用噬菌体展示技术筛选日本血吸虫免疫原的模拟短肽  

Screening of the mimic epitope of antigen from Schistosoma japonicum by using phage display techniques

作　　者：王敏[1] 易新元[1] 曾宪芳[1] 周东明[1] 张顺科[1] 章洁[1] 李忠杰[1] 王庆林[1] 袁仕善[1] 

机构地区：[1]中南大学湘雅医学院血吸虫病研究室,长沙410078

出　　处：《中华微生物学和免疫学杂志》2002年第3期295-298,共4页Chinese Journal of Microbiology and Immunology

摘　　要：目的　从噬菌体随机 12肽库免疫筛选出大鼠对血吸虫天然抗性的模拟靶表位 ,探讨其抗日本血吸虫的免疫保护效果。方法　用粗提大鼠血清IgG作固相配体筛选噬菌体 12肽库 ,按吸附 洗脱 扩增的淘洗过程进行 3轮筛选 ;随机挑取噬菌体克隆用ELISA检测其特异性 ,并对其中的 2个克隆进行测序 ;用混合噬菌体克隆免疫小鼠 3次 ,分别在 0、1、3周进行 ,第 5周每鼠经腹部感染 4 0± 1条日本血吸虫尾蚴 ,4 5d后剖杀冲虫 ,计虫数及肝卵数。结果　经 3轮筛选 ,特异性结合噬菌体富集增加了 2 0 0多倍 ,随机挑取 2 0个噬菌体克隆经ELISA鉴定 ,有 19个克隆能与大鼠血清呈特异性反应。自动测序仪测序的 2个克隆的序列与GenBank的已知序列无同源性。与对照组相比 ,混合噬菌体克隆免疫小鼠的减虫率为 33.6 % ,减卵率为 5 9.8%。结论　利用噬菌体展示技术可获得天然抗血吸虫抗性的短肽 。Objective To obtain short peptides simulating antigenic epitopes related with natural resistance to Schistosoma japonicum (S.j ) in rats, and to explore their immune protective immunity against S.j in mice. Methods Phage random peptide library of 12 amino acids was immunoscreened with purified IgG from sera of rats. Positive clones which were obtained after three rounds of biopanning were detected by ELISA, and two of them were sequenced. Kunming mice were immunized with mixed phage clones for three times. Each mouse challenged about 40 S.j cercaria, and all mice were perfused after 45 days. The worms and the liver eggs were counted. Results The specific phages binding to IgG were enriched 200 folds after three rounds of biopanning. 20 clones were detected by ELISA, 19 of them bound to the specific IgG from rats′ sera. The sequences of two clones showed no homology with other sequences in the GenBank. Compared with the control groups. The reduction rates of the worm burden and liver egg were 33.6% and 59.8%, respectively. Conclusion The specific peptides simulating antigenic molecules with natural resistance to S.j in rats can be obtained by immunosceening phage random peptide library and a protective immunity against S.j can be stimulated by these epitopes in mice. [

关 键 词：日本血吸虫 模拟短肽 免疫筛选 噬菌体随机肽库 

分 类 号：R392[医药卫生—免疫学]