García Méndez Karellen Beren, Bragagnolo Gabriel, O'Callaghan David, Lavigne Jean-Philippe, Keriel Anne
U1047, UFR de Médecine, Inserm, Nîmes, Cedex, France.
U1047, Université de Montpellier, Nîmes, Cedex, France.
Int J Exp Pathol. 2016 Apr;97(2):194-201. doi: 10.1111/iep.12181. Epub 2016 Jun 8.
Strains of uropathogenic Escherichia coli (UPEC) are the major causative agent of urinary tract infections (UTI), the most common infectious diseases in the world. Their ability to attach and enter into cells in the urinary tract is a limiting step for their pathogenicity. Many studies are thus focussing on these key mechanisms to propose new therapeutic strategies. To facilitate such studies, we developed a fast and high-throughput assay which makes it possible to monitor the interaction of UPEC with cultured human uroepithelial cells. This assay allows measurement of the in vitro association of fluorescently labelled clinical isolates with bladder epithelial cells using flow cytometry in a microplate format. The assay was sensitive enough to detect variations between isolates expressing different adhesins and virulence factors and the inhibitory effect of proanthocyanidins. Thus we have developed a fast and robust assay which allows us to measure variations in the adhesion properties of UPEC to human bladder cells. This novel assay will be valuable for the study of initial steps of pathogenesis in UTI and for the screening or validation of inhibitory molecules.
尿路致病性大肠杆菌（UPEC）菌株是尿路感染（UTI）的主要病原体，而尿路感染是世界上最常见的传染病。它们在尿路中附着并进入细胞的能力是其致病性的一个限制步骤。因此，许多研究都集中在这些关键机制上，以提出新的治疗策略。为了促进此类研究，我们开发了一种快速且高通量的检测方法，该方法能够监测UPEC与培养的人尿道上皮细胞之间的相互作用。该检测方法允许使用微孔板形式的流式细胞术测量荧光标记的临床分离株与膀胱上皮细胞的体外结合。该检测方法灵敏度足够高，能够检测表达不同粘附素和毒力因子的分离株之间的差异以及原花青素的抑制作用。因此，我们开发了一种快速且可靠的检测方法，使我们能够测量UPEC对人膀胱细胞粘附特性的变化。这种新颖的检测方法对于研究UTI发病机制的初始步骤以及筛选或验证抑制分子将具有重要价值。
