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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 screen short peptide molecules that mimic Trichinella specific epitopes from the phage random peptide library and investigate the cross-immunoprotective effect of anti-schistosomes. Methods Purified Trichinella spiralis-infected mouse serum IgG was used as ligand. The specific phage were enriched by affinity screening. The phage clones were randomly picked up and tested for specificity by ELISA. The mice were immunized subcutaneously with mixed phage clones for 3 times, Mice were sacrificed on day 45 to observe the effects of de-worming and egg reduction. Results After 3 rounds of screening, the specific phage was effectively enriched. The yield of the third round of phage was about 150 times of the first round. Twenty-four phage clones were randomly selected and tested by ELISA, and 21 clones were able to react specifically with serum IgG from Trichinella-infected mice. Compared with the control group, the mice immunized with mixed phage clones had the worm reduction rate and the egg reduction rate of 42.8% and 66.3%, respectively (P <0.01). Conclusion The phage random peptide library technology can be used to obtain short peptide molecules that mimic Trichinella specific epitopes, and these short peptide molecules can induce significant protective immunity against schistosome.