Abstract: Objective To construct recombinant adenovirus expressing both simulated peptide of blood group B antigen P1/Fas and CCL19 by pAdEasy-1 packaging system,and test their expression. Methods We constructed pShuttle-CMV-P1/Fas-CCL19 by adenovirus Shuttle plasmid pShuttle-CMV and pAdEasy-1.Then，we transfected pAd-CMV-P1/Fas-CCL19 to HEK293A cells and obtained recombinant Ad-P1/Fas-CCL19 particles which can infect 4T1 cells.At last we detected mRNA and protein expression of P1/Fas-CCL19 in 4T1 cells.4T1 tumor-bearing mice model immuned by the B red blood cell was established successfully,and injected with recombinant adenovirus Ad-P1/Fas-CCL19,empty adenovirus,saline into the tumors,respectively.Then,tumor changes and survival period of the three groups were compared. Results P1/Fas-CCL19 can be effectively expressed in 4T1 cells on both mRNA and protein levels.Intratumor injection of recombinant adenovirus Ad-P1/Fas-CCL19 group inhibited 4T1 tumor growth significantly compared with intratumoral injection of adenovirus,empty saline,but survival time has no obvious difference among the three groups. Conclusion P1/Fas-CCL19 recombinant adenovirus vector was successfully constructed,experiments proved its stable expression and efficacy in vivo,in vitro，which make a foundation for further clinical research.