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

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

Glycan structures present on array.

<p>Glycan structures present on array.</p

Binding of <i>C. jejuni</i> 11168 to fucosylated structures.

<p>Fluorescence intensities associated with <i>C. jejuni</i> 11168-GS (A) and <i>C. jejuni</i> 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 <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0004927#pone-0004927-t002" target="_blank">Table 2</a>. * Significantly different to 25°C, P<0.05; # Significant difference between 37°C and 42°C, P<0.05.</p

Relative adherence of <i>C. jejuni</i> 11168 to sialidase treated Caco-2 cells.

<p>Relative adherence of 11168-GS (left) and 11168-O (right) to sialidase treated Caco-2 cells with respect to normal adherence levels (25°C, black bar; 37°C, grey bar; and 42°C, white bar).</p

Binding of <i>C. jejuni</i> 11168 to Man containing structures.

<p>Fluorescence intensities associated with <i>C. jejuni</i> 11168-GS (A) and <i>C. jejuni</i> 11168-O (B) binding to Man containing glycans (25°C, black bar; 37°C, grey bar; and 42°C, white bar). For the structure of the individual glycans refer to <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0004927#pone-0004927-t002" target="_blank">Table 2</a>. * Significantly different to 25°C, P<0.05; # Significant difference between 37°C and 42°C, P<0.05.</p

Relative binding of <i>C. jejuni</i> 11168 to Neu5Ac containing structures in the presence of sialyltransferase and sialidase inhibitors.

<p>Relative fluorescent levels of treated 11168-GS 42°C (A) and <i>C. jejuni</i> 11168-O 25°C (B) compared to normal <i>C. jejuni</i> binding to Neu5Ac containing glycans. (Neu5Ac2en, black bar; 4-guanidino-Neu5Ac2en, grey bar; and CMP, white bar). For the structure of the individual glycans refer to <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0004927#pone-0004927-t002" target="_blank">Table 2</a>.</p

Binding of <i>C. jejuni</i> 11168 to Neu5Ac containing structures.

<p>Fluorescence intensities associated with <i>C. jejuni</i> 11168-GS (A) and <i>C. jejuni</i> 11168-O (B) binding to Neu5Ac containing glycans (25°C, black bar; 37°C, grey bar; and 42°C, white bar). For the structure of the individual glycans refer to <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0004927#pone-0004927-t002" target="_blank">Table 2</a>. * Significantly different to 25°C, P<0.05; # Significant difference between 37°C and 42°C, P<0.05.</p

Lectin inhibition of <i>C. jejuni</i> adherence to intestinal cell line.

<p><i>C. jejuni</i> 11168-GS (A) and 11168-O (B) adherence to Caco-2 cells pre-treated with ECA, UEA-1, ConA, LFA, MAA and SNA (25°C, black bar; 37°C, grey bar; and 42°C, white bar). * Significant difference to non-lectin treated control, P<0.05; ND, Not determined.</p