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Effect of monoclonal antibody to CD47 molecule on dendritic cell differentiation and function
Authors:YU Jing  LIN Mao-fang  MOU Hai-bo  HUANG He  CAI Zhen
Affiliation:Department of Hematology,The First Affiliated Hospital, Medical College of Zhejiang University, Hangzhou 310003, China
Abstract:AIM: To explore the influence of CD47 molecules on the maturation and function of cultured dendritic cells (DCs). METHODS: Monocyte cell-derived DCs were propagated in granulocyte-macrophage colony-stimulating factor (GM-CSF) and lipopolysaccharide (LPS) plus interleukin (IL)-4, in the presence or absence of anti CD47 monoclonal antibodies (anti-CD47 mAbs). Flow cytometry was used to detect the cell surface phenotype. The concentration of IL-12P70 in supernatant was measured by ELISA technique. The antigen-presenting functions of DCs were determined in one-way mixed leukocyte reaction by Brdu-ELISA. Electrophoretic mobility shift assays (EMSA) was used to examine NF-κB activity. RESULTS: The anti-CD47 mAbs markedly suppressed the expression of CD80,CD86,CD83,CD1a,HLA-DR on the surface of DCs (P<0.05). The data of mixed leukocyte reaction and IL-12P70 production were consistent with the flow cytometry results (P<0.01). Nuclear extracts from the anti-CD47 mAb-treated DCs revealed a decrease in NF-κB binding activity. CONCLUSION: The anti-CD47 mAb exerts a negative effect on the maturation and function of in vitro cultured DCs via inhibiting of NF-κB binding activity.
Keywords:Dendritic cells  Immune tolerance  NF-kappa B   
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