【摘要】 目的探讨甘露聚糖结合凝集素对细菌脂多糖诱导的人外周血单核细胞来源树突状细胞(MoDC)成熟的影响。方法从健康成人外周血分离能黏附塑料的单核细胞,在rhGMCSF和rhIL4条件下培养5d,然后在有或无LPS和不同质量浓度(10～100mgL)天然人MBL条件下继续培养2d。用倒置显微镜观察DC的形态,以FACS分析DC的表型,用MTT法测定DC刺激同种异体T细胞增殖的能力,以酵母多糖颗粒吞噬试验评估DC的抗原摄取能力,用ELISA检测DC培养上清液中TNFα和IL12p40+p70的含量。结果MBL以剂量依赖方式下调LPS诱导人MoDC表面分子CD83和CD86表达,增强其摄取酵母多糖颗粒的能力,降低其激发初始T细胞增殖的能力,并抑制其LPS诱导的TNFα和IL12p40+p70分泌,但MBL质量浓度低至10mgL时对LPS诱导DC成熟作用几无影响。结论高质量浓度MBL能抑制LPS诱导的DC成熟过程,提示其可能在LPS引发疾病包括败血症或感染性休克中具有调节作用,亦提供了分析DC分化成熟有关信号途径的新手段。更多还原

【Abstract】 Objective To investigate the effects of mannan-binding lectin (MBL) on the lipopolysaccharide (LPS) -induced maturation of human peripheral blood monocyte-derived dendritic cells (MoDCs). Methods The plastic-adherent monocytes were prepared from the blood of healthy adult volunteers. The MoDCs were induced by 5-day-culture in medium supplemented with rhGM-CSF and rhIL-4, and then cultured for 2 days in the presence or absence of LPS and human natural MBL at varying concentrations ranging from 10 to 100 mg/L. The morphology of DC was observed under inverted microscope and the expressions of CD83 and CD86 on DCs analyzed by FACS. The abilities of DCs to stimulate the proliferation of allogenic T cells and to up-take antigen were evaluated by MTT assay and zymosan granule phagocytosis test, respectively. The levels of TNF-α and IL-12 p40+p70 in culture supernatant of DCs were detected by ELISA. Results MBL down-regulated the LPS-induced expression of CD83 and CD86 on MoDCs, enhanced the ability of DCs to up-take zymozan granules, decreased the DC-indnced proli- feration of naive T cells, and inhibited the production of TNF-α and IL-12 p40+p70 by LPS in a dose-dependent manner. MBL, at lower concentration (10 mg/L), had no notable effects on LPS-induced maturation of DCs. Conclusion MBL can inhibit LPS-induced maturation of DCs, suggesting that MBL might play a role in controlling LPS-related disorders including sepsis and septic shock, which provides a new tool to dissect the signaling pathways involved in DC maturation.更多还原