抗隐孢子虫单克隆抗体研制及其特异性的鉴定

中国寄生虫学与寄生虫病杂志 ›› 1998, Vol. 16 ›› Issue (4): 279-282.

抗隐孢子虫单克隆抗体研制及其特异性的鉴定

宗海红; 李为明; 张雪; 俞树荣

1 第三军医大学新桥医院儿科 2 第三军医大学微生物教研室

收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:1998-08-28 发布日期:1998-08-28

PREPARATION AND CHARACTERIZATION OF MONOCLONAL ANTIBODIES AGAINST CRYPTOSPORIDIUM PARVUM

Zong Haihong¹; Li Weiming ¹; Zhang Xue ²; Yu Shurong²

1 Department of Pediatrics; Xinqiao Hospital; Third Military Medical University; Chongqing 400037 2 Laboratory of Micrbiology, Third Military Medical University , Chongqing 400037

Received:1900-01-01 Revised:1900-01-01 Online:1998-08-28 Published:1998-08-28

摘要/Abstract

摘要： 目的：制备抗隐孢子虫单克隆抗体（McAb）并对其特异性进行鉴定。方法：用纯化的人源微小隐孢子虫卵囊免疫BALB／c小鼠，杂交瘤技术制备McAb，间接免疫荧光（IFA）和免疫印迹试验分析其特性。结果：制备出Z4C8、Z2B6、Z3D7和Z3D2四株能稳定分泌抗隐孢子虫McAb的杂交瘤细胞株。经鉴定，Z4C8和Z3D7为IgM ，Z3D2和Z2B6为IgG1，轻链均为κ链，与多种肠道病原体及肺孢子虫无交叉反应。除Z2B6在间接免疫荧光试验中识别卵囊壁外，其余均识别隐孢子虫子孢子（CSP）。免疫印迹定位表明，四株McAb分别识别42条CSP抗原中的20．5kDa、60．5kDa、33kDa和95kDa4个条带，其中Z4C8和Z3D7均识别20．5kDa主要抗原带。结论：这四株McAb在识别CSP抗原表位上具有不同的特性，但对隐孢子虫抗原均呈高度特异性反应。

关键词: 隐孢子虫, 单克隆抗体, 抗原表位

Abstract: AIM: To develop and characterize the monoclonal antibodies(McAbs) against
Cryptosporidium parvum (C.parvum). METHODS: BALB/c mice were immunized with purified oocyst
antigens of C.parvum from human isolates and McAbs against C.parvum were prepared by using
hybridoma technique. The characterization of McAbs was studied by means of indirect
immunofluorescent assay (IFA) and Western blot analysis. RESULTS: Four clones of hybridoma cell
lines secreting McAbs against oocyst antigens of C.parvum were obtained. The immunoglobulin subclasses of these four McAbs were identifid as IgM (Z4C8,Z3D7) and IgG1 (Z2B6, Z3D2) and their light chains were Kchain. IFA results showed that three McAb (Z4C8, Z3D7 and Z3D2) conjugates produced a diffuse fluorescence in the Cryptosporidium sporozoites (CSP) and one (Z2B6) produced local fluorescence in the oocystwall of C. parvum. Four McAbs could recognize 4 of 42 polypeptide antigen bands of CSP with molecular weights of 20.5 kDa, 33 kDa, 60.5 kDa and 95 KDa, among which both Z4C8 and Z3D7 could recognize 20.5 kDa band that was defined as a principal antigenic protein of CSP. CONCLUSION: These four McAbs possess their own characteristics of recognizing antigenic epitopes on CSP, but react specifically against antigens of C. parvum.

Key words: Crystosporidium, McAb, antigenic epitope

宗海红;李为明;张雪;俞树荣. 抗隐孢子虫单克隆抗体研制及其特异性的鉴定[J]. 中国寄生虫学与寄生虫病杂志, 1998, 16(4): 279-282.

ZongHaihong;LiWeiming;ZhangXue;YuShurong. PREPARATION AND CHARACTERIZATION OF MONOCLONAL ANTIBODIES AGAINST CRYPTOSPORIDIUM PARVUM[J]. Chinese Journal of Parasitology and Parasitic Diseases, 1998, 16(4): 279-282.

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