摘要
AIM: To develop an oral DNA vaccine against gastric cancer and evaluate its efficacy in mice.METHODS: The genes of the MG7-Ag mimotope and a universal Th epitope (Pan-DR epitope, PADRE) were included in the PCR primers. By PCR, the fusion gene of the two epitopes was amplified. The fusion gene was confirmed by sequencing and was then cloned into pcDNA3.1(+) plasmid. The pcDNA3.1 (+)-MG7/PADRE was used to transfect an attenuated Salrmonella typhimuriurm.C57BL/6 mice were orally immunized with 1x108 cfu Salrmonella transfectants. Salmonella harboring the empty pcDNA3.1(+) plasmid and phosphate buffer saline (PBS)were used as negative controls. At the 6th week, serum titer of MG7-Ag specific antibody was detected by ELtSA.At the 8th week cellular immunity was detected by an unprimed proliferation test of the spleenocytes by using a [3H]-thymidine incorporation assay. Ehrlich ascites carcinoma cells expressing MG7-Ag were used as a model in tumor challenge assay to evaluate the protective effect of the vaccine.RESULTS: Serum titer of antibody against MG7-Ag was significantly higher in mice immunized with the vaccine than that in control groups (0.841 vs 0.347, P＜0.01; 0.841 vs 0.298,P＜0.01), while in vitro unprimed proliferation assay of the spleenocytes showed no statistical difference between those three groups. Two weeks after tumor challenge, 2 in 7 immunized mice were tumor free, while all the mice in the control groups showed tumor formation.
CONCLUSION: Oral DNA vaccine against the MG7-Ag momitope of gastric cancer is immunogenic. It can induce significant humoral immunity against tumor in mice, and the vaccine has partially protective effects.
AIM:To develop an oral DNA vaccine against gastric cancer and evaluate its efficacy in mice. METHODS:The genes of the MG7-Ag mimotope and a universal Th epitope (Pan-DR epitope,PADRE) were included in the PCR primers.By PCR,the fusion gene of the two epitopes was amplified.The fusion gene was confirmed by sequencing and was then cloned into pcDNA3. 1(+) plasmid.The pcDNA3.1 (+)-MG7/PADRE was used to transfect an attenuated Salmonella typhimurium. C57BL/6 mice were orally immunized with 1×10~8 cfu Salmonella transfectants.Salmonella harboring the empty pcDNA3.1(+) plasmid and phosphate buffer saline (PBS) were used as negative controls.At the 6th week,serum titer of MG7-Ag specific antibody was detected by ELISA. At the 8th week cellular immunity was detected by an unprimed proliferation test of the spleenocytes by using a [~3H]-thymidine incorporation assay.Ehrlich ascites carcinoma cells expressing MG7-Ag were used as a model in tumor challenge assay to evaluate the protective effect of the vaccine. RESULTS:Serum titer of antibody against MG7-Ag was significantly higher in mice immunized with the vaccine than that in control groups (0.841 vs 0.347,P<0.01;0.841 vs 0.298,P<0.01),while in vitro unprimed proliferation assay of the spleenocytes showed no statistical difference between those three groups.Two weeks after tumor challenge,2 in 7 immunized mice were tumor free,while all the mice in the control groups showed tumor formation. CONCLUSION= Oral DNA vaccine against the MG7-Ag momitope of gastric cancer is immunogenic.It can induce significant humoral immunity against tumor in mice,and the vaccine has partially protective effects.
基金
the National Natural Science Foundation of China, No.39870742
