Urogenit Tract Infect.  2017 Dec;12(3):122-129. 10.14777/uti.2017.12.3.122.

Adherence Assay of Uropathogenic Escherichia coli In Vivo and In Vitro

• Kim DY
• Lee JC

• KMID: 2400221
• DOI: http://doi.org/10.14777/uti.2017.12.3.122

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.

Keyword

Urinary tract infections; Virulence factors; Escherichia coli

MeSH Terms

Animals
Bacteria
Cell Culture Techniques
Cell Line
Colon
Cystitis
Escherichia coli
Humans
In Vitro Techniques*
Kidney
Mice
Microscopy, Electron, Scanning
Models, Animal
Pyelonephritis
Urinary Bladder
Urinary Tract Infections
Uropathogenic Escherichia coli*
Virulence
Virulence Factors
Virulence Factors

Figure

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  Fig. 1. Light micrographs showing 3 types of adherence pattern in HeLa, HEp-2, A-498, and J-82 cells. DA denotes a diffuse adherence to the TME104 strain on the dispersed surface of the cultured cells. LA represents a localized adherence of the TME303 strain to the localized areas of cultured cells and formed to particulate microcolonies. AA denotes an aggregative adherence of TME114 strain to the cultured cells. Note the characteristic aggregates of bacteria that showed “stacked-brick” appearance. They bound aggregately to both of cultured cells and bottom of cell culture dish (×500).

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  Fig. 2. Inflammation of the mouse bladder (A) and kidney (B) caused by bacterial infections. H&E, ×100.

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  Fig. 3. Scanning electron micrographs showing diffuse adherence (DA) of TME104, localized adherence (LA) of TME303, and aggregative adherence (AA) of TME114 Escherichia coli strains adhered to J-82 cells. Magnification, ×1,500.

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  Fig. 4. Scanning electron micrographs showing TME116 (A, B), TME303 (C, D), and TME114 (E, F) Escherichia coli strains adhering to mouse urinary bladder. Magnification, A, C, E: ×2,200; B, D, F: ×15,000. DA: diffuse adherence, LA: localized adherence, AA: aggregative adherence, NA: non-adherence.

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