It remains a top research priority to develop immunotherapeutic approaches to induce potent antigen-specific immune responses against tumors. However, in spite of some promising results, most strategies are ineffective because they generate low numbers of tumor-reactive cytotoxic T lymphocytes (CTLs). Here we designed a strategy to enhance antigen-specific immune response via administering sulfosuccinimidy1-4-[N-maleimidomethyl] cyclohexane-1-carboxylate (sulfo-SMCC)-conjugated melanoma tumor antigen GP100(25-33) peptide-coupled syngeneic spleen cells in a mouse model of melanoma. We found that infusion of GP100(25-33) peptide-coupled spleen cells significantly attenuated the growth of melanoma in prophylactic and therapeutic immunizations. Consistent with these findings, the adoptive transfer of spleen cells from immunized mice to naive syngeneic mice was able to transfer anti-tumor effect, suggesting that GP100(25-33) peptide-specific immune response was induced. Further studies showed that, CDS+ T cell proliferation and the frequency of interferon (IFN)-gamma-producing CD8+ T cells upon ex vivo stimulation by GP100(25-33) were significantly increased compared to control groups. Tumor antigen, GP100(25-33) specific immune response was also confirmed by ELISpot and GP100-tetramer assays. This approach is simple, easy-handled, and efficiently delivering antigens to lymphoid tissues. Our study offers an opportunity for clinically translating this approach into tumor immunotherapy. (C) 2015 Elsevier Inc. All rights reserved.