Recognition of Lewis x Derivatives Present on Mucins by Flagellar Components of
            <i>Pseudomonas aeruginosa</i>

Scharfman, A; Arora, Shiwani K.; Delmotte, Philippe; Brussel, Edwige Van; Mazurier, Joël; Ramphal, Reuben; Roussel, Philippe

doi:10.1128/iai.69.9.5243-5248.2001

Cited by 89 publications

(62 citation statements)

References 27 publications

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“…Three of four serotype O1 and all three O11 isolates were flagellin type a. The overall type a:type b ratio (58 %:41 %) suggested a predominance of flagellin type a, but not to the same degree as the keratitis isolates in this study (76 %:24 % There is evidence not only that P. aeruginosa flagella play a role in mucin binding, but that such binding varies depending on flagellar type (Scharfman et al, 2001). It is possible, therefore, that the predominance of a flagellin, and hence a flagellar, type associated with a particular site of infection may be influenced by mucin-binding specificities.…”

Section: Discussionmentioning

confidence: 43%

Genotypic and phenotypic characteristics of Pseudomonas aeruginosa isolates associated with ulcerative keratitis

Winstanley

Kaye

Neal

et al. 2005

Journal of Medical Microbiology

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A collection of 63 isolates of Pseudomonas aeruginosa associated with ulcerative keratitis, collected from six centres in England, were typed using serotyping and random amplified polymorphic DNA-PCR, and screened for several variable virulence-related genotypes and phenotypes. Sixty-one percent of the isolates were of either serotype O1 or serotype O11, but there was no evidence for a common clone. The majority of isolates (59 %) were PCR-positive for exoU rather than for exoS (38 %), and carried a-type fliC genes (76 %) rather than b-type (24 %). Isolates were PCR-positive for pyoverdine-receptor types at a prevalence of 38 % for type I, 46 % for type II and 8 % for type III. All but one of the isolates exhibited twitching activity. There was a correlation between the presence of exoS and twitching activity (P ¼ 0 . 04), suggesting that a combination of exoS genotype and good twitching activity may have a role to play in ExoU-independent corneal virulence. INTRODUCTIONPseudomonas aeruginosa, a major opportunistic pathogen, is a common cause of bacterial keratitis, especially in contact lens wearers. Infection with P. aeruginosa leads to a rapid destructive keratitis and potential perforation within 2-4 days (Fleiszig & Evans, 2002). Generally P. aeruginosa-related keratitis tends to have a poor outcome in terms of corneal scarring and response to treatment. The pathogenesis of P. aeruginosa keratitis consists of colonization of the cornea, induction of proinflammatory cytokines (TNF-AE, IL-1) (Kernacki et al., 1998;Rudner et al., 2000), the migration of neutrophils into the cornea to kill and remove the pathogen (Pillar & Hobden, 2002), and subsequent corneal damage due to the neutrophil response and bacterial factors.Flagella and pili play potential roles in the pathogenicity of P. aeruginosa as adhesins (Feldman et al., 1998; Comolli et al., 1999). Type IV pili are surface appendages with a role in adherence (Hahn, 1997) or non-flagellar 'twitching' motility that are produced by many bacterial pathogens. It has been demonstrated that mutants of P. aeruginosa defective in twitching motility show reduced ability to colonize the cornea in a mouse model (Zolfaghar et al., 2003). Thus, twitching motility appears to contribute to the role of pili in corneal infection. In addition, it has been shown that flagellum assembly mutants are attenuated in their ability to invade corneal epithelial cells (Fleiszig et al., 2001). The flagella of P. aeruginosa also contribute to the inflammatory responses of corneal epithelial cells in a TLR5-NF-kB signalling pathway-dependent manner (Zhang et al., 2003). P. aeruginosa is capable of secreting a number of proteins with potential roles in pathogenicity. These include proteases (alkaline protease, staphylolytic protease, elastase, protease IV), heat-labile and heat-stable haemolysins, phospholipase C and exotoxins A, S, T, U and Y (Kawaharajo et al., 1974;Kessler et al., 1977;Ohman et al., 1980;Pillar & Hobden, 2002;Sato et al., 2003;Schulert et al., 2003). Clinical isolates o...

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Section: Discussionmentioning

confidence: 43%

Genotypic and phenotypic characteristics of Pseudomonas aeruginosa isolates associated with ulcerative keratitis

Winstanley

Kaye

Neal

et al. 2005

Journal of Medical Microbiology

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“…Previous studies have suggested that glycosphingolipids may serve as AP receptors for type IV pili (10,47), although these findings remain controversial (13). Flagellar components have been shown to bind to Lewis X derivatives that are found on secreted mucins (48). Moreover, Toll-like receptor 5, predominantly found on the AP surface, has been found to bind flagellin (24).…”

Section: Discussionmentioning

confidence: 99%

Pseudomonas aeruginosa -Mediated Damage Requires Distinct Receptors at the Apical and Basolateral Surfaces of the Polarized Epithelium

