抗P选择素功能域单抗对人树突状细胞表型及IL-12分泌的影响

Influence of the Monoclonal Antibody Against Lectin-EGF Domain of P-Selectin Upon the Phenotype and Proinflammatory Cytokine Secretion of Cultured Human Dendritic Cells

观察抗P选择素Lectin EGF功能域单抗 (PsL EGFmAb )对体外培养人树突状细胞 (DC )表型以及促炎细胞因子IL 1 2分泌的影响 ,探讨PsL EGFmAb对DC炎性成熟过程中的调节作用。通过SCF、GM CSF、TGF β1 、Flt 3和TNF α体外培养体系 ,从脐血CD34+ 造血干细胞中诱导扩增获得DC ,并于成熟中用PsL EGFmAb进行干预。采用流式细胞仪分析细胞表型CD1a、CD1 1c、CD83、CD80、CD86和HLA DR ;采用RT PCR检测IL 1 2p35、p4 0mRNA表达 ;以及ELISA法测定IL 1 2p70分泌的含量。结果显示 ,PsL EGFmAb可下调成熟中DC表面CD1 1c、CD83、CD80、CD86和HLA DR的表达 ,同时能抑制DC内IL 1 2p35、p4 0mRNA的转录和IL 1 2p70的分泌。本研究提示 ,PsL EGFmAb对DC黏附共刺激分子表达和促炎细胞因子合成具有抑制作用 。

To observe the influence of the monoclonal antibody against Lectin-EGF domain of P-selectin (PsL-EGFmAb) upon the morphologic features of cultured human dendritic cells (DC), as well as the expression of IL-12p35,p40 mRNA and secretion of IL-12p70, CD34+ stem cells from cord blood were isolated and cultured in 20%IMDM medium with SCF,GM-CSF,TGF-β_1,Flt-3,TNF-α. DC were interfered by PsL-EGFmAb.CD1a,CD11c,CD83,CD80,CD86 and HLA-DR was assayed by flow cytometry. IL-12p35,40 mRNA was detected by RT-PCR、IL-12p70 secreted in culture medium was detected by ELISA. It was found that the expression of CD11c,CD83,CD80,CD86 and HLA-DR was lower in PsL-EGFmAb group, and the expression of IL-12p35,40 mRNA and the secretion of IL-12p70 was suppressed after PsL-EGFmAb treatmeut. The PsL-EGFmAb might inhibit the expression of adhesion and co-stimulation molecules, and inhibit proinflammatory cytokine secretion, as well as adjust the antigen presenting function of DC.