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目的探讨清道夫受体A(SR-A)非脂蛋白配体,褐藻多糖硫酸酯(Fucoidan),对人体外培养的单核诱导的树突状细胞(monocyte derived DC,MDDC)的影响及其机制。方法采用免疫磁珠法,直接分离外周血单核细胞并通过GM-CSF,IL-4诱导生成MDDC,经Fucoidan或LPS刺激后,用流式细胞技术(FCM)检测树突状细胞表面协同刺激分子表达及细胞吞噬能力的变化;酶联免疫吸附法(ELISA)检测MDDC及Th细胞分泌的细胞因子;CFSE法检测经刺激后MDDC对Th细胞活化、增殖的影响。结果经流式细胞分析术检测,Fucoidan刺激后的MDDC膜表面CD80、CD83及MHC-Ⅱ分子的表达均增强,其中CD83分子表达上调呈剂量依赖性,同时MDDC吞噬能力减弱;IL-10,IL-12p40和TNF-α的分泌增加;并且可以诱导Th细胞的分泌IFN-γ并诱导其增殖。结论 Fucoidan在体外可以诱导MDDC的成熟、活化,刺激其分泌相关细胞因子,发挥其免疫应答中抗原呈递,促进Th活化、增殖分化的作用;通过该作用阐明Fucoidan及SR-A在DC的活化和成熟过程中的作用。
Objective To investigate the effects of scavenger receptor A (SR-A) non-lipoprotein ligand and fucoidan on monocyte derived DC (MDDC) cultured in vitro mechanism. Methods Immunomagnetic beads method was used to directly separate peripheral blood mononuclear cells and induce MDDC through GM-CSF and IL-4. After stimulation with Fucoidan or LPS, flow cytometry (FCM) was used to detect the synergistic stimuli of dendritic cells The expression of cytokines and the changes of phagocytosis were detected by enzyme-linked immunosorbent assay (ELISA). The cytokines secreted by MDDC and Th cells were detected by ELISA. The effects of MDDC on the activation and proliferation of Th cells were detected by CFSE. Results The expression of CD80, CD83 and MHC-Ⅱ on the surface of MDDC membrane stimulated by Fucoidan were all increased by flow cytometry. The up-regulation of CD83 expression was dose-dependent and phagocytosis of MDDC was diminished. The levels of IL-10, IL -12p40 and TNF-α secretion; and can induce the secretion of IFN-γ Th cells and induce their proliferation. Conclusion Fucoidan can induce the maturation and activation of MDDC in vitro and stimulate the secretion of related cytokines and exert the antigen presentation in immune response and promote the activation of Th and the proliferation and differentiation of MDDC. The effects of Fucoidan and SR-A on DC activation and The role of maturation process.