中文摘要：

CD40是肿瘤坏死因子受体超家族成员之一,属于Ⅰ型跨膜糖蛋白,其自然配体是CD40L。CD40-CD40L信号通路在许多重要的生理和病理过程中都起到关键的调节作用。基于其在激活免疫应答中的重要作用,在肿瘤免疫治疗中,具有激活CD40-CD40L信号的CD40激动剂抗体可用于提高抗肿瘤免疫应答;在移植排斥和炎症反应中,CD40-CD40L信号也起重要作用,阻断CD40信号通路是目前治疗风湿性关节炎、肠炎、系统性红斑狼疮、多发性硬化和糖尿病的一个重要靶点。制备CD40阻断性抗体或激动剂抗体不仅可用于检测CD40分子表达,还可以开发用于炎症性疾病或肿瘤治疗的免疫疗法。本研究表达CD40胞外段蛋白,通过免疫制备14株CD40单克隆抗体,筛选发现全部可作为western blotting检测CD40蛋白表达的抗体,9株CD40抗体可作为一抗用于流式细胞术检测人CD40表达,1株CD40抗体能阻断人CD40L和CD40相互作用。有意义的是,8株CD40抗体能激活人CD40分子信号通路,另外有10株能激活小鼠脾脏细胞CD40下游基因的表达,7株能上调小鼠脾脏B细胞表面共刺激分子CD86的表达。本研究制备一系列CD40抗体,可用于CD40检测、CD40信号的阻断或激活。

英文摘要：

The preparation of blocking or antagonistic antibodies to CD40 can not only be used to detect the expression of CD40,but also to the treatment of inflammatory diseases or tumor immunotherapy.By using a recombinant CD40 protein,we generated fourteen hybridoma cell lines to produce CD40 antibodies.All the CD40 monoclonal antibodies could be used to detect the CD40 expression via western blotting,nine of which can also be used for FACS and one CD40 monoclonal antibody can block the CD40-CD40 Linteraction.Importantly,we got eight clones that produced agonistic antibodies to human CD40,while ten of the clones could activate the CD40 downstream gene expression of mouse splenocytes and seven of the clones could upregulate the expression of CD86 on mice B cells.Therefore,the CD40 monoclonal antibodies we generated can be used to test the expression of CD40,and to block or activate the CD40 signal.