In vitro assays for clinical isolates of sequence type 131 Escherichia coli do not recapitulate in vivo infectivity using a murine model of urinary tract infection.

Abstract

Sequence type 131 isolates are a major cause of cystitis and pyelonephritis. Many studies rely solely on in vitro assays to screen for bacterial virulence factors associated with the pathogenicity of clinical isolates of Escherichia coli. Few studies have compared in vitro findings with in vivo infectivity of clinical isolates. The purpose of this study was to evaluate the correlation between in vitro assays with the ability to cause cystitis and pyelonephritis in a murine model of urinary tract infection. In vitro assays were conducted according to the published protocols and included motility assays, biofilm formation, epithelial cell adhesion and invasion, and curli production. Twenty-one uropathogenic E. coli (UPEC) isolates of E. coli ST131 and non-ST131 were used for both in vivo and in vitro studies. Six mice per isolate were inoculated via urethral catheterization. Colony forming units (CFUs) were determined from bladder and kidneys. In vitro and in vivo correlations were evaluated by multiple linear regression analysis. Pairwise linear regressions showed trendlines with weak positive correlations for motility, adhesion, and invasion and weak negative correlations for hemagglutination, biofilm, and curli production. The ability of E. coli ST131 and non-ST131 clinical isolates to cause cystitis and pyelonephritis varied among strains. The R² Pearson correlation value was less than ±0.5 for any pair, indicating little to no statistical association between in vitro and in vivo findings. These data show that in vitro data are not predictive of the ability of ST131 E. coli to infect and/or cause disease in a mouse model.

Importance: Urinary tract infections (UTIs) affect 150 million people annually, and E. coli ST131, a pandemic clone, has become responsible for a significant portion of those UTIs. How ST131 E. coli has become such a successful strain remains to be elucidated. When evaluating bacterial pathogenicity, it is customary to use in vitro assays to predict isolate virulence and fitness due to lower cost and ease of experimentation compared with in vivo models. It is common to use model organisms like pathogenic E. coli CFT073 or a non-pathogenic K12 lab strain as representatives for the entire species. However, our research has shown that model organisms differ from ST131 E. coli, and in vitro assays are poor predictors of ST131 isolate infectivity in a murine model of UTI. As such, research into the mechanisms of fitness/pathogenesis for ST131 infectivity needs to focus on these organisms rather than other types of UPEC.