GAA-vaccines in combination with i.m. poly-ICLC administrations induce long-term anti-GL261 protective immunity. (A), C57BL/6 mice received s.c. GAA-vaccines with or without i.m. poly-ICLC on days -14 and -7. On day 0, mice received i.c. inoculations of 1 × 105 GL261 cells, with survival subsequently monitored. **P = 0.003 for the mice receiving combination treatments compared with the control group. *P = 0.0521 for the mice receiving GAA-vaccines alone compared with the control group (Log rank test). (B and C), mice that survived for 90 days following GAA-vaccines and i.m. poly-ICLC or GAA-vaccine alone in (A) were re-challenged with 5 × 104 GL261 in the contralateral hemisphere of the brain (n = 3/group). As controls, naïve mice received the same number of GL261 cells. BILs were harvested from the tumor-bearing hemisphere at 7 days after tumor re-challenge, and stained with TC-anti-CD8 and PE-H-2Kb/TRP2180–188-specific tetramer. (B), numbers represent the percentage of CD8+/TRP-2 tetramer+ cells in lymphocyte-gated BILs in each group. (C), numbers of viable CD8+ BILs per mouse.