日本血吸虫辐照尾蚴模拟表位的筛选和初步鉴定

目的从噬菌体表面显示随机肽库中筛选日本血吸虫辐照尾蚴抗原模拟表位。方法用Sepharose4B亲和层析法,从日本血吸虫辐照尾蚴免疫鼠血清中纯化抗体,经正常血吸虫尾蚴抗原吸收后,用于12肽噬菌体随机肽库的免疫筛选。经三轮筛选,随机挑取20个噬菌体克隆,经DNA序列分析后,对各独立表位进行免疫学鉴定。结果DNA序列分析,获得4个有序列重复的噬菌体克隆;Dot-ELISA分析显示,4个克隆均能被辐照尾蚴免疫血清中IgM型抗体识别,反应强度有一定差别;P1和P2克隆还能被IgG型抗体识别。结论从噬菌体随机肽库中成功筛选到日本血吸虫辐照尾蚴抗原模拟表位。

Secreening and immunological identification of the mimic epitopes of the irradiated cercariae of Schistosoma japonicum

Objective To obtain the specific mimic epitopes of the irradiated cercariae of Schistosoma japonicum from the phage display library. Methods Sera from mice immunized with irradiated cercariae of Schistosoma japonicum were purified by Sepharose 4B conjugated with the irradiated cercariae antigens and absorbed with normal cercariae antigens of S. japonicum, then, it was used for immunoscreening of a phage display library of random peptide 12- mers. After 3 cycles of screening, DNA samples from 20 phage colnes were sequenced. The phage clones with different inserted epitopes were identified immunologically. Results 4 independent phage clones were determined. Dot- ELISA analysis showed that all of them could be recognized by specific IgM antibodies and P1 as well as P2 clones could also be recognized by specific IgG antibodies from the irradiated cercariae immunized sera. Conclusion The specific mimic epitopes of the irradiated cercariae of Schistosoma japonicum were screened successfully from the phage display library.