幽门螺杆菌的免疫外膜蛋白质组学及其与胃部疾病的关系

目的：应用蛋白质组学技术研究幽门螺杆菌（HP）的候选抗原，用于胃部疾病临床诊断、治疗和疫苗开发。方法：选择自胃腺癌患者胃粘膜分离出且经确定的HP菌株（HP161），采用二维凝胶电泳（2-DE）分析外膜蛋白，并通过PDQuest图像分析软件分析蛋白质图谱，进行蛋白斑点检测和分析；收集20例HP感染患者和10例非HP感染相关性胃炎患者血清，采用免疫印迹技术分析HP161的外膜蛋白质与上述血清的反应性。结果：HP161有129个蛋白斑点；被慢性活动性胃炎和胃癌患者血清不同识别的特定抗原有10个；应用胃癌患者血清时，具有免疫反应性，而用胃炎患者血清时则无，等电点范围较宽，分子量多数在31kDa以下。结论：2-DE是研究HP外膜蛋白质组学的重要途径之一，并可能对其进行鉴定作为临床诊断的候选分子；经免疫蛋白组学检测的资料与公共数据库进行对比分析，以有助于寻找更具免疫原性的蛋白质标记物用于诊断分析和疫苗设计。

Immunoproteomics of Helicobacter pylori Outer Membrane Proteins and Relation to Gastric Diseases

Objective:To investigate whether H.pylori expresses disease-specific proteins that can be detected by two-dimensional polyacrylamide gel electrophoresis (2-D PAGE).A systematic, proteome based approach was chosen to detect candidate antigens of H.pylori for diagnosis, therapy and vaccine development.Methods:H.pylori strain HP161 isolated from gastric cancer was separated and analyzed by 2-D PAGE.Sera from patients with active H.pylori infection(including 10 cases with chronic active gastritis and 10 with gastric cancer)and a control group with unrelated gastric disorders (n=10)were collected and analyzed by Western blotting for the reactivity against the outer membrane proteins (OMP)of HP161.Results:2-DE gels resolved 129 proteins spots in HP161.10 antigens were recognized differently by sera from chronic gastritis and gastric cancer patients, which may identify them as candidate indicators for clinical manifestations.The data from these immunoproteomic analyses were compared with the public database (http://www.mpiib-berlin.mpg.de/2D-PAGE).We propose that collection of new antigens such as those recognized here will be useful in serologic tests for detecting and monitoring ,H.pylori infection and may also serve as potential targets for antimicrobial agent or vaccine development.Conclusion:2-D PAGE is useful to separate and identify potential antigens of H.pylori.The comparison of the antigenic patterns recognized by these sera may reveal association of specific H.pylori antigens with antibodies in patients with particular gastric pathologies.