CARTs infusion, with the x-axis representing the treatment groups and the y-axis representing the number of cells per gram of tumor. Panel l shows two flow cytometry plots depicting the expansion of CAR positive CD3 positive T cells isolated from tumors after 14 days of CARTs infusion with 19BBz-tEGFR and 19BBz-Rab5. Panel m shows a scatter plot presenting the absolute number of CAR positive CD3 positive T cells isolated from tumors after 14 days of CARTs infusion, with the x-axis representing the treatment groups and the y-axis representing the number of cells per gram of tumor.
Rab5 CARTs exhibit superior in vivo antitumor efficacy. (a) NSG mice were intravenously injected with 1 × 106 Nalm6 tumor cells. 7 days later, mice received 1 × 106 NTDs or the indicated CD19-specific CARTs. Tumor progression was monitored weekly by bioluminescent imaging (BLI). (b) Survival of Nalm6-bearing mice following infusion of the indicated CARTs. Data in a and b are representative of two independent experiments, n = 5 mice for each group. (c and d) Residual tumor cells and T cells in PB from mice in a were analyzed by flow cytometry on day 22 after tumor injection. Data are representative of two independent experiments, n = 5. (e) 1 × 106 K.19.GFP cells were injected s.c. into NSG mice. 7 days later, 1 × 106 of the indicated T cells were infused. Tumor progression was monitored weekly using BLI for tumor-bearing NSG mice treated with the indicated 19BBz CARTs. (f) Tumor burden and statistical analyses of e. Total flux was quantified as the total photon count of each mouse. (g) Survival curve of tumor bearing mice infused with the indicated CARTs. Data in e–g are representative of three independent experiments, n = 5 mice for each group. (h and i) Image (h) and weight (i) of day 14 tumor tissues isolated from mice treated with 19BBz-tEGFR or 19BBz-Rab5 CARTs. Data are representative of two independent experiments, n = 6–7 mice for each group. (j and k) CAR surface expression of tumor-infiltrating CARTs 14 days after infusion. (l and m) CART expansion (l) and absolute number (m) of CAR-positive CD3+ T cells isolated from tumors after 14 days CARTs infusion. Data in j–m are representative of two independent experiments, n = 5-6 mice for each group. Error bars show mean ± SEM. Statistical comparisons were made using unpaired t tests. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001. In b and g, survival curves were compared using the log-rank Mantel–Cox test.