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Differential carbohydrate recognition by Campylobacter jejuni strain 11168: influences of temperature and growth conditions.

Day CJ, Tiralongo J, Hartnell RD, Logue CA, Wilson JC, von Itzstein M, Korolik V - PLoS ONE (2009)

Bottom Line: 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.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.

View Article: PubMed Central - PubMed

Affiliation: Institute for Glycomics, Griffith University Gold Coast Campus, Queensland, Australia.

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 degrees 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.

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Binding of C. jejuni 11168 to fucosylated structures.Fluorescence intensities associated with C. jejuni 11168-GS (A) and C. jejuni 11168-O (B) binding to fucosylated glycans (25°C, black bar; 37°C, grey bar; and 42°C, white bar). For the structure of the individual glycans refer to Table 2. * Significantly different to 25°C, P<0.05; # Significant difference between 37°C and 42°C, P<0.05.
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pone-0004927-g003: Binding of C. jejuni 11168 to fucosylated structures.Fluorescence intensities associated with C. jejuni 11168-GS (A) and C. jejuni 11168-O (B) binding to fucosylated glycans (25°C, black bar; 37°C, grey bar; and 42°C, white bar). For the structure of the individual glycans refer to Table 2. * Significantly different to 25°C, P<0.05; # Significant difference between 37°C and 42°C, P<0.05.

Mentions: C. jejuni 11168 showed significant binding to a range of fucosylated glycans. C. jejuni 11168-GS maintained at 25°C, bound only approximately 50% of the fucosylated glycans present on the array (Figure 3A). Of particular note was the fact that no binding was observed for 11168-GS 25°C to the trisaccharides Lewisa (Galβ1-3(Fucα1-4)GlcNAc; Lea; 7J) and Lewisx (Galβ1-4(Fucα1-3)GlcNAc; Lex; 7I) (Figure 3A), even though detectable binding to other glycans terminated with Lea and Lex was observed (e.g. 7B and 7C, respectively). Further analysis found that 11168-GS 25°C bound monofucosylated glycans with a disaccharide backbone (7F, 7G, 7H, 7I, 7J, 7K, 7M, 7O, 8A and 8B) to a lesser extent than monofucosylated tetrasaccharide or larger glycans (7A, 7B, 7C, 8C and 8D; P = 0.04). As previously observed for 11168-GS binding to terminal Gal (Figure 1A), 11168-GS binding to Fuc was also dependent on the growth/maintenance conditions. That is, fucosylated structures were bound by 11168-GS 37°C and 11168-GS 42°C to a significantly greater extent than 11168-GS 25°C (Figure 3A).


Differential carbohydrate recognition by Campylobacter jejuni strain 11168: influences of temperature and growth conditions.

Day CJ, Tiralongo J, Hartnell RD, Logue CA, Wilson JC, von Itzstein M, Korolik V - PLoS ONE (2009)

Binding of C. jejuni 11168 to fucosylated structures.Fluorescence intensities associated with C. jejuni 11168-GS (A) and C. jejuni 11168-O (B) binding to fucosylated glycans (25°C, black bar; 37°C, grey bar; and 42°C, white bar). For the structure of the individual glycans refer to Table 2. * Significantly different to 25°C, P<0.05; # Significant difference between 37°C and 42°C, P<0.05.
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Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2654152&req=5

pone-0004927-g003: Binding of C. jejuni 11168 to fucosylated structures.Fluorescence intensities associated with C. jejuni 11168-GS (A) and C. jejuni 11168-O (B) binding to fucosylated glycans (25°C, black bar; 37°C, grey bar; and 42°C, white bar). For the structure of the individual glycans refer to Table 2. * Significantly different to 25°C, P<0.05; # Significant difference between 37°C and 42°C, P<0.05.
Mentions: C. jejuni 11168 showed significant binding to a range of fucosylated glycans. C. jejuni 11168-GS maintained at 25°C, bound only approximately 50% of the fucosylated glycans present on the array (Figure 3A). Of particular note was the fact that no binding was observed for 11168-GS 25°C to the trisaccharides Lewisa (Galβ1-3(Fucα1-4)GlcNAc; Lea; 7J) and Lewisx (Galβ1-4(Fucα1-3)GlcNAc; Lex; 7I) (Figure 3A), even though detectable binding to other glycans terminated with Lea and Lex was observed (e.g. 7B and 7C, respectively). Further analysis found that 11168-GS 25°C bound monofucosylated glycans with a disaccharide backbone (7F, 7G, 7H, 7I, 7J, 7K, 7M, 7O, 8A and 8B) to a lesser extent than monofucosylated tetrasaccharide or larger glycans (7A, 7B, 7C, 8C and 8D; P = 0.04). As previously observed for 11168-GS binding to terminal Gal (Figure 1A), 11168-GS binding to Fuc was also dependent on the growth/maintenance conditions. That is, fucosylated structures were bound by 11168-GS 37°C and 11168-GS 42°C to a significantly greater extent than 11168-GS 25°C (Figure 3A).

Bottom Line: 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.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.

View Article: PubMed Central - PubMed

Affiliation: Institute for Glycomics, Griffith University Gold Coast Campus, Queensland, Australia.

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 degrees 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.

Show MeSH
Related in: MedlinePlus