A high-throughput assay for the measurement of uropathogenic Escherichia coli attachment to urinary bladder cells.
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.
U1047, UFR de Médecine, Inserm, Nîmes, Cedex, France.|U1047, Université de Montpellier, Nîmes, Cedex, France.
Karellen Beren
KB
García Méndez
U1047, UFR de Médecine, Inserm, Nîmes, Cedex, France.|U1047, Université de Montpellier, Nîmes, Cedex, France.
Gabriel
G
Bragagnolo
U1047, UFR de Médecine, Inserm, Nîmes, Cedex, France.|U1047, Université de Montpellier, Nîmes, Cedex, France.
David
D
O'Callaghan
U1047, UFR de Médecine, Inserm, Nîmes, Cedex, France.|U1047, Université de Montpellier, Nîmes, Cedex, France.|Service de Microbiologie, CHU Carémeau, Nîmes, Cedex, France.
Jean-Philippe
JP
Lavigne
U1047, UFR de Médecine, Inserm, Nîmes, Cedex, France.|U1047, Université de Montpellier, Nîmes, Cedex, France.
Anne
A
Keriel
D001422:Bacterial Adhesion / Q000187:drug effects / Q000502:physiology*; D001431:Bacteriological Techniques / Q000379:methods; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000382:microbiology / Q000502:physiology; D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D008517:Phytotherapy; D010936:Plant Extracts / Q000494:pharmacology; D001743:Urinary Bladder / Q000166:cytology / Q000382:microbiology*; D014552:Urinary Tract Infections / Q000382:microbiology*; D056689:Uropathogenic Escherichia coli / Q000472:pathogenicity / Q000502:physiology*; D029799:Vaccinium macrocarpon; D014774:Virulence
D010936:Plant Extracts
Escherichia coli; ExPEC; UPEC; adhesion; flow cytometry; screening; urinary tract infections