J Med Microbiol International Journal of Systematic and Evolutionary Microbiology
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via CrossRef
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Thomas, R.
Right arrow Articles by Brooks, T.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Thomas, R.
Right arrow Articles by Brooks, T.
Agricola
Right arrow Articles by Thomas, R.
Right arrow Articles by Brooks, T.
J Med Microbiol 53 (2004), 833-840; DOI: 10.1099/jmm.0.45643-0
© 2004 Society for General Microbiology
ISSN 0022-2615

Common oligosaccharide moieties inhibit the adherence of typical and atypical respiratory pathogens

Richard Thomas1 and Tim Brooks2

1Defence Science & Technology Laboratories (Dstl), Biomedical Sciences, Porton Down, Salisbury, Wiltshire SP4 0JQ, UK 2Health Protection Agency (HPA), Porton Down, Salisbury, Wiltshire SP4 0JG, UK

Correspondence Richard Thomas rjthomas{at}dstl.gov.uk

Received February 25, 2004
Accepted April 29, 2004

Intervention in bacterial adhesion to host cells is a novel method of overcoming current problems associated with antibiotic resistance. Antibiotic-resistant strains of bacteria that cause respiratory tract infections are a problem in hospitals and could be used in bioterrorist attacks. A range of bacterial species was demonstrated to attach to an alveolar epithelial (A549) cell line. In all cases, cell surface oligosaccharides were important in attachment, demonstrated by reduced adhesion when A549 cells were pre-treated with tunicamycin. Bacillus anthracis and Yersinia pestis displayed a restricted tropism for oligosaccharides compared to the environmental, opportunistic pathogens, Pseudomonas aeruginosa, Burkholderia cenocepacia, Burkholderia pseudomallei and Legionella pneumophila. The compound with the greatest anti-adhesion activity was p-nitrophenol. Other generic attachment inhibitors included the polymeric saccharides (dextran and heparin), GalNAcß1-4Gal, GalNAcß1-3Gal, Galß1-4GlcNAc and Galß1-3GlcNAc. Burkholderia pseudomallei attachment was particularly susceptible to oligosaccharide inhibition. Combinations of such compounds may serve as a novel generic therapeutics for respiratory tract infections.


   INTRODUCTION
TOP
 INTRODUCTION
METHODS
 RESULTS AND DISCUSSION
 ACKNOWLEDGEMENTS
 REFERENCES
 
The development of antibiotic resistance amongst pathogenic bacteria is a frequent occurrence and of great concern for public health. This problem is associated with many bacteria that cause community- and hospital-acquired pneumonia such as Streptococcus pneumoniae, Haemophilus influenzae, Pseudomonas aeruginosa and Burkholderia cenocepacia (Lister, 2000; Lynch, 2001; Spencer, 1995; Tan, 2003). Problems with antibiotic resistance can also extend to those bacteria that could be used in bioterrorism. It is likely that such bacterial threat agents would be delivered by the aerosol route resulting in respiratory presentation of the disease (Cieslak & Eitzen, 2000; Leggiadro, 2000). Given the ease of natural acquisition of antibiotic resistance genes and genetic manipulation, it is prudent to research therapeutic alternatives to antibiotics.

Attachment to cell surfaces is one of the key steps in bacterial pathogenesis, often involving oligosaccharides located on the host cell surface and bacterial adhesins (Karlsson et al., 1992; Ofek & Sharon, 1990). The terminal di- or trisaccharide units of these oligosaccharides may be used to inhibit these interactions, preventing attachment and hence disease (Zopf & Roth, 1996). The theory has been proven in vitro with a range of bacterial species. For example, lacto-N-neotetraose and asialoganglioside-GM1 (asialo-GM1) inhibit attachment of S. pneumoniae to alveolar epithelial cells (Tong et al., 1999). Similarly 3-sialyllactose inhibits Helicobacter pylori attachment to gastric epithelial cells (Simon et al., 1997). Asialoganglioside-GM1 and -GM2 are used as receptors by a number of respiratory pathogens, which specifically bind their terminal GalNAcß1-3Gal and GalNAcß1-4Gal moieties (Krivan et al., 1988a, b, 1991). The principle has also been proven to be valid in vivo. The symptoms of pneumococcal pneumonia were alleviated in a rabbit model by administration of lacto-N-neotetraose or its {alpha}2-3- and {alpha}2-6-sialylated derivatives (Idänpään-Heikkilä et al., 1997). Aerosolized dextran significantly reduced murine pneumonia caused by P. aeruginosa (Bryan et al., 1999). Mannose and globotetraose (Gal{alpha}1-4Gal terminal moiety) inhibited Escherichia coli urinary tract infection in BALB/c mice (Edén et al., 1982). This study aimed to determine the potential of oligosaccharides to inhibit the attachment to alveolar epithelial cells of a range of pathogens that can cause pneumonic disease.


