Figure 5.

Enhanced tumor infiltration of adoptively transferred anti–VEGFR-2 CAR and IL-12–cotransduced T cells in mice bearing established B16-F10 tumor. C57BL/6 mice–bearing B16 melanoma tumor were sublethally irradiated with 5 Gy TBI and treated with 1 × 10⁶ Ly5.1⁺ syngeneic T cells transduced with anti–VEGFR-2 CAR or empty vector alone or either of these vectors cotransduced with NFAT-IL12 vector. Control group received no treatment. Each group included minimum of 14 mice. A, serial, blinded tumor measurements were obtained from 5 mice per group and the products of perpendicular diameters were plotted ± SEM. Tumors and spleens of 3 mice from each group were excised at different time points post-therapy and processed to obtain single-cell suspensions and analyzed by flow cytometry. Percentage of the Ly5.1⁺ lymphocytes was determined in total viable fraction of the cell preparations by flow cytometry. Absolute numbers of Ly5.1⁺ cells were determined by multiplying the percentage of Ly5.1⁺ cells by the total number of viable cells. B, representative FACS data from single-cell preparations of spleen and tumor tissues from 1 mouse in each group obtained on day 6 post-ACT showing the percentage Ly5.1⁺ cells gated in the total viable cell population. C, pooled data obtained from 3 mice from each group collected at indicated time points post-ACT showing the percentage and total number of Ly5.1⁺ cells in spleen and tumor tissues. Data represented as mean ± SEM.