A model for ascending unobstructed urinary tract infection was developed in mice to study the pathogenesis of urinary tract infection induced by Escherichia coli associated with urinary tract infection in humans. Specifically, the model was designed to monitor the initial stages of the infectious process, e.g., bacterial adhesion. Mice were selected since the specificity and intensity of bacterial attachment of pyelonephritogenic E. coli strains to human and mouse uroepithelial cells were similar. Female mice were infected by urethral catheterization and installation of bacteria in the urinary bladder. To maximize clearance of unattached bacteria, no obstructive manipulations were performed. After sacrifice, the persistence of bacteria in kidneys and bladder was determined by viable counts on homogenized tissues. The experimental infection was standardized by using one pyelonephritis (HU734) and one normal fecal (414) E. coli isolate. With both strains all of the bladders became infected, but E. coli 414 was eliminated more rapidly than HU734. The percentage of positive kidney cultures increased with the bacterial inoculum concentration and volume. An inoculum of 0.05 ml containing 10(10) bacteria per ml was selected, giving the highest percentage of positive kidney cultures without detectable bacterial spread to the blood stream. The variation in the percentage of positive kidney cultures possibly depended on the degree of vesicoureteric reflux in the individual animals. Both in the kidneys and in the urinary bladders, strain HU734 yielded higher numbers of bacteria at 24 h and persisted longer than did strain 414. Several E. coli pyelonephritis isolates with properties associated with virulence in the human urinary tract consistently were recovered from mouse kidneys and bladders in higher numbers than E. coli strains of human fecal origin lacking those properties. The role of bacterial adhesion per se is the topic of the accompanying paper.

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Glycoconjugates containing the disaccharide unit GlcNAc beta 1 leads to 3Gal beta were suggested as receptors for pneumococci adhering to human pharyngeal epithelial cells. The receptor activity was detected both by inhibition of adhesion by an excess of free oligosaccharide and by induction or increase of adhesion after coating of target cells with glycolipid. Studies with free natural and synthetic oligosaccharides identified the disaccharide GlcNAc beta 1 leads to 3Gal beta as one critical binding site. The specificity of recognition was shown inter alia by the lack of inhibitory activity of GlcNAc beta 1 leads to 4Gal beta, which differs only in the linkage of the two sugars. Specific interference with pneumococcal adhesion by administration of soluble receptor sugar may improve our understanding of the role of adhesion in vivo.

The affinity of uropathogenic Escherichia coli to kidneys and bladders of experimentally infected mice was shown to be determined in part by the adhesive properties of the infecting bacteria. Mice were infected with various pairwise combinations of two homogeneic sets of bacteria: (i) mutants derived from a human pyelonephritis E. coli isolate which were selected to express either or both adhesins specific for globoseries glycolipid receptors or for “mannosides”; and (ii) transformants of a normal fecal isolate which harbored recombinant plasmids encoding the genes for one or the other adhesin or which harbored only the vector plasmid. The relative efficiency of survival of the strains to be compared was evaluated in each animal by plating on selective media of samples of homogenized kidneys and bladders taken 24 h after intravesical inoculation. The presence of adhesins specific for globoseries glycolipid receptors, which mediate the in vitro mannose-resistant attachment to human and mouse uroepithelial cells, enhanced bacterial recovery from both kidneys and bladders of infected animals. The addition to the infecting strain of adhesins binding mannoside residues further improved bacterial recovery from the bladder, but not from the kidney. The mutants and transformants with adhesins binding only mannosides were recovered in higher numbers from the bladder than those expressing adhesins specific for the globoseries glycolipids only. There was apparent selection in vivo decreasing expression of mannoside binding adhesins in the kidneys, but not in the bladders, of animals infected with the mutant expressing both types of adhesins. Regardless of adhesive properties, the mutants of the pyelonephritis isolate were recovered in significantly higher numbers than the fecal isolate with adhesins encoded on recombinant plasmids. We conclude that the adhesive properties in part determine the localization and retention of bacteria in the mouse urinary tract. However, the addition of adhesins to a commensal E. coli strain was not sufficient to confer colonization capacity comparable to that of a pyelonephritis strain.

The capacity of 453 Escherichia coli strains to agglutinate erythrocytes and yeast cells and to attach to human urinary tract epithelial cells was tested. The strains were isolated from the urine of patients with acute pyelonephritis, acute cystitis, or asymptomatic bacteriuria and from the stools of healthy school children. Three main patterns of hemagglutination were found: (i) mannoseresistant agglutination of human erythrocytes alone or simultaneously with mannose-sensitive agglutination of guinea pig erythrocytes: (ii) only mannosesensitive agglutination of guinea pig and other erythrocytes; and (iii) no agglutination. Strains with mannose-resistant agglutination of human erythrocytes alone or in combination with mannose-sensitive hemagglutination attached in high numbers to human urinary tract epithelial cells. Bacteria inducing only mannosesensitive hemagglutination attached in low numbers, and non-agglutinating strains did not bind to the urinary tract epithelial cells. The bacterial surface antigen(s) mediating mannose-resistant hemagglutination of human erythrocytes and attachment to human urinary tract epithelial cells may be one factor selecting for E. coli from among the fecal flora which infect the urinary tract. The highest proportion of strains with this property was found among acute pyelonephritis isolates (77%), and the lowest proportion of strains with this property was found among normal fecal E. coli (16%).