Objective:To discuss the cytotoxic activity of CTL,stimulated by adenovirus containing the universal DR-restricted Th helper epitope(PADRE) combined with HLA-A1 and HLA-A2 restricted epitopes from mucin4 gene(PADRE/MUC4) infected DCs. Methods:Recombinant adenovirus with PADRE/MUC4 gene was prepared,DCs of HLA-A2 positive donors were induced from PBMC in vitro. PBMC primed with adenovirus infected DC for 3 weeks,the specific cytotoxic activity of CTL was detected by standard 4 h Cr51 release assay and Elispot method. Results:Lymphocytes primed with pAd-CMV-PADRE/MUC4 caused potent cytotoxic responses. By contrast,lymphocytes primed with a GFP expressing adenovirus or mock-infected DCs had no cytotoxic effect. Conclusion:Infection of DCs with an adenovirus encoding PADRE combined with epitopes from mucin4 gene may be one possible strategy for immunotherapy of MUC4-expressing tumors.