Differential Carbohydrate Recognition by Campylobacter jejuni Strain 11168: Influences of Temperature and Growth Conditions

Markus M Heimesaat,Christopher J Day,Regan D Hartnell,Mark Von Itzstein,Victoria Korolik,Joe Tiralongo,Jennifer C Wilson,Carie-Anne Logue

doi:10.1371/journal.pone.0004927

Abstract

The pathogenic clinical strain NCTC11168 was the first Campylobacter jejuni strain to be sequenced and has been a widely used laboratory model for studying C. jejuni pathogenesis. However, continuous passaging of C. jejuni NCTC11168 has been shown to dramatically affect its colonisation potential. Glycan array analysis was performed on C. jejuni NCTC11168 using the frequently passaged, non-colonising, genome sequenced (11168-GS) and the infrequently passaged, original, virulent (11168-O) isolates grown or maintained under various conditions. Glycan structures recognised and bound by C. jejuni included terminal mannose, N-acetylneuraminic acid, galactose and fucose. Significantly, it was found that only when challenged with normal oxygen at room temperature did 11168-O consistently bind to sialic acid or terminal mannose structures, while 11168-GS bound these structures regardless of growth/maintenance conditions. Further, binding of un-capped galactose and fucosylated structures was significantly reduced when C. jejuni was maintained at 25°C under atmospheric oxygen conditions. These binding differences identified through glycan array analysis were confirmed by the ability of specific lectins to competitively inhibit the adherence of C. jejuni to a Caco-2 intestinal cell line. Our data suggests that the binding of mannose and/or N-acetylneuraminic acid may provide the initial interactions important for colonisation following environmental exposure.

Highlights

Carbohydrates that modify proteins and lipids play a key role in numerous cell recognition events, including those involved in the regulation of the immune system [1], and in the attachment of pathogenic organisms to host tissue [2]
Proteincarbohydrate interactions have been identified as adherence factors for numerous commensal and pathogenic bacteria including Pseudomonas aeruginosa[3,4,5], Helicobacter pylori [6,7,8,9,10] and Escherichia coli [11]
In this study we demonstrate that glycan array technology represents a powerful tool for the identification and characterisation of novel bacteria-glycoconjugate interactions

Summary

Introduction

Carbohydrates (or glycans) that modify proteins and lipids play a key role in numerous cell recognition events, including those involved in the regulation of the immune system [1], and in the attachment of pathogenic organisms to host tissue [2]. Proteincarbohydrate interactions have been identified as adherence factors for numerous commensal and pathogenic bacteria including Pseudomonas aeruginosa[3,4,5], Helicobacter pylori [6,7,8,9,10] and Escherichia coli [11]. The identification and characterisation of carbohydrate binding proteins (or lectins) has been greatly enhanced through the development of glycan array technology [12]. C. jejuni is a zoonotic pathogen, being a commensal organism in poultry and other wildlife. C. jejuni is a thermophilic organism, requiring temperatures in the range of 32uC to 45uC for growth, but it can survive at lower temperatures in the environment [18]

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