title: [Echinococcus granulosus recombinant antigen B induced IDO expression in mouse bone marrow-derived dendritic cells in vitro].
abstract: OBJECTIVE
  To observe the expression of indoleamine 2,3-dioxygenase (IDO) in mouse bone marrow-derived dendritic cells (DCs) after adding Echinococcus granulosus recombinant antigen B (rAgB) in vitro.
  
  
  METHODS
  CD11c+ DCs generated from bone marrow precursor cells of C57BL/6 mice and cultured in the presence of recombinant mouse GM-CSF (rmGM-CSF). The morphology of DCs was observed by inverted microscope and scanning electronic microscope. The level of I-A/I-E, CD40, CD80, and CD86 on DCs were determined by flow cytometry. T cell proliferation induced by DCs were evaluated by using mixed lymphocyte reaction (MLR) assay. At day 6 post culture, the immature DCs were collected, and part of the immature DCs stimulated with lipopolysaccharide (LPS) for 24 h were examined by flow cytometry. Immature DCs were divided into 3 groups: negative control group, positive control group (rmIFN-gamma, 1000 U/ml) and rAgB group. Immature DCs of positive control group and rAgB group were induced with 1000 U/ml rmIFN-gamma and 15 microg/ml rAgB, respectively. IDO expression in DCs was examined 24 h after induction using immunohistochemical method and Western blotting.
  
  
  RESULTS
  More than 80% CD11c+ DCs were harvested. The typical DCs were observed under inverted microscope and scanning electronic microscope. The level of CD40, CD80, and IA/IE (MHC II) in mature DCs group was significantly higher than that of immature DCs group (P < 0.05). In MLR, mitomycin-treated DCs can stimulate T lymphocytes proliferation activity. There were significantly differences in IDO expression in the negative control group [(4.544 +/- 1.752)%], positive control group [(20.464 +/- 4.452)%] and rAgB group [(11.148 +/- 1.966)%] (P < 0.05). Western blotting result indicated that the ratio of IDO/GAPDH in rAgB group (0.573 +/- 0.129) was significantly higher than that of negative group (0.229 +/- 0.085) (P < 0.05), and there were no significant difference in the ratio of IDO/GAPDH between IFN-gamma group (0.794 +/- 0.114) and rAgB group (P > 0.05).
  
  
  CONCLUSIONS
  rAgB can induce IDO expression in bone marrow-derived dendritic cells in vitro.
authors.affiliation: Xinjiang Laboratory of Hydatid Fundamental Medicine, the First Affiliated Hospital, China.
authors.forename: Jiao-yu
authors.initials: JY
authors.lastname: Shan
authors.forename: Wei-zheng
authors.initials: WZ
authors.lastname: Ji
authors.forename: Turgun
authors.initials: T
authors.lastname: Tursun
authors.forename: Liang
authors.initials: L
authors.lastname: Li
authors.forename: Chuan-shan
authors.initials: CS
authors.lastname: Zhang
authors.forename: Ren-yong
authors.initials: RY
authors.lastname: Lin
authors.forename: Hao
authors.initials: H
authors.lastname: Wen
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000166:cytology / Q000276:immunology / Q000378:metabolism*; D050503:Indoleamine-Pyrrole 2,3,-Dioxygenase / Q000378:metabolism*; D007371:Interferon-gamma / Q000276:immunology; D008074:Lipoproteins / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D013601:T-Lymphocytes / Q000276:immunology
chemical_list: D050503:Indoleamine-Pyrrole 2,3,-Dioxygenase; D008074:Lipoproteins; C514022:antigen B, Echinococcus granulosus; D007371:Interferon-gamma