Characterization and Functional Activity of Murine Monoclonal Antibodies Specific for α1,6-Glucan Chain of Helicobacter pylori Lipopolysaccharide

Harrison, Blair A.; Fernández, Heriberto; Chandan, Vandana; Schuster, Myra Wilson; Rademacher, Laura Otth; Toledo, Claudio; Li, Jianjun; Altman, Eleonora

doi:10.1111/j.1523-5378.2011.00860.x

Cited by 12 publications

(11 citation statements)

References 16 publications

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“…For example, the HP0826 mutant in SS1 strain, which is devoid of Lewis antigen, is still able to colonise the host, although less efficiently compared to the wild-type [16,35]. The DD-heptan and the glucan of the O-antigen are not essential for colonisation [19,33,47], while the HP0479 mutant truncated at the Hep residue of the Trio resulted in loss of colonisation ability [22]. The variability of the O-antigen beyond the conserved Trio may enable H .…”

Section: Discussionmentioning

confidence: 99%

The redefinition of Helicobacter pylori lipopolysaccharide O-antigen and core-oligosaccharide domains

Yang

Liao

et al. 2017

PLoS Pathog

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Helicobacter pylori lipopolysaccharide promotes chronic gastric colonisation through O-antigen host mimicry and resistance to mucosal antimicrobial peptides mediated primarily by modifications of the lipid A. The structural organisation of the core and O-antigen domains of H. pylori lipopolysaccharide remains unclear, as the O-antigen attachment site has still to be identified experimentally. Here, structural investigations of lipopolysaccharides purified from two wild-type strains and the O-antigen ligase mutant revealed that the H. pylori core-oligosaccharide domain is a short conserved hexasaccharide (Glc-Gal-DD-Hep-LD-Hep-LD-Hep-KDO) decorated with the O-antigen domain encompassing a conserved trisaccharide (-DD-Hep-Fuc-GlcNAc-) and variable glucan, heptan and Lewis antigens. Furthermore, the putative heptosyltransferase HP1284 was found to be required for the transfer of the third heptose residue to the core-oligosaccharide. Interestingly, mutation of HP1284 did not affect the ligation of the O-antigen and resulted in the attachment of the O-antigen onto an incomplete core-oligosaccharide missing the third heptose and the adjoining Glc-Gal residues. Mutants deficient in either HP1284 or O-antigen ligase displayed a moderate increase in susceptibility to polymyxin B but were unable to colonise the mouse gastric mucosa. Finally, mapping mutagenesis and colonisation data of previous studies onto the redefined organisation of H. pylori lipopolysaccharide revealed that only the conserved motifs were essential for colonisation. In conclusion, H. pylori lipopolysaccharide is missing the canonical inner and outer core organisation. Instead it displays a short core and a longer O-antigen encompassing residues previously assigned as the outer core domain. The redefinition of H. pylori lipopolysaccharide domains warrants future studies to dissect the role of each domain in host-pathogen interactions. Also enzymes involved in the assembly of the conserved core structure, such as HP1284, could be attractive targets for the design of new therapeutic agents for managing persistent H. pylori infection causing peptic ulcers and gastric cancer.

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Section: Discussionmentioning

confidence: 99%

The redefinition of Helicobacter pylori lipopolysaccharide O-antigen and core-oligosaccharide domains

Yang

Liao

et al. 2017

PLoS Pathog

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“…We recently reported that 44.7 % of Chilean H. pylori isolates expressed a1,6-glucan, a polymeric component of the outer core region of LPS (Harrison et al, 2011). Co-expression of a1,6-glucan with Le x and/or Le y antigens in the majority of H. pylori strains examined suggests the importance of this LPS component in the pathogenesis of H. pylori.…”

