PMID: 16635331Apr 26, 2006Paper

Primary study on fusions of ovarian carcinoma cells to dendritic cell transfected with interleukin-12 gene in vitro

Zhonghua fu chan ke za zhi
Ping PengJian-qing Zhao

Abstract

To investigate the immune response of the fusions of ovarian carcinoma cells to dendritic cell (DC) transfected with interleukin (IL)-12 gene in vitro. Recombinant human granulocyte macrophage colony stimulating factor and recombinant human tumor necrosis factor alpha were used to generate DC from cord blood in vitro. IL-12 fusion gene was cloned into a eukaryotic vector-pcDNA3.1 (+). DC were transfected with IL-12-pcDNA3.1 (+) using lipofectamine transfection method and electric transfection method respectively. Then polyethylene glycol mediated the fusion of transfected DC and ovarian carcinoma cells, and 3-(4, 5-dimethylthiazol-2-yl) 2, 5-diphenyl tetrazolium bromide (MTT) test was carried out to observe the immune response. DC were generated in vitro from cord blood and expressed high levels of costimulatory (50%) and MHC II molecules (99%). RT-PCR showed that IL-12-pcDNA3.1 (+) had been successfully transfected into DC. RT-PCR and immunofluorescence analysis showed that DC were fused with ovarian carcinoma cells successfully. Then the fusion cells of ovarian carcinoma cells to transfected DC, and the fusions of ovarian carcinoma cells to DC were cocultured with peripheral blood mononuclear cell respectively. MTT test showe...Continue Reading

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