【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、Ｗｅｓｔｅｒｎｂｌｏｔｔｉｎｇ鉴定和竟争抑制试验，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。更多还原

【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 obtain mouse anti-human CD40 McAb;by the proliferation assay of B cells and DCsand 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 5Cll recognizes human CD40 antigens specially; 5Cll can augement the proliferation of tonsil B cells in the LCD32cells and IL-4 culturing system; 5Cll can medicate DCs to get proliferation and maturatio. Conclusion: 5Cll+LCD32 +IL4 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; McAb5Cll also triggered the generation, proliferation and maturation of dendritic cells from peripheral blood monocytes. Thus 5Cll is a McAb withspecial function and important application value.更多还原