Administration of myotoxins at vaccination sites significantly enhanced idiotype DNA vaccine-induced tumor protection. Ten BALB/c mice per group were injected intramuscularly with 6.8 μg cardiotoxin (A) or 0.1 μg crotoxin (B) followed by intramuscular vaccination at the same site 5 days later with 50 μg plasmid DNA encoding MCP3 chemokine-fused A20 lymphoma-derived idiotype antigen (MCP3-sFv). TLR agonists, including TLR3 agonist Poly I:C (C), TLR4 agonist MPL (D), TLR7 agonist (M001; E), and TLR7/8 agonist (M003; F), were given, respectively, on the next day of vaccination at a dose of 50 μg.^19,20  A total of 3 vaccinations were given with an interval of 14 days. Two weeks after final vaccination, all mice were challenged with a lethal dose of 2 × 10⁵ A20 lymphoma cells by intraperitoneal injection and were followed for survival for 80 days. Control mice were injected with plasmid DNA without candidate adjuvants or with PBS. Survival differences between groups were analyzed by log-rank test. The data shown are from a single experiment, with results presented in multiple panels for clarity.