中国寄生虫学与寄生虫病杂志 ›› 2000, Vol. 18 ›› Issue (4): 7-219.

• 论著 • 上一篇    下一篇

旋毛虫部分抗原表位的识别与分析

詹艳爱1;严自助2;王文2;吕再婴2;朱振勤1
  

  1. 华东师范大学生物系!上海200062;中国预防医学科学院寄生虫病研究所!上海200025
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2000-08-30 发布日期:2000-08-30

LOCALIZATION AND CHARACTERIZATION OF PARTIAL IMMUNODOMINANT ANTIGEN EPITOPES OF TRICHINELLA SPIRALIS

ZHAN Yan-ai 1;YAN Zi-zhu 2;WAN Wen 2;LU Zai-ying 2;ZHU Zhen-qin 1   

  1. 1 Department of Biology East China Normal University;shanghai 200062; 2 Institute of Parasitic Diseases;Chinese Academy of Preventive Medicine;Shanghai 200025
  • Received:1900-01-01 Revised:1900-01-01 Online:2000-08-30 Published:2000-08-30

摘要:   [目的 ]筛选旋毛虫肌幼虫可溶性抗原中具有免疫显性的表位。 [方法 ]采用杂交瘤技术 ,获得 15株特异性单克隆抗体 ,随后用酶联免疫吸附试验 (ELISA)、免疫印迹法 (Westernblotting)和间接免疫荧光试验(IFA)对部分免疫显性抗原进行分析。 [结果 ]Westernblotting试验显示 ,6株单抗与旋毛虫肌幼虫可溶性抗原反应显示有特异条带 ,分子量为 40~ 70kDa ;而多抗血清则可识别 2 0~ 2 0 0kDa之间 10条条带。IFA可观察到 ,6株单抗中有 4株单抗的靶抗原定位在旋毛虫肌幼虫表皮层上 ,另 2株定位于杆状体 (stichosome)及表皮层。 [结论 ]识别与分析部分旋毛虫肌幼虫可溶性抗原中具有免疫显性的表位 ,为纯化旋毛虫的抗原及疫苗靶抗原的研制提供了有价值的实验依据。

关键词: 旋毛虫, 表位, 单克隆抗体, 免疫印迹法, 免疫荧光

Abstract:  Objective] To screen and characterize immunodominant antigen epitopes on the soluble antigens of Trichinella spiralis (T s ) . [Methods]15 monoclonal antibodies (McAbs) against T s muscle larva(ML) soluble antigens were obtained by using hybridoma technique. The reactivity of monoclonal and polyclonal antibodies were tested by ELISA, Western blotting and indirect immunofluorescence assay(IFA). [Results] The Western blotting result showed that of the 15 McAbs, 6 could bind to the T s ML antigens displaying molecular weights of 40~70 kDa. Polyclonal sera could react with more than 10 bands having molecular weights of 20~200 kDa. Among the 6 McAbs, 4 could recognize epitopes on the cuticle surface and the other two could recognize epitopes on both the cuticle surface and the stichosome. [Conclusion]The antigen epitopes of T s recognized by 6 McAbs had been characterized.

Key words: Trichinella spiralis, epitope, monoclonal antibody, Western blotting, immunofluoscence