   METHODS
TOP
 INTRODUCTION
 METHODS
RESULTS AND DISCUSSION
 ACKNOWLEDGEMENTS
 REFERENCES
 
Materials.

Tissue culture reagents, p-nitrophenol and tunicamycin were purchased from Sigma-Aldrich. Saccharides were purchased as powders from Dextra Laboratories Ltd, Reading, UK or Sigma-Aldrich.

Organisms and growth conditions.

Bacillus cereus ATCC 52522, Yersinia enterocolitica ATCC 35669 and Yersinia pseudotuberculosis ATCC 29833T were obtained from the American Type Culture Collection. P. aeruginosa PAK, Legionella pneumophila CORBY and E. coli O127 : H6 were kind donations from Dr J. Boyd (University of Calgary, Canada), Dr G. Blum-Oehler (Universität Würzburg, Germany) and Dr H. Smith (PHLS, Colindale, UK), respectively. Burkholderia cenocepacia 9091, Burkholderia pseudomallei K96243, Yersinia pestis GB, Salmonella typhimurium LT2 and Bacillus anthracis UM23CL2 (pXO– pXO2–) were from the site culture collection. The bacteria were routinely cultured on agar plates prior to broth inoculation. P. aeruginosa, Burkholderia cenocepacia, Burkholderia pseudomallei, Bacillus cereus and Bacillus anthracis were grown on Luria–Bertani (LB) agar plates at 37 °C for 24 h. L. pneumophila was cultured on buffered charcoal yeast extract (BCYE) medium supplemented with 10 % (v/v) Legionella BCYE supplement (Oxoid) for 72 h at 37 °C in an atmosphere of 5 % (v/v) CO2. Y. pestis was cultured on blood agar base (BAB) at 28 °C for 48 h. To obtain cell densities of 108 c.f.u. ml–1 for the adhesion studies, single colonies were inoculated into broth cultures as described. P. aeruginosa, Burkholderia cenocepacia, Burkholderia pseudomallei, Bacillus cereus, Bacillus anthracis, E. coli, S. typhimurium, Y. enterocolitica and Y. pseudotuberculosis were grown in LB broth for 16 h at 37 °C. L. pneumophila was grown for 24 h at 37 °C in nutrient broth supplemented with 10 % (v/v) Legionella BCYE supplement in an atmosphere of 5 % (v/v) CO2. Y. pestis was grown in BAB broth at 28 °C for 16 h. All broth cultures were incubated statically. Stocks were stored at –80 °C in the respective base broth supplemented with 10 % (v/v) glycerol.

Culture of alveolar type II pneumocytes.

Adherent A549 cells were cultured in tissue culture flasks containing Dulbecco's Modified Eagle's Medium (DMEM) supplemented with 10 % (v/v) fetal calf serum and 2 mM glutamine. Cells were incubated at 37 °C in an atmosphere of 5 % (v/v) CO2 and subcultured 1 : 3 every 3–4 days. For the purpose of adhesion assays, the cells were rinsed twice with PBS before incubation in the presence of trypsin/0.05 % (w/v) EDTA. After removal of the trypsin solution, the cells were resuspended in fresh medium, seeded in 96-well microtitre plates to a density of 50 000 cells per well and incubated overnight as described previously to produce confluent monolayers ready for the adhesion assay. In experiments that required reduced expression of surface oligosaccharides the A549 cells were cultured in the presence of 0.55 µg tunicamycin ml–1 for 96 h prior to completion of the adhesion assay.

Adhesion assay.

