ABSTRACT
Objective:To study the biological effects of TCR on hepatoma cells by transfecting V?7 into lymphocytes.Methods:TCR V?7 gene was amplified by RT PCR and cloned to expression vector pLXSN. The recombinant was transferred into lymphocytes by Lipofectin Reagent transfection, then the lymphocytes were co cultured with hepatoma cells. The phenotype of lymphocytes was detected on the Flow Cytometry, the expression of TCR V?7.1 gene was detected by Laser Scanning Confocal Microscope ( LSCM) and the ultrastructure of the hepatoma cells was showed by electronic microscope.Results:The amount of the transmembrane protein expressed by TCR V?7.1 gene was increased significantly, and so was the amount of lymphocytes (P
ABSTRACT
Objective:To investigate the immune state and the alteration of cytotoxicity to hepatocellular carcinoma cells BEL 7402 in hTCRV ? 8 4 gene transfeced PBMC after co culturing with HBV derived hepatocellular carcinoma cells BEL 7402. Methods:The hTCRV? 8 4 recombinant plasmid was transfered into peripheral blood mononuclear cells (PBMC).Flowcytometer analysis were used to assay the expression of TCRV?8.4 gene and the the immune state of hTCRV?8.4 gene modified PBMC.LDH release assay were used to test the cytotoxicity of the PBMC to hepatocellular carcinoma cells BEL 7402.Results:The expresstion of TCRV?8.4 in CD3 +T cell was increased significantly after gene transfection.The percentage of TCRV?8.4?CD122 + and CD19 +lymphocytes increased obviously after co cultured with BEL 7402,this indicates BEL 7402 stimulated the proliferation of these cells and cellular immunity.Transmission electronic microscope showed apoptosis in BEL 7402 induced by hTCRV?8.4 gene transferred PBMC.LDH release assay reveals increased cytotoxicity of hTCRV ?8.4 gene transfered PBMC to BEL 7402.Conclusion:The proliferation and the cytotoxicity were enhanced significantly in hTCRV ?8.4 modified PBMC after stimulated by HBV derived hepatocellular carcinoma cells BEL 7402.
ABSTRACT
Objective:To study the biological effects of TCR to hepatoma cell by transfection V?7 to lymphocytes. Methods:TCRV?7 gene was amplified by RT PCR and cloned to expression vector pLXSN. The recombinant was transferred into lymphocytes by Lipfectin Reagent transfection,then the lymphocytes were co cultured with hepatoma cells.The phenotype of lymphocytes was detected on the Flow Cytometry and the ultrastructure of the hepatoma cells was showed by electronic microscope.Results:The lymphocyte amount with TCRV?7 expressing in those being transfected was much more than those no transfection.Apoptosis appeared in the hepatoma cells.Conclusion:TCRV?7 subfamily can recognize hepatoma antigen and stimulate T cell.