2010

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Pseudomonas aeruginosa, an important opportunistic pathogen of humans, exploits epithelial damage to establish infection. We have rigorously explored the role of N-glycoproteins and heparan sulfate proteoglycans (HSPGs) in P. aeruginosa-mediated attachment and subsequent downstream events at the apical (AP) and basolateral (BL) surfaces of polarized epithelium. We demonstrate that the N-glycan chains at the AP surface are necessary and sufficient for binding, invasion, and cytotoxicity to kidney (MDCK) and airway (Calu-3) cells grown at various states of polarization on Transwell filters. Upregulation of N-glycosylation enhanced binding, whereas pharmacologic inhibition of N-glycosylation or infection of MDCK cells defective in N-glycosylation resulted in decreased binding. In contrast, at the BL surface, the HS moiety of HSPGs mediated P. aeruginosa binding, cytotoxicity, and invasion. In incompletely polarized epithelium, HSPG abundance was increased at the AP surface, explaining its increased susceptibility to P. aeruginosa colonization and damage. Using MDCK cells grown as three-dimensional cysts as a model for epithelial organs, we show that P. aeruginosa specifically colocalized with HS-rich areas at the BL membrane but with complex N-glycans at the AP surface. Finally, P. aeruginosa bound to HS chains and N-glycans coated on plastic surfaces, showing the highest binding affinity toward isolated HS chains. Together, these findings demonstrate that P. aeruginosa recognizes distinct receptors on the AP and BL surfaces of polarized epithelium. Changes in the composition of N-glycan chains and/or in the distribution of HSPGs may explain the enhanced susceptibility of damaged epithelium to P. aeruginosa.Ninety-five percent of all infectious agents enter through mucosal surfaces of the gastrointestinal, genitourinary, and respiratory tracts (reviewed in reference 35). These mucosal surfaces are usually lined by a single layer of epithelial cells, which serves as the primary barrier against the entry of most infectious agents and can be considered a primary component of the innate immune system. Epithelial cells form highly polarized cell layers with apical (AP) and basolateral (BL) surfaces that exhibit distinct protein, lipid, and glycoconjugate compositions. Pseudomonas aeruginosa is a ubiquitous opportunistic pathogen of humans that exploits injured mucosa to cause acute and chronic infections with high morbidity and mortality (reviewed in references 26 and 31). In the setting of epithelial injury and immunocompromise, this Gram-negative pathogen causes serious infections in patients with extensive burns, corneal trauma, or catheter-related bladder injury or in those on ventilators. In addition, P. aeruginosa chronically colonizes the lungs of patients with cystic fibrosis (CF) (4), leading to severe pulmonary damage and death. Despite aggressive antibiotic therapy, the fatality rate for many P. aeruginosa infections is 40%, and new approaches to treatment are even more critical now that antibiotic resistance...

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“…FliD also functions as a cap that prevents FliC subunits from diffusing out of the cell. Additionally, in Pseudomonas aeruginosa and Clostridium difficile, FliD has been reported to function as a mucin-specific adhesin (18,24).…”

Section: Discussionmentioning

confidence: 99%

Simultaneous Display of Multiple Foreign Peptides in the FliD Capping and FliC Filament Proteins of the Escherichia coli Flagellum

Majander

Korhonen

Westerlund-Wikström

2005

Appl Environ Microbiol

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The bacterial flagellum is composed of more than 20 different proteins. The filament, which constitutes the major extracellular part of the flagellum, is built up of approximately 20,000 FliC molecules that assemble at the growing distal end of the filament. A capping structure composed of five FliD molecules located at the tip of the filament promotes polymerization of FliC. Lack of FliD leads to release of the subunits into the growth medium. We show here that FliD can be successfully used in bacterial surface display. We tested various insertion sites in the capping protein, and the optimal region for display was at the variable region in FliD. Deletion and/or insertion at other sites resulted in decreased formation of flagella. We further developed the technique into a multihybrid display system in which three foreign peptides are simultaneously expressed within the same flagellum, i.e., D repeats of FnBPA from Staphylococcus aureus at the tip and fragments of YadA from Yersinia enterocolitica as well as SlpA from Lactobacillus crispatus along the filament. This technology can have biotechnological applications, e.g., in simultaneous delivery of several effector molecules.The flagellum is the bacterial motility organelle and is composed of more than 20 different proteins that build up the basal body, the hook, the filament, and the flagellum-specific type III secretion system. The organelle traverses the bacterial inner and outer membranes and forms a 2-nm-wide central channel, inside which the FliC flagellin subunits are transported to the growing distal end, where they assemble to a filament of approximately 20,000 FliC copies (1). Flagellar assembly is strictly regulated by hierarchical expression of the genes encoding flagellar components and proceeds in a highly ordered fashion from the basal body components and the secretion machinery towards the hook and the filament (11).The capping protein FliD (also called HAP2) has a central role in flagellar polymerization. After completion of the synthesis of hook-basal body complex, the FliD cap is attached to the tip of the hook and newly exported FliC monomers insert between the hook and the cap. The cap is essential for polymerization of FliC monomers as well as assembly and growth of the flagellar filament. Deletion of fliD leads to inability of transported FliC to polymerize into filaments and to their secretion into the growth medium (6,8). A high-resolution map based on electron microscopy revealed that five FliD molecules assemble to form the stool-like flagellar cap that consists of five leg-like domains and a plate-like structure (Fig.

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Recognition of Lewis x Derivatives Present on Mucins by Flagellar Components of Pseudomonas aeruginosa

Cited by 89 publications

References 27 publications

Genotypic and phenotypic characteristics of Pseudomonas aeruginosa isolates associated with ulcerative keratitis

Genotypic and phenotypic characteristics of Pseudomonas aeruginosa isolates associated with ulcerative keratitis

Pseudomonas aeruginosa -Mediated Damage Requires Distinct Receptors at the Apical and Basolateral Surfaces of the Polarized Epithelium

Simultaneous Display of Multiple Foreign Peptides in the FliD Capping and FliC Filament Proteins of the Escherichia coli Flagellum

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