摘要
目的 从噬菌体随机肽库中筛选出模拟旋毛虫特异性抗原表位的短肽分子 ,探讨其抗血吸虫的交叉免疫保护效果。 方法 以纯化的旋毛虫感染鼠血清IgG为配基 ,亲合筛选法富集特异性噬菌体 ,随机挑取噬菌体克隆用ELISA检测其特异性 ;混合噬菌体克隆经皮下免疫小鼠 3次 ,攻击感染后第 4 5天剖杀小鼠 ,观察减虫和减卵效果。 结果 经 3轮筛选 ,特异性噬菌体得到了有效的富集 ,第三轮洗脱噬菌体的产量约为第一轮的 15 0倍。随机挑取 2 4个噬菌体克隆经ELISA测定 ,有 2 1个克隆能与旋毛虫感染鼠血清IgG特异性反应。与对照组相比 ,混合噬菌体克隆免疫小鼠的减虫率与减卵率分别为 4 2 8%与 6 6 3% (P <0 0 0 1)。 结论 利用噬菌体随机肽库技术可获得模拟旋毛虫特异性抗原表位的短肽分子 ,这些短肽分子能诱导明显的抗血吸虫的保护性免疫。
Objective To obtain the short peptides mimicking antigenic epitopes of Trichinella spiralis ( T\^s\^ ), and explore their cross protective immunity against Schistosoma japonicum ( S\^j. ) in mice. Methods IgG antibodies were purified from sera of mice infected with T\^s\^ . The purified IgG was used to immunoscreen a phage random peptide library of 7 amino\|acid residues displayed as a fusion to protein of filamentous phage. Positive clones were obtained by affinity selection, the reactivity of each clone binding to specific IgG was detected by ELISA. Kunming mice were immunized subcutaneously three times with mixed phage clones. The mice were sacrificed 45 days after challenge. The worms and the liver eggs were counted. Results After three rounds of panning, the relevant phages had been enriched approximately 150 times in production as compared to those from the first round. Of 24 phage clones randomly selected from the third round biopanning, 21 clones were shown to actually bind to the specific IgG. As compared with the control group, the worm and the liver egg reduction rates in vaccination group were 42\^8% and 66\^3% ( P <0\^001), respectively. Conclusion The above results demonstrate that antigenic epitopes of T\^s\^ can be prepared by immunoscreening phage random peptide library and a significant protective immunity against S\^j. can be induced by these epitopes in mice.
出处
《中国寄生虫学与寄生虫病杂志》
CAS
CSCD
北大核心
2001年第5期268-271,共4页
Chinese Journal of Parasitology and Parasitic Diseases
关键词
日本血吸虫
旋毛虫
模拟表位
噬菌体随机肽库
Schistosoma japonicum,Trichinella spiralis ,mimic peptide,phage display library