Confluent monolayers in 96-well microtitre plates were washed with PBS. Non-specific binding was blocked by incubation for 1 h at 37 °C with 0.5 % (w/v) BSA before rinsing twice with PBS. Bacteria at a density of 107 c.f.u. ml–1 were incubated for 1 h at room temperature in a 1 : 1 ratio with various saccharide concentrations (Table 1). The relevant concentrations were prepared in tissue culture medium. Controls were always performed with no saccharide. A 100 µl volume of the bacteria-saccharide solution was added to the A549 cells and incubated for 1 h at 37 °C. Non-adherent bacteria were removed by rinsing five times with PBS. Cells were lysed by incubation for 30 min at 37 °C with a 0.1 % (v/v) Triton X-100 solution. Serial dilutions were performed in PBS and 100 µl volumes were plated in duplicate on to the organism-specific plates and incubated accordingly. The percentage inhibition was calculated by the equation: (control–test/control) x 100 %.


View this table:
[in this window]
[in a new window]
  		Table 1. Oligosaccharide inhibition of Burkholderia cenocepacia 9091, L. pneumophila CORBY and P. aeruginosa PAK attachment to A549 cells Inoculum of 107 c.f.u. ml–1, results are the means of three independent experiments.
 

Cytotoxicity assay.

The CellTitre 96 non-radioactive cell proliferation assay (Promega) was used for all cytotoxicity assays in accordance with the manufacturer's instructions.

Statistical analysis.

All experiments were performed as three or five independent assays. Assays were performed in triplicate per experimental run. Results are expressed as the mean value and standard deviations (SD) are provided where appropriate. P-values were calculated using paired t-tests.


   RESULTS AND DISCUSSION
TOP
 INTRODUCTION
 METHODS
 RESULTS AND DISCUSSION
ACKNOWLEDGEMENTS
 REFERENCES
 
Attachment of respiratory and enteric pathogens to A549 cells

Initial experiments determined the relative adhesion to A549 cells by bacteria that occupy different anatomical niches (Fig. 1). None of the bacterial species examined demonstrated appreciable cytotoxicity towards A549 cells under the assay conditions (data not shown). In general, the six bacterial species associated with pneumonic infections (Bacillus anthracis, Burkholderia cenocepacia, Burkholderia pseudomallei, P. aeruginosa, L. pneumophila and Y. pestis) adhered more efficiently to A549 cells than the enteric bacteria (P < 0.05). The tropism displayed by the bacteria is indicative of the expression of adhesins required for colonization of particular niches within the body. For example, those bacteria that cause respiratory tract infections (e.g. P. aeruginosa) would be expected to express adhesins specific for ligands located on the surface of the A549 cells. Conversely, bacteria that cause enteric disease (e.g. E. coli) would not express such adhesins (or to a lesser degree) and hence attachment is lower than that displayed by respiratory pathogens. Interestingly, Y. enterocolitica and Y. pseudotuberculosis both adhered to A549 cells as efficiently as L. pneumophila and Bacillus anthracis. This could indicate that the enteric yersiniae express a wide range of adhesins enabling attachment to a broad range of tissues. The relatively low attachment of L. pneumophila compared to other respiratory pathogens could be a result of a preference to invade phagocytic cells, such as amoebae or alveolar macrophages, rather than epithelial cells (Kwaik, 2000).



View larger version (28K):
[in this window]
[in a new window]
  		Fig. 1 Relative attachment of respiratory and enteric pathogens to A549 cells. An inoculum of 107 c.f.u. ml–1 was used. Results are the means of five independent experiments, error bars denote SD.

 

It is interesting that the enteric yersiniae were much less adherent to the alveolar cell line than Y. pestis. This implies the expression of adhesins specific to Y. pestis that may mediate pneumonic disease. Adhesins have been identified in Y. pestis. The pH 6 antigen is expressed at pH 6 and 37 °C, forming fibrils that bind asialo-GM1, asialo-GM2 and lactosylceramide (Lindler & Tall, 1993; Payne et al., 1998). The plasminogen activator is an outer membrane serine protease that has also been shown to bind laminin, heparin and globoseries glycolipids that contain terminal GalNAcß1-3Gal moieties (Kienle et al., 1992; Lähteenmäki et al., 1998). Recently, the Y. pestis genome has been shown to possess two homologues to the Erwinia HecA adhesin (Rojas et al., 2002). It is unknown whether the homologues are expressed and functional. Other possible adhesins include the capsule (F1 antigen) and lipopolysaccharides (LPS) (Straley, 1993).

