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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 ,这些短肽分子能诱导一定程度的保护性免疫
Objective To screen out the mimic target epitopes of the natural resistance to schistosoma japonicum in mice by phage random 12-peptide library and to explore its immunoprotective effect against Schistosoma japonicum. Methods The phage 12 peptide library was screened by using the crude serum IgG as the solid phase ligand and screened according to the elution elution amplification. The specificity of the phage clones was tested by ELISA and the specificity Two clones were sequenced. Mice were immunized with mixed phage clones three times at 0, 1, and 3 weeks respectively. In the fifth week, each mouse was infected with 40 ± 1 Schistosoma japonicum cercaria via abdomen, , Counting the number of worms and the number of liver eggs. Results After three rounds of screening, the specific binding phage enrichment increased by 200 times. Twenty randomly selected phage clones were identified by ELISA. Nineteen clones were able to react specifically with rat serum. The sequences of the two clones sequenced by automatic sequencer were not homologous to the known sequences in GenBank. Compared with the control group, the mice immunized with mixed phage clones had an anti-worm rate of 33.6% and an egg reduction rate of 5.8%. Conclusions The use of phage display technology to obtain natural anti-schistosome resistance short peptides, these short peptide molecules can induce a degree of protective immunity