title: Assessment of virulence of uropathogenic Escherichia coli type 1 fimbrial mutants in which the invertible element is phase-locked on or off.
abstract: Type 1 fimbria is a proven virulence factor of uropathogenic Escherichia coli (UPEC), causing urinary tract infections. Expression of the fimbria is regulated at the transcriptional level by a promoter situated on an invertible element, which can exist in one of two different orientations. The orientation of the invertible element that allows the expression of type 1 fimbriae is defined as "on," and the opposite orientation, in which no transcription occurs, is defined as "off." During the course of a urinary tract infection, we have observed that the infecting E. coli population alternates between fimbriated and nonfimbriated states, with the fimbriated on orientation peaking at 24 h. We propose that the ability of the invertible element to switch orientations during infection is itself a virulence trait. To test this hypothesis, nucleotide sequence changes were introduced in the left inverted repeat of the invertible element of UPEC pyelonephritis strain CFT073 that locked the invertible elements permanently in either the on or the off orientation. The virulence of these mutants was assessed in the CBA mouse model of ascending urinary tract infection at 4, 24, 48, and 72 h postinoculation (hpi). We conducted independent challenges, in which bladders of mice were inoculated with either a single mutant or the wild type, and cochallenges, in which a mutant and the wild type were inoculated together to allow direct competition in the urinary tract. In both sets of experimental infections, the locked-off mutant was recovered from the urine, bladder, and kidneys in significantly lower numbers than the wild type at 24 hpi (P < or = 0.05), demonstrating its attenuation. Conversely, the locked-on mutant was recovered in higher numbers than the wild type at 24 hpi (P < or = 0.05), showing enhanced virulence of this mutant. No significant differences were seen between the mutants and wild type in the urine or the bladder at 48 or 72 hpi. However, the wild type outcompeted the locked-off mutant in the kidneys during the cochallenge experiment at 72 hpi (P = 0.009). Overall, these data suggest that the ability of the invertible element controlling type 1 fimbria expression to phase vary contributes significantly to virulence early (24 hpi) in the course of a urinary tract infection by UPEC and most profoundly influences colonization of the bladder.
authors.affiliation: Department of Microbiology and Immunology, University of Maryland School of Medicine, Baltimore, Maryland 21201, USA.
authors.forename: Nereus W
authors.initials: NW
authors.lastname: Gunther
authors.forename: Jennifer A
authors.initials: JA
authors.lastname: Snyder
authors.forename: Virginia
authors.initials: V
authors.lastname: Lockatell
authors.forename: Ian
authors.initials: I
authors.lastname: Blomfield
authors.forename: David E
authors.initials: DE
authors.lastname: Johnson
authors.forename: Harry L T
authors.initials: HL
authors.lastname: Mobley
mesh_terms: D000818:Animals; D004269:DNA, Bacterial / Q000502:physiology*; D004195:Disease Models, Animal; D004926:Escherichia coli / Q000235:genetics* / Q000254:growth & development / Q000472:pathogenicity*; D004927:Escherichia coli Infections / Q000382:microbiology; D005260:Female; D010861:Fimbriae, Bacterial / Q000235:genetics*; D005838:Genotype; D051379:Mice; D008808:Mice, Inbred CBA; D016296:Mutagenesis; D010641:Phenotype; D012045:Regulatory Sequences, Nucleic Acid / Q000502:physiology*; D014552:Urinary Tract Infections / Q000382:microbiology; D014774:Virulence
chemical_list: D004269:DNA, Bacterial