Résumé
Objective:To observe lethal effect on liver cancer cells by the DCs transfected with recombinant plasmid pchIL -18-MAGE.Methods:Cultivate dendritic cells in vitro ,and recombinant plasmid pchIL-18-MAGE was constructed which was transfected into DCs.Expression of IL-18 and MAGE-1 on the transfected DCs were detected by RT-PCR and Western blot.The diversity of DCs phenotypes after transfect were analyzed by flow cytometry.Different killing effects to the target cells by lymphocytes which were stimulated by DC and those by non-transfected DC were detected.IFN-γin the cellular supernatant of the transfected DCs was inspected by ELISA.Results:pchIL-18-MAGE transfected DCs expressed higher level of CD 83,CD1a,CD86,CD80,HLA-DR,and expressed as a mature DC phenotype.pchIL-18-MAGE group′s killing activity was the highest ( P<0.05 ) .Conclusion: The DC vaccine pchlL-18-MAGE can induce better killing effect compared with other experimental groups.
Résumé
Objective:To observate immunologic response of immunized mice by the vaccine pcIL-18-MAGE.Methods:Abundant plasmids were extracted and immunized mice,two booster injection were carried out at ten days intervals.Seven days after the third immunization,serum antibody against MAGE-1 and T cell subgroup were detected by FCM,and CTL killing activity were assayed through MTT.Results:About serum antibody against MAGE-1 and T cell subgroup,recombinant plasmids groups were all higher than the vacant plasmid pcDNA3 group(P
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Objective:To construct pEGFP-C3-B7.2-MAGE-1 eukaryotic expression plasmids and to study the immunological effects of esophageal cancer cell vaccine modified by human B7.2/MAGE-1 gene.Methods:The B7.2 and MAGE-1 cDNA,which was amplified by RT-PCR, and the eukaryotic expression plasmid pEGFP-C3-B7.2-MAGE-1 was constructed. Human esophageal cancer cell EC9706 was transfected with the vector of pEGFP-C3-B7.2-MAGE-1 using the technique of lipofectamine transfection.The dendritic cells(DCs)from peripheral blood mononuclear cells(PBMC)were loaded with the tumor antigen,and co-cultured with congeneric T cells derived from PBMCs for 3 days to obtain the tumor specific cytotoxicity T lymphocytes(CTL). Methy1 thiazoly1 tetrazolium (MTT) assay was used to detect inhibition effect of CTL on transfected and untransfected EC9706 cells.Results:The CTL had stronger inhibition response against the cancer cells transfected with pEGFP-C3-B7.2-MAGE-1 than to the cancer cells transfected with pEGFP-C3 and the untransfected cancer cells(P
Résumé
Objective: To study the methylation state within MAGE 1 B′B promoter in gastric carcinoma and the association between demethylation and pathological differentiation, the association between demethylation and clinical stage. Methods: Using methylation sensitive restriction analysis followed by polymerase chain reaction (PCR),we studied 80 specimen that were obtained from surgical samples (including 40 gastric carcinoma and 40 matched adjacent normal gastric mucosae).Results: An demethylation state was identified in DNA from gastric carcinoma specimens.The demethvlation rate is 25%(10/40).In contrast,no demethylation state was identified in DNA from matched adjacent normal gastric mucosae. The differences were Significant statistically. ln our study, the demethylation in poorly, moderately, and well differentiated glandulous cell carcinoma were detected at frequencies of 50%,18.7% and 8.3%,respectively, The differences were significant statistically ( P