Oligosaccharides on the surface of A549 cells mediate attachment

Bacteria often attach to the oligosaccharide moiety of glycoproteins and glycolipids located in the host cell membrane (Karlsson et al., 1992; Ofek & Sharon, 1990). Tunicamycin is a specific inhibitor of glycosylation in eukaryotic cells, and can be used to determine the effects of glycosylation on bacterial adhesion (Cundell & Tuomanen, 1994). Tunicamycin inhibited the attachment of each bacterial species examined to A549 cells to varying degrees: Bacillus anthracis, 37.9 ± 5.7 %; Burkholderia cenocepacia, 88.4 ± 6.2 %; Burkholderia pseudomallei, 66.6 ± 1.2 %; L. pneumophila, 83.5 ± 5.1 %; P. aeruginosa, 45.9 ± 9.7 % and Y. pestis, 64.9 ± 4.0 %. The evidence indicates that oligosaccharides located on the A549 cell surface are involved in the attachment of these pathogens. The finding that tunicamycin treatment only inhibits P. aeruginosa attachment by ~46 % is surprising, given the range of adhesins it expresses and saccharide structures it can bind (Tables 1 and 3; Bargouthi et al., 1996; Bryan et al., 1999; Krivan et al., 1988b; Ramphal et al., 1991). Tunicamycin inhibits the biosynthesis of high mannose and complex-type glycoproteins (Cundell & Tuomanen, 1994), however, low level glycosylation could still occur, resulting in attachment of P. aeruginosa to saccharides unaffected by tunicamycin treatment. The low value obtained for Bacillus anthracis is consistent with the evidence that few saccharide structures inhibited attachment to a large degree. Additionally, tunicamycin tends to inhibit the synthesis of high mannose oligosaccharides, and mannose was a poor inhibitor of Bacillus anthracis in comparison to the other bacteria tested (Table 2).


View this table:
[in this window]
[in a new window]
  		Table 3. Bacterial pathogens associated with human lower respiratory disease and oligosaccharide structures they bind ND, Not determined.
 

View this table:
[in this window]
[in a new window]
  		Table 2. Oligosaccharide inhibition of Bacillus anthracis UM23CL2 (pXO1– pXO2–), Burkholderia pseudomallei K96243 and Y. pestis GB attachment to A549 cells Inoculum of 107 c.f.u. ml–1. The saccharide concentrations used are as shown in Table 1. Results are the means of three independent experiments.
 

Oligosaccharide inhibition of respiratory pathogen binding

A panel of oligosaccharides was screened for their ability to inhibit the attachment of six bacterial species associated with respiratory infection (Bacillus anthracis, Burkholderia cenocepacia, Burkholderia pseudomallei, L. pneumophila, P. aeruginosa and Y. pestis; Tables 1 and 2). Incubation with oligosaccharides did not adversely affect A549 cells as indicated by the absence of any cytotoxic effects and microscopic examination (data not shown). The range of saccharide structures that inhibited the attachment of certain bacterial species was surprising and probably reflected the fact that environmental opportunistic pathogens (P. aeruginosa, Burkholderia cenocepacia, Burkholderia pseudomallei and L. pneumophila) would require a range of adhesins to allow attachment to environmental surfaces. In contrast, the more specialized pathogens (Bacillus anthracis and Y. pestis) bind a narrower range of saccharides due to the reduced requirement for attachment to environmental substrata.

Large polymeric saccharides such as dextran, dextran sulphate and heparin were particularly adept at preventing adhesion of P. aeruginosa, Burkholderia cenocepacia, Burkholderia pseudomallei or L. pneumophila. This has been documented previously with dextran in P. aeruginosa (Bargouthi et al., 1996; Bryan et al., 1999) and Burkholderia cenocepacia (Chiu et al., 2001). Heparin has been shown to reduce P. aeruginosa colonization of contact lenses (Duran et al., 1993) and also to inhibit the adhesion of bacteria to epithelial cells (Plotkowski et al., 2001) and basement membrane collagen IV (Tsang et al., 2003). The polymeric saccharides are less effective at inhibiting adhesion of Bacillus anthracis and Y. pestis with the exception of dextran sulphate in Bacillus anthracis. It is likely that Bacillus anthracis attachment is inhibited more effectively by dextran sulphate than dextran because of the negative charge of this polysaccharide. That heparin is both negatively charged and a poor inhibitor may indicate that saccharide structure is also important for interaction with Bacillus anthracis adhesins.

