Flagella allow uropathogenic Escherichia coli ascension into murine kidneys - PubMed (original) (raw)

Flagella allow uropathogenic Escherichia coli ascension into murine kidneys

William R Schwan. Int J Med Microbiol. 2008 Jul.

Abstract

Uropathogenic Escherichia coli (UPEC) cause bladder and kidney infections in humans and mice. UPEC initiate many kidney infections by ascending out of infected bladders, but how this occurs is not well understood. To determine if the flagella were responsible for the ascension of UPEC to the kidneys, a fliC mutation in strain NU149 was created. The fliC mutant spread poorly on soft agar plates, and 12h post-inoculation of murine urinary tracts, ascension into the murine kidneys was compromised in this mutant strain compared with wild-type bacteria. Complementation of the mutation restored the ability to spread on soft agar plates and ascend into the murine kidneys. To confirm the fliC mutant results, an anti-flagella monoclonal antibody that has been previously described inhibited the spread of UPEC strain NU149 on soft agar plates. When the anti-flagella antibody was mixed with strain NU149 cells and the antibody-treated bacterial cells were used to infect mice, significantly fewer mice had kidney infections than mice that were injected with strain NU149 cells mixed with normal mouse serum or anti-type 1 pili antibody. These results suggest that E. coli flagella may be of importance in allowing the bacteria to ascend from the bladder and initiate kidney infections in humans, and the use of an antibody against the flagella could prevent the spread of UPEC into the kidneys.

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Figures

Fig. 1

Fig. 1

Soft agar motility assays of E. coli strain NU149. (A) Comparison of E. coli strain NU149 (black) to a fliC mutant strain (NU149 fliC 2; striped), or a complemented fliC mutant strain (NU149 fliC 2/pWS15; white). (B) Comparison of E. coli strain NU149 and the addition of PBS (black), monoclonal antibody 2DE1B7 (anti-flagella; striped), or monoclonal antibody C3C (anti-type 1 pili; white). Spread of the bacteria was measured in mm after 6 h incubation at 37°C. The results are the mean ± standard deviation of three separate runs.

Fig. 1

Fig. 1

Soft agar motility assays of E. coli strain NU149. (A) Comparison of E. coli strain NU149 (black) to a fliC mutant strain (NU149 fliC 2; striped), or a complemented fliC mutant strain (NU149 fliC 2/pWS15; white). (B) Comparison of E. coli strain NU149 and the addition of PBS (black), monoclonal antibody 2DE1B7 (anti-flagella; striped), or monoclonal antibody C3C (anti-type 1 pili; white). Spread of the bacteria was measured in mm after 6 h incubation at 37°C. The results are the mean ± standard deviation of three separate runs.

Fig. 2

Fig. 2

Soft agar motility assay using E. coli strain NU149 and the addition of (A) PBS, (B) monoclonal antibody 2DE1B7 (anti-flagella), and (C) monoclonal antibody C3C (anti-type 1 pili).

Fig. 3

Fig. 3

Independent challenges of mice with E. coli strain NU149, a fliC mutant, and a complemented fliC mutant strain. The E. coli strain NU149 (group 1, closed diamond), NU149 fliC 2 mutant strain (group 2, closed square), or strain NU149 fliC 2/pWS15 complemented with the pWS15 plasmid (group 3, open triangle) were individually inoculated into the bladders of female BALB/c mice. After 12 h, the kidneys were harvested to determine bacterial concentrations. Each data point represents the log10 CFU/ml per one mouse. Horizontal bars represent the median values of the populations.

Fig. 4

Fig. 4

Independent challenges of mice with E. coli strain NU149 mixed with antibody. The E. coli bacteria were mixed with normal mouse serum (group 1, closed diamond), 2DE1B7 anti-flagella antibody (group 2, closed square), or anti-type 1 pili antibody (group 3, open triangle) and then individually inoculated transurethrally into the bladders of female BALB/c mice. After 24 h, the kidneys were harvested to determine bacterial concentrations. Each data point represents the log10 CFU/ml per one mouse. Horizontal bars represent the median values of the populations.

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