Figure 8.
Imaging of Anti-tumor Activity and Persistence of αvβ6-Re-targeted CAR T Cells in SCID Beige Mice with Panc0403 PDAC Xenografts
(A) T cells were co-transduced with retroviral vectors encoding for rluc/GFP together with A20-28z/4αβ or the truncated control, A20-Tr/4αβ. After culture for 12 days in IL-4, cells were analyzed by flow cytometry. 9e10 detects a myc epitope tag in the CAR ectodomain. SSC, side scatter. Quadrants were set using untransduced T cells cultured in IL-2. (B) Mice were injected i.p. with 2 × 106 Panc0403-ffluc cells, and tumors were allowed to establish for 14 days before i.p. treatment with 10 × 106 of the indicated gene-modified T cells or PBS as control (arrow). Bioluminescence imaging using d-luciferin (substrate for ffluc) was used to monitor tumor status. Data show the mean ± SEM of tumor-derived total flux (n = 5 mice/group). (C) Mice were weighed weekly to assess toxicity of CAR T cells. Data show the mean ± SEM weight of n = 5 mice/group. To image T cells, BLI was performed after the administration of coelenterazine at the indicated intervals, commencing 1 hr after T cell injection. Images of individual mice (D) and pooled BLI data (E; mean ± SEM, n = 5) are shown.