Immunotherapy of tumor-bearing mice with DCs transfected with angiogenesis-associated and tumor antigens. (A-B) B16/F10.9 melanoma model: C57BL/6 mice were implanted subcutaneously with 1 × 104 B16/F10.9 tumor cells and 3 days later immunized with mRNA-transfected DCs 3 times at 7-day intervals. Mice were immunized with 3 × 105 DCs for each antigen for a combined 6 × 105 DCs per mouse. Data in panel A are 18 days after tumor implantation and in panel B are 25 days after tumor implantation. Columns represent mean lung weight and dots represent individual lung weight (7 mice per group). (A) Relative to the control group (PBS), P values were less than .05 for all groups except actin (P = .4206). The overall significance of the study as determined by the Kruskal-Wallis test was P < .0001. (B) Relative to the control group (PBS), P values were less than .05 for all groups except actin (P = .4206). The overall significance of the study as determined by the Kruskal-Wallis test was P = .0011. (C-D) Figure shows time to appearance of palpable tumors in the experiment shown in panel B. The log-rank test (Mantel-Haenszel test) was used to determine the differences between individual groups. (C) Relative to the combination group (TRP-2 + VEGFR-2), P values were .03 and .002 for groups immunized with TRP-2 + actin and VEGFR-2 + actin, respectively. The median time to tumor onset for TRP-2 + actin was 18 days, VEGFR-2 + actin was 15 days, and TRP-2 + VEGFR-2 was 25 days. (D) Relative to the combination group (VEGF + TRP-2), P values were .03 and .02 for groups immunized with VEGF + actin and TRP-2 + actin, respectively. The median time to tumor onset for TRP-2 + actin was 18 days, VEGF + actin was 18 days, and VEGF + TRP-2 was 37 days.
