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一株特殊功能的抗人CD40单克隆抗体的获得及其生物学特性分析 |
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周照华 王江方 王月丹 邱玉华 潘建忠 谢玮 蒋令瑜 张学光
摘 要 目的：研制功能性抗人CD40单克隆抗体，从而探讨其对B细胞、DCs表达的CD40分子激发所产生的生物学效应。方法：采用细胞融合、单克隆抗体筛选、荧光标记、免疫印迹和竞争抑制等方法获得鼠抗人CD40mAb，以细胞增殖、分化抗原表达观察CD40mAb对B细胞和DCs的作用效应。结果：经表型分析、Western blotting鉴定和竞争抑制试验，证实5C11是识别人CD40分子的特异性单抗；5C11与转导CD32的L细胞（LCD32）联合IL-4能使扁桃体B细胞扩增并形成集落；5C11能介导树突状细胞（DCs）的扩增和分化成熟。结论：用5C11与LCD32建立的CD40培养体系，能使B细胞长期生存和增殖，为体外研究B细胞提供有效手段；5C11诱导外周单核细胞分化成熟为树突状细胞，这一发现为体外大规模扩增可用于治疗的DCs提供了新的手段，因而5C11是一株具有特殊功能和重要应用价值的单克隆抗体。
关键词 CD40 单克隆抗体 B细胞 树突状细胞 多发性骨髓瘤
中国图书分类号 R392.11
The obtaining of an anti-human CD40 mono-clonal antibody with special functions and the analysis of it s biological effects
ZHOU Zhao-Hua, WANG Jiang-Fang, WANG Yue-Dan et al.
Department of Immunology, Suzhou Medical College, Suzhou 215007
Abstract Objective： To prepare mouse anti-human CD40 antigen functional monoclonal antibody and to further study it s biological effects by triggering the CD40 molecules on B cells and dendritic cells(DCs) . Methods: Using cell fusion, McAb screening , immunofluorescence analysis, immunoblotting and competition test to obtain mouse anti-human CD40 McAb； by the proliferation assay of B cells and DCs, and the analysis of expression of differentiation antigens on DCs. Results: On the basis of phenotype analysis, Western blotting and competition test, it is verified that 5C11 recognizes human CD40 antigens specially; 5C11 can augement the proliferation of tonsil B cells in the LCD32 cells and IL-4 culturing system; 5C11 can medicate DCs to get proliferation and maturation.Conclusion: 5C11+LCD32+IL-4 can make tonsil resting B cells survive and long-term proliferate in vitro, the setup of CD40 system furnish necessary tool for the study of B cells; McAb 5C11 also triggered the generation, proliferation and maturation of dendritic cells from peripheral blood monocytes. Thus 5C11 is a McAb with special function and important application value.
Key words CD40 McAb B cells Den[1] [2] 下一页
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