Adherence Assay of UropathogenicEscherichia coliIn Vivo and In Vitro

Abstract

Purpose: This study aims to investigate the specific pathogenic properties or virulent determinant characteristics of uropathogenic Escherichia coli (UPEC) as bacterial adherence in tissue culture cells in vitro and the pathogenicity in animal model in vivo. Materials and Methods: Thirty strains of E. coli were isolated from urine of patients with acute pyelonephritis. Four cell lines-HeLa cells, HEp-2 cells, A-498 cells, and J-82 cells-were used for bacterial adherence assay. Histologic examination and scanning electron microscopy examination of pyelonephritis or cystitis, which was caused by E. coli, in mice was performed. Results: Sixteen (53.3%) strains of E. coli appeared to adhere to at least one or more kinds of four cell lines, and seven strains were able to adhere to all four cell lines. All of the tested E. coli strains were adhered to the mouse bladder and kidneys. The number of bacteria colonized in the kidney was greater than that of bladders in the following 5 strains of E. coli: TME104, TME107, TME113, TME306, and TME119. There was no difference in the number of bacteria colonized in the bladder and kidneys in the aspects of adherence patterns to tissue culture cells. Conclusions: Although there was no best choice cell lines in the adherence assay to identify the adherence patterns, combined assays of in vitro cell culture and in vivo model of mouse urinary tract infection appeared to be efficient methods to investigate the role of bacterial adherence in the pathogenesis of UPEC.

Virulence behavior of uropathogenic Escherichia coli strains in the host model Caenorhabditis elegans

“…to HEp-2 cells was unlikely to be associated with mice uroepithelial adherence as reported by Kim and Lee (2017). Likewise in our study, UPEC strains adhere to HEp-2 cell monolayers in a similar extent but only ECP110 resulted able to colonize the worm gut.…”

“…In the epithelial cell line HEp‐2, the in vitro model utilized in this study, both ECP45 and ECP110 were able to adhere and invade to a extent and they were unable to survive in cell monolayers. Adherence capacity of E. coli to HEp‐2 cells was unlikely to be associated with mice uroepithelial adherence as reported by Kim and Lee (). Likewise in our study, UPEC strains adhere to HEp‐2 cell monolayers in a similar extent but only ECP110 resulted able to colonize the worm gut.…”

Effect of Mannan Oligosaccharides Extracts in Uropathogenic Escherichia coli Adhesion in Human Bladder Cells

“…Due to their exploitation in animal supplementation as prebiotics, scientific reports on MOS have focused on their impact on the gastrointestinal microflora and immune system of farm animals [ 36 , 37 , 38 , 39 , 40 ]. Studies on the effect of MOS in UTIs are scarce [ 41 ], with most reports focusing on the benefit of using D-mannose as a therapeutic agent against UTIs [ 25 , 41 , 42 , 43 , 44 , 45 ]. One of the mechanisms through which UPEC is capable of adhering to cells is mediated by a bacterial ligand specific to D-mannose (FimH) located at the tip of type 1 pili anchored to UPEC’s outer membrane [ 1 , 11 , 46 , 47 , 48 , 49 ].…”

“…Incubation periods were chosen considering that the majority of ingested mannose is filtered by the kidneys and excreted to the bladder via urine within 30 to 60 min [51] and that, although urinary frequency depends on a multitude of factors, it is commonly accepted that most Due to their exploitation in animal supplementation as prebiotics, scientific reports on MOS have focused on their impact on the gastrointestinal microflora and immune system of farm animals [36][37][38][39][40]. Studies on the effect of MOS in UTIs are scarce [41], with most reports focusing on the benefit of using D-mannose as a therapeutic agent against UTIs [25,[41][42][43][44][45]. One of the mechanisms through which UPEC is capable of adhering to cells is mediated by a bacterial ligand specific to D-mannose (FimH) located at the tip of type 1 pili anchored to UPEC's outer membrane [1,11,[46][47][48][49].…”

Genomic characterization of a multidrug-resistant uropathogenic <i>Escherichia coli</i> and evaluation of <i>Echeveria</i> plant extracts as antibacterials

“…Once the MIC of the extracts was determined, adherence assays were performed to determine whether the Echeveria subrigida extract could influence the initial step of UPEC infection. Adherence is essential for the establishment of a UTI and a triggering event for the subsequent steps of the pathogenic mechanism [77] . Currently, anti-adherence therapies offer an alternative to bacterial infections by blocking the interaction of bacteria with epithelial cells without killing them, thus eliminating the bacteria.…”