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实验性尿路感染期间大肠杆菌1型菌毛表达及相变的体内检测

In vivo detection of Escherichia coli type 1 fimbrial expression and phase variation during experimental urinary tract infection.

作者信息

Struve C, Krogfelt K A

机构信息

Department of Gastrointestinal Infections, Statens Serum Institut, Copenhagen, Denmark.

出版信息

Microbiology (Reading). 1999 Oct;145 ( Pt 10):2683-90. doi: 10.1099/00221287-145-10-2683.

Abstract

Adhesion mediated by fimbriae is thought to play an important role in the pathogenesis of urinary tract infections (UTI) by Escherichia coli. The majority of clinical isolates of E. coli from UTI are able to express type 1 fimbriae. However, the importance of these fimbriae as a virulence factor has been controversial. To investigate the expression of type 1 fimbriae in vivo during UTI, mice were transurethrally infected with uropathogenic E. coli C175-94 and type 1 fimbrial expression was determined directly by two independent methods at 2 h, 1 d and 3 d after infection. By use of an assay combining in situ rRNA hybridization and immunofluorescence, all bacterial cells detected in urine, bladders and kidneys from mice sacrificed 1 and 3 d after onset of infection were found to express type 1 fimbriae. In contrast, the majority of cells in the suspension used for infection of mice and specimens from mice sacrificed 2 h after inoculation were found to be non-fimbriated. Similar results were obtained with a PCR assay revealing the orientation of the invertible promoter driving the transcription of type 1 fimbrial genes. Whilst the promoter in both ON and OFF positions could be amplified from the suspension used for infection and specimens from mice sacrificed 2 h after inoculation, at 1 and 3 d after onset of infection only the promoter in the ON orientation could be amplified. These results show that introduction of E. coli C175-94 into the mouse urinary tract leads to markedly enhanced expression of type 1 fimbriae.

摘要

由菌毛介导的黏附被认为在大肠杆菌引起的尿路感染(UTI)发病机制中起重要作用。大多数从UTI患者分离出的临床大肠杆菌菌株能够表达1型菌毛。然而,这些菌毛作为一种毒力因子的重要性一直存在争议。为了研究UTI期间体内1型菌毛的表达情况,将小鼠经尿道接种致病性大肠杆菌C175 - 94,在感染后2小时、1天和3天通过两种独立方法直接测定1型菌毛的表达。通过使用原位rRNA杂交和免疫荧光相结合的检测方法,发现在感染开始后1天和3天处死的小鼠的尿液、膀胱和肾脏中检测到的所有细菌细胞都表达1型菌毛。相比之下,用于感染小鼠的悬液中的大多数细胞以及接种后2小时处死的小鼠的标本中的细胞被发现没有菌毛。用PCR检测揭示驱动1型菌毛基因转录的可逆启动子的方向也得到了类似结果。虽然在用于感染的悬液和接种后2小时处死的小鼠的标本中都能扩增出处于ON和OFF位置的启动子,但在感染开始后1天和3天,只能扩增出处于ON方向的启动子。这些结果表明,将大肠杆菌C175 - 94引入小鼠尿道会导致1型菌毛表达明显增强。

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