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Dendritic cell-based cancer vaccines(DC vaccines)have been proved efficient and safe in immunotherapy of various cancers,including melanoma,ovarian and prostate cancer.However,the clin-ical responses were not always satisfied.Here we proposed a novel strategy to prepare DC vaccines.In the present study,a fusion protein SNU containing a secretin-penetratin(SecPen)peptide,NY-ESO-1 and ubiquitin was designed and expressed.To establish the DC vaccine(DC-SNU),the mouse bone marrow-derived DCs(BMDCs)were isolated,pulsed with SNU and maturated with cytokine cocktail.Then pe-ripheral blood mononuclear cells(PBMCs)from C57BL/6 mice inoculated intraperitoneally with DC-SNU were separated and cocultured with MC38/MC38NY-ESO-1 tumor cells or DC vaccines.The results show that SNU was successfully expressed.This strategy made NY-ESO-1 entering cytoplasm of BMDCs more efficiently and degraded mainly by proteasome.As we expected,mature BMDCs expressed higher CD40,CD80 and CD86 than immature BMDCs.Thus,the PBMCs released more IFN-γ and TNF-αwhen stimulated with DC-SNU in vitro again.What's more,the PBMCs induced stronger and specific cytotoxicity towards MC38NY-ESO-1 tumor cells.Given the above,it demonstrated that DC-SNU loaded with SecPen and ubiquitin-fused NY-ESO-1 could elicit stronger and specific T cell immune responses.This strategy can be used as a platform for DC vaccine preparation and applied to various cancers treat-ment.