Section: Discussionmentioning

confidence: 99%

“…The wells were then fixed with methanol and blocked with 200 ml Milk Diluent/Blocking solution (MDB) (KPL) for 2 h at 37 uC. Subsequently, the wells were incubated for 2 h at 37 uC with either 100 ml anti-Le a , -Le b , -Le x or -Le y mAb solution (Signet Laboratories) diluted 1 : 200 in MDB, 100 ml 1C4F9 ascites (anti-a1,6-glucan mAb) (Harrison et al, 2011) diluted 1 : 500 in MDB or rabbit anti-CagA antibody (Austral Biologicals) diluted 1 : 100 in MDB. This was followed by incubation with horseradish peroxidase (HRP)-conjugated anti-mouse IgG+IgM (Caltag) diluted 1 : 1000 in MDB for 1 h at room temperature.…”

Section: Methodsmentioning

confidence: 99%

“…2010). The isolates were screened by whole-cell indirect ELISA (WCE) using commercially available anti-Le monoclonal antibodies (mAbs) and anti-a1,6-glucanspecific mAbs developed in our laboratory (Harrison et al, 2011). Selected strains were further characterized in terms of carbohydrate content by using SDS-PAGE electrophoresis, immunoblotting and chemical and mass spectrometric analyses.…”

Section: Introductionmentioning

confidence: 99%

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Helicobacter pylori isolates from Greek children express type 2 and type 1 Lewis and α1,6-glucan antigens in conjunction with a functional type IV secretion system

Altman

Chandan

Harrison

et al. 2012

Journal of Medical Microbiology

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“…LPS Outer Core Structures in Other H. pylori strains. H. pylori strain J99 outer core is based on reference ; strains 11637(O:1)/P466/H428/H507/UA915/UA948/UA955/J223/CA2/CA4/CA5/CA6/F‐58C/F‐15A/R‐58A/F‐58C/GU2/R‐7A/H607 are based on and n = 2–3; strain PJ1 is based on reference ; PJ2 is based on references , O‐antigen is devoid and glucan: n = 3–9; strains O:6 and MO19 are based on references and m = 5, n (heptan) = 2, and n (glucan) = 2–3. For simplicity, lipid A, the inner core, and the O ‐antigen are not indicated.…”

Section: Structure and Biological Roles Of The Core Oligosaccharidementioning

confidence: 99%

Lipopolysaccharide Structure and Biosynthesis in Helicobacter pylori

Liao

Debowski

et al. 2016

Helicobacter

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This review covers the current knowledge and gaps in Helicobacter pylori lipopolysaccharide (LPS) structure and biosynthesis. H. pylori is a Gram-negative bacterium which colonizes the luminal surface of the human gastric epithelium. Both a constitutive alteration of the lipid A preventing TLR4 elicitation and host mimicry of the Lewis antigen decorated O-antigen of H. pylori LPS promote immune escape and chronic infection. To date, the complete structure of H. pylori LPS is not available, and the proposed model is a linear arrangement composed of the inner core defined as the hexa-saccharide (Kdo-LD-Hep-LD-Hep-DD-Hep-Gal-Glc), the outer core composed of a conserved trisaccharide (-GlcNAc-Fuc-DD-Hep-) linked to the third heptose of the inner core, the glucan, the heptan and a variable O-antigen, generally consisting of a poly-LacNAc decorated with Lewis antigens. Although the glycosyltransferases (GTs) responsible for the biosynthesis of the H. pylori O-antigen chains have been identified and characterized, there are many gaps in regard to the biosynthesis of the core LPS. These limitations warrant additional mutagenesis and structural studies to obtain the complete LPS structure and corresponding biosynthetic pathway of this important gastric bacterium.

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Characterization and Functional Activity of Murine Monoclonal Antibodies Specific for α1,6-Glucan Chain of Helicobacter pylori Lipopolysaccharide

Abstract: Anti-α1,6-glucan mAbs could have potential application in typing and surveillance of H. pylori isolates as well as offer insights into structural requirements for the development of LPS-based vaccine against H. pylori infections.

Cited by 12 publications

References 16 publications

The redefinition of Helicobacter pylori lipopolysaccharide O-antigen and core-oligosaccharide domains

The redefinition of Helicobacter pylori lipopolysaccharide O-antigen and core-oligosaccharide domains

Helicobacter pylori isolates from Greek children express type 2 and type 1 Lewis and α1,6-glucan antigens in conjunction with a functional type IV secretion system

Lipopolysaccharide Structure and Biosynthesis in Helicobacter pylori

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