A high‐throughput assay for the measurement of uropathogenic Escherichia coli attachment to urinary bladder cells

Abstract

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.

Depletion of multidrug‐resistant uropathogenic Escherichia coli BC1 by ebselen and silver ion

“…UPEC adheres to bladder epithelial cells in a type I pili‐dependent manner and enters epithelial cells, 41,42 forming intracellular bacterial communities (IBCs) 43 . Their ability to attach and enter into cells in the bladder is a limiting step for their pathogenicity 44 45 the UPEC subsets exist as residual IBCs with resisting neutrophils and escape from IBS, and then invade the new bladder epithelial cells 46 .…”