Primary study on fusions of ovarian carcinoma cells to dendritic cell transfected with interleukin-12 gene in vitro
- VernacularTitle:白细胞介素12转染树突状细胞后与新建卵巢上皮性癌细胞融合的免疫原性研究
- Author:
Ping PENG
;
Keng SHEN
;
Wei HE
;
Ming WU
;
Wei WEI
;
Jinghe LANG
;
Sumei ZHANG
;
Jin QI
;
Yu HU
;
Jianqing ZHAO
- Publication Type:Journal Article
- Keywords:
Dendritic cells;
Interleukin-12;
Ovarian neoplasms;
Transfection;
Cell fusion
- From:
Chinese Journal of Obstetrics and Gynecology
2001;0(01):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the immune response of the fusions of ovarian carcinoma cells to dendritic cell(DC) transfected with interleukin(IL)-12 gene in vitro. Methods Recombinant human granulocyte macrophage colony stimulating factor and recombinant human tumor necrosis factor ? 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. Results 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 showed that both fusions could induce cytolytic T cell activity and lysis of ovarian carcinoma cells,and the former effect was stronger. Conclusion The cytolytic T cell activity induced by the fusions of ovarian carcinoma cells to transfected DC is enhanced.
