Highly conserved type 1 pili promote enterotoxigenic E. coli pathogen-host interactions.

Alaullah Sheikh,Scott J Hultgren,Rasheduzzaman Rashu,James M Fleckenstein,F Matthew Kuhlman,Matthew A Ciorba,Yasmin Ara Begum,Firdausi Qadri,Ruifu Yang

doi:10.1371/journal.pntd.0005586

Abstract

Enterotoxigenic Escherichia coli (ETEC), defined by their elaboration of heat-labile (LT) and/or heat-stable (ST) enterotoxins, are a common cause of diarrheal illness in developing countries. Efficient delivery of these toxins requires ETEC to engage target host enterocytes. This engagement is accomplished using a variety of pathovar-specific and conserved E. coli adhesin molecules as well as plasmid encoded colonization factors. Some of these adhesins undergo significant transcriptional modulation as ETEC encounter intestinal epithelia, perhaps suggesting that they cooperatively facilitate interaction with the host. Among genes significantly upregulated on cell contact are those encoding type 1 pili. We therefore investigated the role played by these pili in facilitating ETEC adhesion, and toxin delivery to model intestinal epithelia. We demonstrate that type 1 pili, encoded in the E. coli core genome, play an essential role in ETEC virulence, acting in concert with plasmid-encoded pathovar specific colonization factor (CF) fimbriae to promote optimal bacterial adhesion to cultured intestinal epithelium (CIE) and to epithelial monolayers differentiated from human small intestinal stem cells. Type 1 pili are tipped with the FimH adhesin which recognizes mannose with stereochemical specificity. Thus, enhanced production of highly mannosylated proteins on intestinal epithelia promoted FimH-mediated ETEC adhesion, while conversely, interruption of FimH lectin-epithelial interactions with soluble mannose, anti-FimH antibodies or mutagenesis of fimH effectively blocked ETEC adhesion. Moreover, fimH mutants were significantly impaired in delivery of both heat-stable and heat-labile toxins to the target epithelial cells in vitro, and these mutants were substantially less virulent in rabbit ileal loop assays, a classical model of ETEC pathogenesis. Collectively, our data suggest that these highly conserved pili play an essential role in virulence of these diverse pathogens.

Highlights

Among young children under five years of age in developing countries, diarrhea is a leading cause of morbidity and mortality
Effective engagement of intestinal epithelial cells is essential for Enterotoxigenic Escherichia coli (ETEC) pathogenicity
Pathovar specific plasmid-encoded adhesin structures known as colonization factors (CFs) have been a principal target for vaccines

Summary

Introduction

Among young children under five years of age in developing countries, diarrhea is a leading cause of morbidity and mortality. Enterotoxigenic E. coli (ETEC) is one of the most common causes of moderate to severe diarrheal illness and deaths due to diarrhea in young children and incidentally is the leading bacterial cause of diarrhea [1]. These bacteria are a leading cause of hospitalization due to severe diarrhea in adults in developing countries [2] and are perennially the predominant cause of diarrheal illness among travelers to the endemic regions [3, 4]. These pathogens contribute to a complex pattern of poverty, repeated enteric infections, environmental enteropathy [9], and developmental impairment

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