Limits...
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.

Show MeSH

Related in: MedlinePlus

Binding of C. jejuni 11168 to uncapped terminal Gal structures.Fluorescence intensities associated with C. jejuni 11168-GS (A) and C. jejuni 11168-O (B) binding to uncapped Gal structures (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.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2654152&req=5

pone-0004927-g001: Binding of C. jejuni 11168 to uncapped terminal Gal structures.Fluorescence intensities associated with C. jejuni 11168-GS (A) and C. jejuni 11168-O (B) binding to uncapped Gal structures (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-GS maintained at 25°C (11168-GS 25°C) exhibited little or no binding to terminal Gal-containing structures present on the array. Of particular note, no binding to the core O-glycan Tn antigen (Galβ1-3GalNAcα1-O-Ser; 1M) or glycans terminated with Galα1-3/4Gal (1K, 1N, 1O, 1P and 2A; see table 2 for structures) was observed (Figure 1A). The growth of 11168-GS at 37°C under a microaerobic atmosphere, which mimics mammalian host-like conditions (11168-GS 37°C), and at 42°C under a microaerobic atmosphere, which mimics avian host-like conditions (11168-GS 42°C), resulted in significantly greater binding (P<0.05) to terminal Gal structures in comparison to 11168-GS 25°C. In fact, C. jejuni 11168-GS 37°C and 11168-GS 42°C bound all terminal Gal structures present on the array (Figure 1A). However, structures such as Galβ1-3GlcNAc, Galβ1-3GalNAc, Galβ1-3GalNAcβ1-4Galβ1-4Glc and Galα1-3Gal (1A, E, F and N respectively; Figure 1 and Table 2) were bound significantly less by 11168-GS 42°C in comparison to 11168-GS 37°C. Conversely, the structure Galβ1-4GlcNAcβ1-3Galβ1-4Glc (1H, Figure 1 and Table 2) was bound significantly more by 11168-GS 42°C than 11168-GS 37°C. Even though some differences in the level of binding to various terminal Gal structures by C. jejuni 11168-GS grown at conditions that mimic mammalian and avian hosts were noted, there appeared to be no significant specificity requirement for a particular glycosidic linkage or underlying sugar.


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 uncapped terminal Gal structures.Fluorescence intensities associated with C. jejuni 11168-GS (A) and C. jejuni 11168-O (B) binding to uncapped Gal structures (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.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0004927-g001: Binding of C. jejuni 11168 to uncapped terminal Gal structures.Fluorescence intensities associated with C. jejuni 11168-GS (A) and C. jejuni 11168-O (B) binding to uncapped Gal structures (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-GS maintained at 25°C (11168-GS 25°C) exhibited little or no binding to terminal Gal-containing structures present on the array. Of particular note, no binding to the core O-glycan Tn antigen (Galβ1-3GalNAcα1-O-Ser; 1M) or glycans terminated with Galα1-3/4Gal (1K, 1N, 1O, 1P and 2A; see table 2 for structures) was observed (Figure 1A). The growth of 11168-GS at 37°C under a microaerobic atmosphere, which mimics mammalian host-like conditions (11168-GS 37°C), and at 42°C under a microaerobic atmosphere, which mimics avian host-like conditions (11168-GS 42°C), resulted in significantly greater binding (P<0.05) to terminal Gal structures in comparison to 11168-GS 25°C. In fact, C. jejuni 11168-GS 37°C and 11168-GS 42°C bound all terminal Gal structures present on the array (Figure 1A). However, structures such as Galβ1-3GlcNAc, Galβ1-3GalNAc, Galβ1-3GalNAcβ1-4Galβ1-4Glc and Galα1-3Gal (1A, E, F and N respectively; Figure 1 and Table 2) were bound significantly less by 11168-GS 42°C in comparison to 11168-GS 37°C. Conversely, the structure Galβ1-4GlcNAcβ1-3Galβ1-4Glc (1H, Figure 1 and Table 2) was bound significantly more by 11168-GS 42°C than 11168-GS 37°C. Even though some differences in the level of binding to various terminal Gal structures by C. jejuni 11168-GS grown at conditions that mimic mammalian and avian hosts were noted, there appeared to be no significant specificity requirement for a particular glycosidic linkage or underlying sugar.

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