抗人CD154单克隆抗体的构建及免疫学特性分析  

Preparation of Anti-hCD154 Monoclonal Antibody and Analysis of its Immunological Properties

作　　者：张春艳[1] 张志方[2] 宁波[1] 李树浓[3] 朱兆玲[1] 

机构地区：[1]中山医科大学免疫学教研室,广州510089 [2]中山医科大学生理学教研室,广州510089 [3]中山医科大学病理生理学教研室,广州510089

出　　处：《中国动脉硬化杂志》2001年第5期405-406,共2页Chinese Journal of Arteriosclerosis

摘　　要：为进一步研究人CD15 4单克隆抗体在抑制动脉粥样硬化发生和发展中的作用 ,采用重组人CD15 4分子免疫BALB c小鼠 ,用杂交瘤技术制备抗人CD15 4单克隆抗体 ,并分析其免疫学特性。结果发现 ,(1)有 6c5、1b6、5d3共三株杂交瘤细胞株的培养上清与GST hCD15 4融合蛋白反应 ,与GST不反应。 6c5、1b6和 5d3培养上清效价分别为 1∶12 8、1∶2 5 6和 1∶6 4。 (2 )经Western bolt鉴定 ,在分子质量为 5 6kDa处有一抗原抗体结合的条带 ,而分子质量为2 6kDa处无特异性条带。证明三株杂交瘤细胞株所分泌的单克隆抗体可以结合GST hCD15 4但不与GST反应 ,是特异性结合人CD15 4的单克隆抗体。 (3)流式细胞术发现 ,6c5细胞株分泌的单克隆抗体能与激活的人外周血T细胞结合。说明已构建的单克隆抗体是特异性针对人CD15 4的 ,为下一步用人CD15 4单克隆抗体抑制动脉粥样硬化的研究打下了基础。Aim To Prepare anti-hCD154 monoclonal antibody and analysis its immunological properties.Methods The GST-hCD154 fusion protein was used as antigen to immunize BALB/c mouse. B cell was fused with myeloma cell SP2/0 to produce hybridoma cell line that can secrete anti-hCD154 antibody. The hybridoma cells were identified by using ELISA, Western-blot and FACS method. Results (1) ELISA result demonstrated that culture supernatants of three hybridoma cells(6c5, 1b6, and5c3) can react with GST-hCD154 ,but not react with GST.(2) Western-blot show that three monoclonal antibodies could bound to a protein band with molecular weight of 56 kDa (GST-hCD154) ,but could not bound to a protein band with molecular weight of 26 kDa (GST). (3) FACS show that McAb 6c5 can bound to peripheral blood T cell activated. Conclusion 6c5 cell is a specific McAb against human CD154 molecules expressed on actived human T cells. Preparation of anti-hCD154 monoclonal antibody be used to study the inhibitory function of atherosclerosis progression.

关 键 词：人CD154 单克隆抗体 T淋巴细胞 动脉粥样硬化 

分 类 号：R543.503[医药卫生—心血管疾病] R392.11[医药卫生—内科学]