title: [Role of glycogen synthase kinase 3β in maturation and function of murine myeloid dendritic cells in vitro].
abstract: OBJECTIVE
  To investigate the role of glycogen synthase kinase 3β (GSK-3β) in the maturation and function of murine bone marrow-derived dendritic cells (BMDCs).
  
  
  METHODS
  Mature DCs (mDCs) induced by LPS were examined for GSK-3β phosphorylation level with Western blotting before and after LPS exposure. To explore the role of GSK-3β in maturation and function of DCs, we added SB216763, a selective inhibitor of GSK-3β, in the cell culture of immature DCs (iDCs), and examined CD40 and CD86 expressions in the cells by flow cytometry and the expression of IL-6, IL-12 and IL-10 mRNA by real-time PCR; the changes of the immunogenicity of the cells was evaluated by mixed lymphocyte reaction. The expression of GSK-3β and RelB was examined by Western blotting in DC2.4 cells transfected with a lentiviral vector over-expressing murine GSK-3β gene.
  
  
  RESULTS
  LPS exposure significantly lowered GSK-3β activity in iDCs as demonstrated by increased Ser9 phosphorylation and reduced Tyr216 phosphorylation. GSK-3β inhibition induced DC maturation by increasing the expression of surface costimulatory molecules CD40 and CD86, lowered the expressions of IL-6 and IL-12 while enhanced the expression of IL-10 in iDCs, and impaired mixed lymphocyte reaction of the cells. In DC2.4 cells, lentivirus-mediated over-expression of GSK-3β obviously down-regulated the expression of RelB.
  
  
  CONCLUSIONS
  GSK-3β is a crucial enzyme involved in the differentiation and maintenance of an immature phenotype of DCs. GSK-3β is constitutively active in iDCs to inhibit their spontaneous maturation. DCs become phenotypically mature after inhibition of GSK-3β, which also executes a proinflammatory task in DC activation. The reduction of RelB protein levels as a result of GSK-3β overexpression supports GSK-3β as a new target for inducing tolerogenic DCs.
authors.affiliation: Laboratory Medicine Center, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China. E-mail: nh.cs@163.com.
authors.forename: Shuai
authors.initials: S
authors.lastname: Chu
authors.forename: Haixia
authors.initials: H
authors.lastname: Li
authors.forename: Xin
authors.initials: X
authors.lastname: Li
authors.forename: Xia
authors.initials: X
authors.lastname: Kang
authors.forename: Qingshui
authors.initials: Q
authors.lastname: Huang
authors.forename: Hongxia
authors.initials: H
authors.lastname: Wang
authors.forename: Yurong
authors.initials: Y
authors.lastname: Qiu
mesh_terms: D000818:Animals; D051940:B7-2 Antigen / Q000378:metabolism; D019013:CD40 Antigens / Q000378:metabolism; D002454:Cell Differentiation; D002478:Cells, Cultured; D003470:Culture Media / Q000737:chemistry; D003713:Dendritic Cells / Q000201:enzymology*; D038362:Glycogen Synthase Kinase 3 / Q000378:metabolism*; D000071679:Glycogen Synthase Kinase 3 beta; D007211:Indoles / Q000737:chemistry; D016753:Interleukin-10 / Q000378:metabolism; D018664:Interleukin-12 / Q000378:metabolism; D015850:Interleukin-6 / Q000378:metabolism; D016086:Lentivirus; D007959:Lymphocyte Culture Test, Mixed; D008301:Maleimides / Q000737:chemistry; D051379:Mice; D022423:Myeloid Cells / Q000201:enzymology*; D010766:Phosphorylation; D012333:RNA, Messenger; D060888:Real-Time Polymerase Chain Reaction; D015398:Signal Transduction
chemical_list: D051940:B7-2 Antigen; D019013:CD40 Antigens; D003470:Culture Media; D007211:Indoles; D015850:Interleukin-6; D008301:Maleimides; D012333:RNA, Messenger; C417521:SB 216763; D016753:Interleukin-10; D018664:Interleukin-12; D000071679:Glycogen Synthase Kinase 3 beta; C000605507:Gsk3b protein, mouse; D038362:Glycogen Synthase Kinase 3