利用小鼠感染模型筛选与鉴定幽门螺杆菌外膜蛋白抗原

Antigen screening and identification of Helicobacter pylori outer membrane proteins from murine infection model

目的:利用小鼠感染模型筛选与鉴定幽门螺杆菌SS1株的外膜蛋白抗原。方法:提取SS1株的外膜蛋白进行双向电泳,用幽门螺杆菌感染的小鼠血清作免疫印迹实验,将阳性反应蛋白点进行质谱鉴定分析,将肽质量指纹谱数据输入互联网上的蛋白质数据库进行检索。结果:获得32种抗原相关蛋白。通过与已有报道的幽门螺杆菌感染人抗原比较分析,发现大部分典型的保护性抗原在本实验中都可以检测到。结论:幽门螺杆菌感染的小鼠模型适用于人用保护性幽门螺杆菌抗原的筛选;而且此研究中得到的相关抗原蛋白对于寻找与鉴定幽门螺杆菌未知保护性抗原也有参考价值。

Objective：To screen antigen of Helicobacter pylori outer membrane proteins by murine infection model. Methods：Parallel two-dimensional gel electrophoresis （2D） of outer membrane proteins extracted from Helicobacter pylori strain SS1 was performed. Western blot of a duplicate 2D gel hybridized with serum from H. pylori-infected murine was employed. Immunogenic H. priori proteins identified in this way were digested in gel by trypsin and the mass of generated peptides were measured by matrix assisted laser desorption ionization time of flight mass spectrometry （MALDI-TOF-MS）. The data obtained from peptide mass finger-printing （PMF） were searched using the internet available database. Results： 32 proteins were identified and they are in good agreement with typical protective antigens which reacted with serum from H. pylori-infeeted patients. Conclusion：The results suggest that murine model of H. priori may be valid to screen antigens for hmnan vaccination and the proteins identified in this paper are valuable for the selection of H. priori protective antigens as well.