title: [Primary study on fusions of ovarian carcinoma cells to dendritic cell transfected with interleukin-12 gene in vitro].
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 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.
authors.affiliation: Department of Obstetrics and Gynecology, Peking Union Medical College Hospital, Peking Union Medical College, Chinese Academy of Medical Science, Beijing 100730, China.
authors.forename: Ping
authors.initials: P
authors.lastname: Peng
authors.forename: Keng
authors.initials: K
authors.lastname: Shen
authors.forename: Wei
authors.initials: W
authors.lastname: He
authors.forename: Ming
authors.initials: M
authors.lastname: Wu
authors.forename: Wei
authors.initials: W
authors.lastname: Wei
authors.forename: Jing-he
authors.initials: JH
authors.lastname: Lang
authors.forename: Su-mei
authors.initials: SM
authors.lastname: Zhang
authors.forename: Jin
authors.initials: J
authors.lastname: Qi
authors.forename: Yu
authors.initials: Y
authors.lastname: Hu
authors.forename: Jian-qing
authors.initials: JQ
authors.lastname: Zhao
mesh_terms: 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
chemical_list: D018664:Interleukin-12