GL50分子在单核细胞来源树突状细胞上的表达及其逆向信号对树突状细胞体外分化成熟的促进作用  

The Expression Analysis of GL50 on Monocyte-derived DCs(Mo-DCs) and Promotion of Maturation of Mo-DCs Stimulated by the Reverse Signaling of GL50

作　　者：王凤鸣[1] 刘彤[1] 孙静[1] 胡益洲[1] 王旭东[1] 张学光[1] 

机构地区：[1]苏州大学生物技术研究所,江苏苏州215007

出　　处：《苏州大学学报（医学版）》2007年第4期505-509,共5页Suzhou University Journal of Medical Science

基　　金：国家自然科学基金资助项目(No.30471609);江苏省高校自然科学研究计划项目(No.04KJB310124)

摘　　要：目的分析GL50在不同分化时期单核细胞来源树突状细胞(Mo-DCs)上的表达,探讨GL50信号对Mo-DCs的生物学作用。方法用Ficoll密度离心及贴壁法获得外周血单核细胞,经GM-CSF+IL-4常规方法诱导,并分别加入丝裂霉素处理的mock-L929和ICOS-L929细胞,6 d时,加入或不加入TNF-α培养3 d后收集细胞;另外,在上述ICOS-L929细胞处理组的单核细胞中加入不同浓度的GL50单克隆抗体11C4或小鼠同型IgG,加入或不加入TNF-α,继续培养3 d后,收集细胞,免疫荧光标记和流式细胞术分析Mo-DCs表型(GL50、CD80、CD83、CD86)、摄取FITC-Dextran抗原的能力;混合淋巴细胞反应检测Mo-DCs对T细胞的作用;ELISA方法检测对分泌细胞因子IL-2I、L-10的影响。结果GL50分子在不成熟Mo-DCs上高表达,而随着Mo-DCs的成熟呈下调性表达;GL50单克隆抗体11C4能够有效地促进Mo-DCs的成熟,上调Mo-DCs表面分子(CD86、CD80、CD83、HLA-DR)的表达,降低Mo-DCs摄取FITC-Dextran的能力,同时能有效地促进细胞因子IL-10的分泌,而对IL-2的分泌没有显著影响。结论GL50分子介导的逆向信号参与了Mo-DCs分化成熟的调节,GL50/ICOS是参与DC/T细胞相互作用的主要分子。Objective To detect the GL50 expression on Mo-DCs and analyze the role of GL50 in regulation of Mo-DCs development and functions. Methods Peripheral blood mononuclear cells （PBMC） heparinized blood were isolated by Ficoll density gradient centrifugation, and PBMCs were plated into 6-weU plates for 3 hours incubation. The non-adherent cells were removed and the adherent cells （monocytes） were cultured with GM-CSF and IL-4 with ICOS-L929 or mock-L929 cells for 5 days. After Mo-DCs were stimulated by anti-CD40 monoclonal antibody or TNF-α with or not stimulated with monoclonal antibody 11C4, the cells were harvested on day 2 and reacted with PE-conjugated CD50, CD86, and CD83. Then, cell surface molecules including GL50, CD50, CD83 and CD86 were detected by flow cytometry. The induced DC functions were estimated by detecting the capacity of DC uptake antigens with FITC-Dextran capturing assay. Interleukin-2 （IL-2） and IL-10 were measured with the help of ELISA kits. Results We found that the expression of GL50 molecule was only detected on the surface of immature Mo-DCs, and was sharply decreased after induction of mature Mo- DCs activated by TNF-α or CD40. Furthermore, we demonstrated that reverse signal of GL50 could effectively promote the maturation of Mo-DCs in vitro as evidenced by the high expression of CD80, CD86 and CD83 as well as by with the low capacity of uptaking FITC-Dextran particle, which suggested that GL50 may be involved in the maturation of Mo-DCs. Conclusion These results suggest that reverse signal of GL50 could restrain the differentiation and maturation of monocyte-derived DC.

关 键 词：GL50 树突状细胞 逆向信号 混合淋巴细胞反应 细胞因子 

分 类 号：R392.11[医药卫生—免疫学]