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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。
Objective: To develop a functional anti-human CD40 monoclonal antibody to explore its biological effects on CD40 molecules expressed by B cells and DCs. Methods: Mouse anti-human CD40 mAb was obtained by cell fusion, monoclonal antibody screening, fluorescent labeling, western blot and competition inhibition. The effect of CD40 mAb on B cells and DCs was observed by cell proliferation and differentiation antigen expression. Results: Phenotypic analysis, Western blotting and competitive inhibition assays confirmed that 5C11 was a specific monoclonal antibody that recognizes human CD40 molecule. 5C11 combined with L-cell transduction of CD32 (LCD32) and IL-4 could amplify tonsil B cells And formed colonies; 5C11 can mediate dendritic cells (DCs) expansion and differentiation and maturation. Conclusion: The CD40 culture system established by 5C11 and LCD32 can make B cells survive and proliferate for a long time, and provide an effective method for studying B cells in vitro. 5C11 induces differentiation and maturation of peripheral mononuclear cells into dendritic cells. 5C11 is a monoclonal antibody that has special function and important application value because of scale-up. It provides a new method for the treatment of DCs.