摘要
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。
We aimed to investigate the effects of Fucoidan,a non-lipoprotein ligand for SR-A,on the monocyte derived DC（MDDC） in vitro.We isolate monocyte and induced the cells differentiate into MDDC by treating with GM-CSF plus IL-4.Then we collected the MDDC and treated them with Fucoidan or LPS for 24 hours.Subsequently we detected the expression of cell surface marker and phagocytic capacity by flow cytometry.ELISA was used to detect the levels of cytokine secretion by MDDC and Th cells;T cell proliferation was assessed by CFSE staining.The results showed that in the presence of Fucoidan,the surface markers on MDDC,such as CD80,CD83 and MHC-Ⅱ,were up-regulated,among which the up-regulation of CD83 followed the dose-dependant manner.Fucoidan also decreased the phagocytic capacity of mature MDDC（Fucoidan-mDC）,but stimulated MDDC to produce IL-10,IL-12p40 and TNF-α.In conclusion,Fucoidan can induce considerable Th cell activation and proliferation,which would elucidate the function of Fucoidan and SR-A in activation and maturation of DC.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2010年第11期934-937,共4页
Immunological Journal
基金
国家自然科学基金(30960358
30860263)
