title: Ovarian carcinoma cells influence differentiation of Lin-CD45RA- dendritic cell precursors into two mature subtypes in vitro.
abstract: OBJECTIVE
  Decreased number and impaired function of dendritic cells (DCs) have been found in ovarian carcinoma microenvironment. The study was designed to detect if this phenomenon was associated with abnormal DC differentiation influenced by ovarian carcinoma cells.
  
  
  METHODS
  FLT-3L and SCF were used for expanding DC precursors from CD34+ progenitors. GM-CSF and TNF-alpha were used to induce mature DCs. Supernatants of cultured ovarian carcinoma cell line SKOV3 were added, in order to study their influence on the differentiation and maturation of Lin-CD45RA- DC precursors. Flow cytometry was used to analyze cell subtypes and molecular surface markers. Allogeneic T-cell proliferation assay was used to exam stimulatory activity of DCs. IL-12 secretion was tested by ELISA.
  
  
  RESULTS
  Lin-CD45RA- DC precursors cultured with GM-CSF and TNF-alpha generated HLA-DR+CD11C+CD123- myeloid DCs (mDCs) and HLA-DR+CD11C-CD123+ plasmacytoid DCs (pDCs) in vitro. The supernatants from ovarian carcinoma cell line SKOV3 (SKOV3-supernatants) increased pDCs and decreased mDCs compared with pure medium or supernatants of normal ovarian surface epithelial (OSE) cells. There were no significantly different expressions of HLA-DR and CD80 by DCs between with and without SKOV3-supernatants. But DCs treated with SKOV3-supernatants were shown to have impaired immune activity to stimulate proliferation of allogeneic CD3+ T cells and secrete IL-12.
  
  
  CONCLUSIONS
  Ovarian carcinoma cells influence differentiation of Lin-CD45RA- DC precursors into subtypes of mature DCs in vitro. This resulted in fewer mDCs, increased number of pDCs, and impairment of mature DCs immune activity.
authors.affiliation: Women's Reproductive Health Laboratory of Zhejiang Province, Women's Hospital, School of Medicine, Zhejiang University, Hangzhou, China.
authors.forename: Li-li
authors.initials: LL
authors.lastname: Chen
authors.forename: Feng
authors.initials: F
authors.lastname: Ye
authors.forename: Yan
authors.initials: Y
authors.lastname: Yu
authors.forename: Huai-zheng
authors.initials: HZ
authors.lastname: Chen
authors.forename: Wu-wen
authors.initials: WW
authors.lastname: Zhang
authors.forename: Wei-guo
authors.initials: WG
authors.lastname: Lü
authors.forename: Xing
authors.initials: X
authors.lastname: Xie
mesh_terms: D000328:Adult; D002454:Cell Differentiation / Q000276:immunology*; D003713:Dendritic Cells / Q000276:immunology* / Q000473:pathology; D005260:Female; D005312:Fetal Blood / Q000276:immunology; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D006801:Humans; D017493:Leukocyte Common Antigens / Q000276:immunology*; D008213:Lymphocyte Activation; D008565:Membrane Proteins / Q000494:pharmacology; D010051:Ovarian Neoplasms / Q000276:immunology* / Q000473:pathology; D019089:Stem Cell Factor / Q000494:pharmacology; D013234:Stem Cells / Q000276:immunology / Q000473:pathology; D013601:T-Lymphocytes / Q000276:immunology; D018417:Th1 Cells / Q000276:immunology; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
chemical_list: D008565:Membrane Proteins; D019089:Stem Cell Factor; D014409:Tumor Necrosis Factor-alpha; C084881:flt3 ligand protein; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D017493:Leukocyte Common Antigens