A shear stress micromodel of urinary tract infection by the Escherichia coli producing Dr adhesin

Author summary Uropathogenic E. coli (UPEC) equipped with Dr fimbriae are associated with recurrent and chronic urinary tract infections (UTIs). The fimbriae assembled by the chaperone-usher pathway provide strong host-specific adherence which is, however, strongly modulated by the dynamically changing urine flow in the urinary tract (UT). In this paper, we use a dynamic in vitro micro-model of UTI to analyze the UT segment-specific impact of urinary outflow on the persistence and spread of Dr+ E. coli during host colonization. We conclude that the adhesive envelope formed by Dr fimbriae promotes strong and irreversible multivalent adherence of Dr+ E. coli to host receptors under flow conditions. We also observed that budding host cells-a model of any form of epithelial roughness, including carcinogenesis or physical injuries-facilitate the adherence of bacteria at flow conditions typically found in the UT, and our numerical simulations provided a mechanistic explanation for this effect. Finally, we combined the results to propose a rolling-shedding-refilling colonization model that shows how the wash off of detached colonized host cells may provoke a massive spread of UPEC. Our findings shed new light on UTI development and may be instrumental in the development of novel therapeutics. In this study, we established a dynamic micromodel of urinary tract infection to analyze the impact of UT-segment-specific urinary outflow on the persistence of E. coli colonization. We found that the adherence of Dr+ E. coli to bladder T24 transitional cells and type IV collagen is maximal at lowest shear stress and is reduced by any increase in flow velocity. The analyzed adherence was effective in the whole spectrum of physiological shear stress and was almost irreversible over the entire range of generated shear force. Once Dr+ E. coli bound to host cells or collagen, they did not detach even in the presence of elevated shear stress or of chloramphenicol, a competitive inhibitor of binding. Investigating the role of epithelial surface architecture, we showed that the presence of budding cells-a model microarchitectural obstacle-promotes colonization of the urinary tract by E. coli. We report a previously undescribed phenomenon of epithelial cell rolling-shedding colonization, in which the detached epithelial cells reattach to the underlying cell line through a layer of adherent Dr+ E. coli. This rolling-shedding colonization progressed continuously due to refilling induced by the flow-perturbing obstacle. The shear stress of fluid containing free-floating bacteria fueled the rolling, while providing an uninterrupted supply of new bacteria to be trapped by the rolling cell. The progressive rolling allows for transfer of briefly attached bacteria onto the underlying monolayer in a repeating cascading event.