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


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.


CONCLUSIONS
The cytolytic T cell activity induced by the fusions of ovarian carcinoma cells to transfected DC is enhanced.
Peng
Ping
P
Department of Obstetrics and Gynecology, Peking Union Medical College Hospital, Peking Union Medical College, Chinese Academy of Medical Science, Beijing 100730, China.
Shen
Keng
K
He
Wei
W
Wu
Ming
M
Wei
Wei
W
Lang
Jing-he
JH
Zhang
Su-mei
SM
Qi
Jin
J
Hu
Yu
Y
Zhao
Jian-qing
JQ
D002459:Cell Fusion; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000166:cytology* / Q000276:immunology; D005260:Female; D005312:Fetal Blood / Q000166:cytology; D006801:Humans; D018664:Interleukin-12 / Q000235:genetics* / Q000378:metabolism; D008213:Lymphocyte Activation / Q000276:immunology; D010051:Ovarian Neoplasms / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D013601:T-Lymphocytes / Q000276:immunology; D014162:Transfection
D018664:Interleukin-12