Objective: to transduce the tumor associated antigen gene MAGE-1 and/or IL-12 gene into dendritic cells (DC) and to observe the in vitro cytotoxic effect induced by the genetically modified DC against the human hepato...Objective: to transduce the tumor associated antigen gene MAGE-1 and/or IL-12 gene into dendritic cells (DC) and to observe the in vitro cytotoxic effect induced by the genetically modified DC against the human hepatocellular carcinoma (HCC) cell line SMMC7721. Methods: the MAGE-1 gene was inserted into the retrovirus vector LXSN to construct the recombinant retrovirus LMSN. The monocyte-derived DCs were transfected at appropriate differentiation stage by LMSN and/or a recombinant adenovirus AdmiL-12, which containing murine IL-12 gene. The control groups included retrovirus LXSN transfected, adenovirus AdBGFP transfected and non-transfected DCs. The MAGE-1 gene expression was identified by western blot and the mIL-12 p70 secretion was detected by ELISA assay. The in vitro cytotoxicities against SMMC7721 induced by genetically modified and control groups of DC were tested by MTT assay. Results: The MAGE-1 expression was detected by a monoclonal antibody in DCs tranfected with LMSN but not in control groups. At 16 h, 24 h and 48 h after transfection with AdmIL-12, the concentration of the mIL-12 p70 in the culture medium was 580pg/106 cells, 960pg/106 cells and 1100pg/106 cells respectively. The mIL-12 p70 secretions were not detected in other groups. The lytic activity (as judged by % lysis) induced by each groups of DC was 94.2±5.2% (LMSN and AdmIL-12 cotransfected group), 78.9±3.6% (LMSN transfected groups), 52.6±9.7% (AdmIL-12 transfected group), 34.7±4.3% (LXSN transfected group), 36.3±3.8% (AdBGFP transfected group) and 3.9±2.0% (non-transfected group) respectively. Except for LXSN transfected and AdBGFP transfected group, the difference of the lytic activities between other groups were statistically significant (P<0.05). Conclusion: The MAGE-1 gene modified DCs can induce relatively specific cytotoxicty against SMMC7721 in vitro and thus suggested that those genetically engineered DCs have the potential to serve as novel vaccine for HCC. Transduction of exogenous IL-12 gene into DCs may further enhance DCs’ activity and the effectiveness of the above DC vaccine.展开更多
基金This work was supported by NationalNatural Science Foundation of China (No. 39770826) andpartly by CMB (No. 93583).
文摘Objective: to transduce the tumor associated antigen gene MAGE-1 and/or IL-12 gene into dendritic cells (DC) and to observe the in vitro cytotoxic effect induced by the genetically modified DC against the human hepatocellular carcinoma (HCC) cell line SMMC7721. Methods: the MAGE-1 gene was inserted into the retrovirus vector LXSN to construct the recombinant retrovirus LMSN. The monocyte-derived DCs were transfected at appropriate differentiation stage by LMSN and/or a recombinant adenovirus AdmiL-12, which containing murine IL-12 gene. The control groups included retrovirus LXSN transfected, adenovirus AdBGFP transfected and non-transfected DCs. The MAGE-1 gene expression was identified by western blot and the mIL-12 p70 secretion was detected by ELISA assay. The in vitro cytotoxicities against SMMC7721 induced by genetically modified and control groups of DC were tested by MTT assay. Results: The MAGE-1 expression was detected by a monoclonal antibody in DCs tranfected with LMSN but not in control groups. At 16 h, 24 h and 48 h after transfection with AdmIL-12, the concentration of the mIL-12 p70 in the culture medium was 580pg/106 cells, 960pg/106 cells and 1100pg/106 cells respectively. The mIL-12 p70 secretions were not detected in other groups. The lytic activity (as judged by % lysis) induced by each groups of DC was 94.2±5.2% (LMSN and AdmIL-12 cotransfected group), 78.9±3.6% (LMSN transfected groups), 52.6±9.7% (AdmIL-12 transfected group), 34.7±4.3% (LXSN transfected group), 36.3±3.8% (AdBGFP transfected group) and 3.9±2.0% (non-transfected group) respectively. Except for LXSN transfected and AdBGFP transfected group, the difference of the lytic activities between other groups were statistically significant (P<0.05). Conclusion: The MAGE-1 gene modified DCs can induce relatively specific cytotoxicty against SMMC7721 in vitro and thus suggested that those genetically engineered DCs have the potential to serve as novel vaccine for HCC. Transduction of exogenous IL-12 gene into DCs may further enhance DCs’ activity and the effectiveness of the above DC vaccine.
