摘要
目的 在建立一种有成虫寄生但无虫卵肉芽肿形成的日本血吸虫感染小鼠及家兔新模型 ,并观察其具有较高的抗攻击感染保护力的基础上 ,用新模型兔血清筛选噬菌体随机 12肽库 ,并初步鉴定阳性噬菌体克隆诱导小鼠抗日本血吸虫感染的免疫保护作用。方法 用大肠杆菌抗原吸收后的新模型兔血清 ,经抗体捕获法筛选噬菌体 12肽库 ,经 3轮亲和筛选、富集后获得阳性多克隆。用常规 EL ISA法检测阳性多克隆噬菌体的抗原性。用滴度为 1× 10 1 4pfu的阳性多克隆噬菌体按0 - 2 - 4周方案皮下多点注射免疫小鼠 ,末次免疫 4周后 ,用日本血吸虫尾蚴攻击感染 ,同时设立常规感染兔血清筛获的阳性克隆免疫组、原始肽库免疫组、攻击感染对照组。结果 1新模型兔血清筛获的多克隆噬菌体 (滴度为 1× 10 1 4pfu)与新模型兔血清 (稀释度为 1∶ 4 0 0 )反应为强阳性 ,与常规感染兔血清 (1∶ 4 0 0 )反应为弱阳性 ,与正常兔血清 (1∶ 4 0 0 )反应为阴性。 2用新模型兔血清筛获的多克隆噬菌体、常规感染兔血清筛获的多克隆噬菌体及原始肽库分别免疫小鼠后诱导抗攻击感染的减虫率及每克肝脏减卵率分别为 2 7.2 %和 38.8%、17.8%和 35 .0 %、4 .5 %和 6 .0 %。结论 与用日本血吸虫常规感染模型兔血清筛获的多克隆噬菌体比较 。
Objective To screen the 12 mers-phage random peptide library using the serum from the new model rabbit and to identify the immuno-protection of the positive phages. The new model infected with Schistosoma japonicum was proved that has a high protection against the challenge infection. Methods After being absorbed by E.coli antibody, the serum of the new model rabbit was used to screen the peptide library. Through three rounds of biopaning and enriching, lots of positive phages were obtained and their antigenic ability was tested. Every mouse was immunized by subcutaneously injecting 1×10 14 pfu positive phages from the new model rabbit serum respectively at 0-2-4 th week. After 4 weeks of the last immunization, the challenge infection was performed. At the same time, several control groups including the group immunized with the phages from the rabbit serum of the normal model infected with Schistosoma japonicum, the group immunized with the original 12 mers-phage random peptide library and the control group of challenge infection were arranged. Results ①The positive clones of phage(1×10 14) from the new model rabbit serum were strongly recognized by the rabbit serum of the new model, weakly recognized by the rabbit serum of the normal model infected with Schistosoma japonicum,but not recognized by the serum of healthy rabbit. ②The reduction rate of adult worms and liver eggs induced by phages screened with the rabbit serum of the new model group and the nomal model group and that induced by the original peptide library were respectively 27 2% and 38 8 %, 17 8% and 35 0%, 4 5% and 6 0% Conclusion The new model group obtained a higher reduction rate of adult worms than the nomal model group (P<0 05), also obtained a obviously higher reduction rate of LEPG than original 12 mers-phage random peptide library (P<0 01)
出处
《中国血吸虫病防治杂志》
CAS
CSCD
2004年第2期90-94,共5页
Chinese Journal of Schistosomiasis Control
基金
国家自然科学基金项目 ( No. 3 0 2 40 0 74)
湖北省科技攻关项目 ( No.2 0 0 3 AA3 0 0 3 B0 4)
湖北省卫生厅科研基金重点项目 ( No.JX1A11)
关键词
日本血吸虫
噬菌体肽库
抗原表位
感染
免疫
Schistosoma japonicum
Peptide library
Epitope
Infection
Immunity
