抗多房棘球绦虫原头节单克隆抗体的制备  

Production of A Monoclonal Antibody Specific to Protoscolex of Echinococcus multilocularis

作　　者：王昕[1,2] 刘巧凤[1] 路睿[1] 徐佳楠[1] 李调英[3] 孙磊[1] 张亚楼[2] 孙力杨[2] 张辉[2] 陈建平[1] 

机构地区：[1]四川大学华西基础与法医学院,成都610041 [2]新疆医科大学基础医学院,乌鲁木齐830054 [3]四川省疾病预防控制中心,成都610041

出　　处：《中国寄生虫学与寄生虫病杂志》2009年第4期328-331,共4页Chinese Journal of Parasitology and Parasitic Diseases

基　　金：国家自然科学基金(No.30660045)~~

摘　　要：目的制备抗泡球蚴组织单克隆抗体,并鉴定其特异性。方法运用泡球蚴组织粗抗原免疫小鼠制备免疫脾细胞,运用淋巴细胞杂交瘤技术将其与小鼠骨髓细胞SP2/0融合。采用ELISA和免疫组化方法筛选抗泡球蚴原头节单克隆抗体的杂交瘤细胞株。检测所获细胞株的染色体数目,采用ELISA和免疫组化法鉴定所获抗体的特异性。结果获得了一株能稳定分泌的抗泡球蚴原头节单克隆抗体的杂交瘤细胞株6G2A7F7。染色体数目为98条,为明显的融合细胞核型。所分泌的抗体McAbP325能与泡球蚴中的生发层和原头节特异结合,特别对原头节上的小钩和吸盘表现出很强的结合反应。但与棘球蚴(人源)和水泡带绦虫幼虫(细颈囊尾蚴)组织无特异性结合反应。结论制备的杂交瘤细胞能稳定分泌McAbP325,为泡球蚴的细胞分类、免疫组化和抗原研究提供工具和基础。Objective To prepare monoclonal antibody （McAb） specific to protoscolex of Echinococcus multil- ocularis. Methods BALB/c mice were immunized with crude antigen derived from E. multilocularis metacestodes. Spleen cells from immunized BALB/c mice were fused with SP2/0 myeloma cells by using hybridoma technique. ELISA and immunohistochemical staining were used to select hybridomas that secreted McAb P325 which especially against protoscolex. The number of metaphase chromosomes of hybridoma cells was counted. Characteristics of McAb P325 were identified by ELISA and immunohistochemical staining. Results One hybridoma cell clone secreting McAb against protoscolex was obtained. The number of metaphase chromosomes found in hybridoma cells was 98, which showed the characteristics of their parents. Immunohistochemical analysis showed that McAb P325 demonstrated binding activity to the germinal layer and protoscolex of E. multilocularis, especially to the hooklets and suckers, while did not bind with E. granulosus metacestodes and Cysticercus tenuicollis. Conclusion The McAb is a valuable tool for immunohistochemical analysis, cell classification of E. multilocularis protoscolex, and study of specific antigen.

关 键 词：多房棘球绦虫 泡球蚴包囊 原头节 单克隆抗体 免疫组化 

分 类 号：R383.33[医药卫生—医学寄生虫学]