抗A型产气荚膜梭菌α毒素单克隆抗体杂交瘤细胞系的建立与中和效应  

Establishment of Hybridoma Cell Lines Secreting Monoclonal Antibodies against Alpha toxin of Clostridium perfringens Type A and Netralization Effect of Produced McAbs

作　　者：许崇波[1] 马从林 赵宝华[2] 朱平[1] 

机构地区：[1]解放军农牧大学军事兽医研究所,吉林长春130062 [2]河北师范大学生物系,河北石家庄050016

出　　处：《中国兽医学报》1999年第5期464-466,共3页Chinese Journal of Veterinary Science

基　　金：全军医药卫生科研基金

摘　　要：用提取的 Ａ 型产气荚膜梭菌 α毒素包涵体免疫 Ｂ Ａ Ｌ Ｂ／ｃ 小鼠后， 取小鼠脾细胞与 Ｓ Ｐ２／０ 骨髓瘤细胞进行融合和克隆化，经间接 Ｅ Ｌ Ｉ Ｓ Ａ 筛选，共获得 １ Ａ８、１ Ｃ３、１ Ｄ５、１ Ｄ８、１ Ｆ１、１ Ｈ１ 和 ２ Ｅ３ ７ 株稳定分泌单克隆抗体（ Ｍ ｃ Ａｂ）的杂交瘤细胞株。经鉴定，７ 株 Ｍ ｃ Ａｂ 的 Ｉｇ 亚类有 Ｉｇ Ｇ１（１ Ｄ８）、 Ｉｇ Ｇ３（１ Ａ８、１ Ｃ３ 和 ２ Ｅ３）和 Ｉｇ Ｍ（１ Ｄ５、１ Ｆ１ 和 １ Ｈ１）。细胞培养上清和腹水抗体效价分别为 １∶５１２～１∶１ ０２４ 和 １∶１０６ ～１∶１０８ 。尤为重要的是，２ Ｅ３ 杂交瘤细胞株分泌的 Ｍ ｃ Ａｂ 不仅能够中和 α毒素的磷脂酶 Ｃ活性和溶血活性，而且能够对致死性腹腔感染小鼠产生良好的被动保护作用。The spleen cells taken from BALB/c mice immunized with purified inclusion bodies of alpha toxin of Clostridium perfringens were fused with Sp2/0 myeloma cells. Seven hybridoma cell lines steadily secreted monoclonal antibodies were obtained by ELISA. These hybridoma cell lines were designated 1A8, 1C3, 1D5, 1D8, 1F1, 1H1 and 2E3 respectively, and their isotype subclass were IgG1(1D8), IgG3(1A8, 1C3 and 2E3) and IgM(1D5, 1F1 and 1H1). The ELISA titers of cell supernatant and ascites were 1∶512 1∶1024 and 1∶106 1∶108 respectively. Especially, monoclonal antibodies secreted by 2E3 hybridoma cell line not only could neutralize phospholipase C and hemolytic activities of alpha toxin, but could provide protection for mice from the challenge of lethal dose of alpha toxin of Clostridium perfringens type A.

关 键 词：A型产气荚膜梭菌 Α毒素 单克隆抗体 中和效应 杂交细胞株 

分 类 号：S852.616.3[农业科学—基础兽医学]