Role of glycogen synthase kinase 3β in maturation and function of murine myeloid dendritic cells in vitro
10.3969/j.issn.1673-4254.2015.12.29
- VernacularTitle:GSK-3β对小鼠骨髓树突状细胞成熟和功能的调控作用
- Author:
Shuai CHU
1
;
Haixia LI
;
Xin LI
;
Xia KANG
;
Qingshui HUANG
;
Hongxia WANG
;
Yurong QIU
Author Information
1. 南方医科大学南方医院检验医学科
- Keywords:
GSK-3β;
dendritic cells;
maturation and function;
RelB
- From:
Journal of Southern Medical University
2015;(12):1809-1814
- CountryChina
- Language:Chinese
-
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-3βphosphorylation level with Western blotting before and after LPS exposure. To explore the role of GSK-3βin 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-3βand RelB was examined by Western blotting in DC2.4 cells transfected with a lentiviral vector over-expressing murine GSK-3βgene. Results LPS exposure significantly lowered GSK-3βactivity 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-3βobviously down-regulated the expression of RelB. Conclusion GSK-3βis 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.
