Objective To investigate the effects of combination palmitoylated peptide and poly IC on the CD8⁺T cells specific for melanoma. Methods 15 C57BL/6 mice were randomly divided into five groups: controls group(only normal saline wasinjected), peptide group(gp100_25-33, melanoma specific CD8⁺T cell epitope), peptide plus poly IC group, palmitoylated peptide group and palmitoylated peptide plus poly IC group. After vaccination via intramuscular injection, CD8⁺T cells from peripheral blood and spleens of immunized mice were harvested for. Analyzing the number of CD8⁺T cells specific for melanoma by flow cytometry, and detecting the recognition of B16F10 melanoma cells ex vivo by Elisa and Elispot. Further anti-tumor effects were evaluated in 15 melanoma bearing mice after vaccination. Results Comparing with peptide plus poly IC group, palmitoylated peptide plus poly IC significantly increased the number of gp100_25-33 specific CD8⁺T cellsin peripheral blood[(23.53±5.398)% vs(1.158±0.239)%,P=0.014]. Furthermore, endogenous CD8⁺T cells in duced by vaccine recognized B16F10 melanoma cells ex vivo, and dramatically inhibited tumor growth in vivo. Conclusion Combination of palmitoylated peptide and poly IC can induce strong CD8⁺T cell response, resulting in significant tumor growth inhibition.