Uropathogenic Escherichia coli outer membrane antigens expressed during urinary tract infection
Uropathogenic Escherichia coli outer membrane antigens expressed during urinary tract infection
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DOI：
10.1128/iai.00337-07
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发表时间：
2007-08-01
影响因子：
3.1
通讯作者：
Mobley, Harry L. T.
中科院分区：
文献类型：
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作者：
Hagan, Erin C.;Mobley, Harry L. T.
由尿路致病性大肠埃希菌(UPEC)引起的单纯性尿路感染(UTI)是一种流行且潜在严重的传染病。在这项研究中，我们描述了一种免疫蛋白质组学方法在疫苗开发中的应用，该方法已成功用于识别其他病原菌的疫苗靶标。在模拟尿路环境的条件下，包括铁限制、渗透应激、人尿和暴露于尿路上皮细胞的条件下，分离肾盂肾炎E.ColiCFT073菌株的外膜。为了确定在实验性UTI期间引起体液反应的抗原，用双向凝胶电泳法分离外膜蛋白，并用20只慢性感染E.ColiCFT073的CBA/J小鼠的混合抗血清进行探测。总共有23个外膜抗原，包括一种新的铁化合物受体，与抗血清发生反应，并经质谱仪鉴定。这些抗原还包括已知在UPEC发病机制中起作用的蛋白，如Chua、Iron、IREA、Iha、Iuta和Flc。这些数据表明，在尿路感染期间，抗体反应是针对这些毒力相关因子的。我们还表明，编码Chua、Fe、假设蛋白c2482和Iuta的基因在UPEC菌株中显著高于粪便共生大肠杆菌分离株(P<；0.01)。因此，我们认为这项研究中确定的保守外膜抗原可能是UTI疫苗的合理候选者，该疫苗旨在诱导对UPEC感染的保护性免疫。
Uncomplicated urinary tract infection (UTI) caused by uropathogenic Escherichia coli (UPEC) represents a prevalent and potentially severe infectious disease. In this study, we describe the application of an immunoproteomics approach to vaccine development that has been used successfully to identify vaccine targets in other pathogenic bacteria. Outer membranes were isolated from pyelonephritis strain E. coli CFT073 cultured under conditions that mimic the urinary tract environment, including iron limitation, osmotic stress, human urine, and exposure to uroepithelial cells. To identify antigens that elicit a humoral response during experimental UTI, outer membrane proteins were separated by two-dimensional gel electrophoresis and probed using pooled antisera from 20 CBA/J mice chronically infected with E. coli CFT073. In total, 23 outer membrane antigens, including a novel iron compound receptor, reacted with the antisera and were identified by mass spectrometry. These antigens also included proteins with known roles in UPEC pathogenesis, such as ChuA, IroN, IreA, Iha, IutA, and FliC. These data demonstrate that an antibody response is directed against these virulence-associated factors during UTI. We also show that the genes encoding ChuA, IroN, hypothetical protein c2482, and IutA are significantly more prevalent (P < 0.01) among UPEC strains than among fecal-commensal E. coli isolates. Thus, we suggest that the conserved outer membrane antigens identified in this study could be rational candidates for a UTI vaccine designed to elicit protective immunity against UPEC infection.