树突状细胞dendriticcellsdc由美国学者steinman等于1973年.docVIP

小鼠骨髓来源未成熟树突状细胞蛋白表达的分析 易发平1,2，焦庆昉1,2，符少月1,2，卜友泉1,2，刘革力2，宋方洲1,2（重庆医科大学：1基础医学院生物化学与分子生物学教研室，2分子医学与肿瘤研究中心，重庆 400016） [摘要] 目的 利用双向电泳和质谱技术对小鼠骨髓来源未成熟树突状细胞表达的蛋白质进行分析，为进一步探讨和利用DC(dendritic cells , DC)的免疫机制提供实验依据。 IL-4和GM-CSF诱导未成熟DC的分化，提取细胞总蛋白，定量。双向凝胶电泳分离蛋白质组分，胶体银染显色，PDQuest进行图像分析后选取蛋白点，胶内酶解后MALDI-TOF MS进行肽指纹图谱鉴定，MS-Fit分析质谱数据。 细胞生长情况和表面标志物检测表明获得了高纯度的未成熟树突状细胞。图像分析结果显示，树突状细胞中检测到了660±10个蛋白点，主要分布在分子量28 kDa～100 KDa，等电点5.0～9.0之间。鉴定了87个蛋白点，其中，与未成熟DC免疫调节功能相关的蛋白质有40个，占45.98%；与大分子代谢相关的有37个，占42.53%。 成功分析了小鼠骨髓来源未成熟树突状细胞表达的蛋白质。nalysis of protein expression in mouse myeloid dendritic cells Yi Faping1,2，Jiao Qingfang1,2，Fu Shaoyue1,2，B Youquan1,2，Liu Geli2，Song Fangzhou1,2（(1Department of Biochemistry and Molecular Biology, 2Molecular Medicine and Cancer Research Center; Chongqing Medical University ,Chongqing 40016,China）[Abstract] Objective To afford experimental proof for application of dendritic cells (DC), the protein, expressed in immature DC derived from mouse myeloid, were separated by 2-dimention electrophoresis (2-DE), and identified by peptide mass fingerprinting method. Methods The differentiation of immature DC were induced by interleukin-4 (IL-4) and granulocyte-macrophage colony-stimulating factor (GM-CSF) . [基金项目] 国家自然科学基金，重庆市自然科学基金（CSTC，2008BB5225） [作者简介] 易发平，男，博士，副教授。研究方向：分子生物学，发表论文60余篇。电话：（023E-mail：fazicq@163.com [通信作者] 宋方洲，电话：（023E-mail：fzsongcq@163.com The proteins were extracted from the DC, separated by 2-DE, stained by improved silver method, analyzed by PDQuest software, and identified by peptide mass fingerprinting method and MS-Fit. Results The high pure immature DC were obtained through detecting Cell growth and surface markers. 660 ± 10 protein spots were successfully separated. Most of the molecular weight of protein were from 28 kDa to 100 Kda, and isoelectric point from 5.0 to 9.0. 87 spots were analyzed. A total of 40 proteins (45.98%) were related to immune functions and 37 proteins (42.53%) related to macromolecul