The sulphated polymeric saccharides, heparin and dextran sulphate may have therapeutic benefits other than inhibition of bacterial attachment. Pathogenic yersiniae utilize a type III secretion system (Ysc) to deliver toxins and enzymes (Yops) into host cells. Exogenous heparin and dextran sulphate interact with LcrG from Y. enterocolitica and decrease the level of YopE translocation into host cells (Boyd et al., 1998). Similar interactions may occur in Y. pestis. Nebulized heparin has been shown to thin the mucus layer in Burkholderia-cenocepacia-colonized cystic fibrosis patients due to electrostatic interactions. In addition, heparin had an anti-inflammatory effect, reducing sputum and sera levels of IL6 and IL8 (Ledson et al., 2001). It has been reported that bacterial attachment can be mediated by hydrophobic interactions that can be interfered with by hydrophobic aromatic compounds such as p-nitrophenol (Falkowski et al., 1986). In this study, hydrophobic interactions were important for attachment because all of the bacteria, with the exception of Bacillus anthracis, were strongly inhibited by p-nitrophenol.

The sialylated saccharides, 3-sialyllactose and 6-sialyllactose, were good inhibitors of Burkholderia cenocepacia, L. pneumophila and P. aeruginosa attachment (Table 1). In contrast, mannosylated saccharides, with the exception of mannose and Man{alpha}1-2Man, were less effective inhibitors. The fucosylated saccharides, 2-fucosyllactose and 3-fucosyllactose only inhibited Burkholderia cenocepacia attachment and this was dependent on the Galß1-4Glc region rather than just the terminal Fuc{alpha}1-2Gal moiety. The effectiveness of digalactosyl saccharides (Gal{alpha}1-3Gal, Gal{alpha}1-4Gal and Galß1-4Gal) was linkage-dependent. Burkholderia cenocepacia and L. pneumophila preferred the {alpha}-linkage over the ß-linkage. In contrast, P. aeruginosa attachment was not inhibited by any digalactosyl saccharides. It has been demonstrated previously that the Gal{alpha}1-4Gal-containing glycolipids, digalactosylceramide and globotriosylceramide, can bind Burkholderia cenocepacia (Sylvester et al., 1996). Other common trends were observed. Disaccharides containing Gal linked to either GalNAc or GlcNAc by a ß1-3 or ß1-4 linkage were more inhibitory than if present with the corresponding ß1-6 linkage. This was particularly the case for P. aeruginosa. The observation that GalNAcß1-4Gal was amongst the best anti-adhesion compounds for each bacterium is interesting. This disaccharide is a component of the gangliosides, asialo-GM1 and asialo-GM2, demonstrated previously to bind a range of pathogens that cause respiratory disease (Table 3). For most of these bacteria GalNAcß1-4Gal has been determined as the minimum structure required for binding. Therefore, with the exception of Mycoplasma pneumoniae (Krivan et al., 1989), Mycobacterium tuberculosis, Coxiella burnetti and Chlamydia psittaci, all of the major causes of bacterial respiratory infection in humans have been demonstrated to bind the GalNAcß1-4Gal moiety in asialo-GM1. This raises the possibility of the creation of anti-adhesion compounds that can inhibit a range of bacterial pathogens and warrants further investigation.

Oligosaccharide specificity in relation to bacterial adhesins

The particular adhesins involved in the interactions with these saccharides are largely unknown. This highlights a significant gap in the understanding of the interaction of these bacteria with host cells. However, the literature contains details of various adhesins that may be involved in interactions with the respiratory epithelium. Burkholderia cenocepacia expresses cable pili (Cbl) that allows attachment to cytokeratin 13 (Sajjan et al., 2000). However a Cbl mutant was demonstrated to bind to A549 cells at a similar level to the wild-type strain (Tomich & Mohr, 2003), indicating that other mechanisms mediate attachment to this cell line. Pili have been observed that react with antibodies raised against the pili protein (PilA) from P. aeruginosa (Kuehn et al., 1992). Such pili may allow attachment to cell surface oligosaccharides. Indeed, the P. aeruginosa PilA protein has been demonstrated to bind GalNAcß1-4Gal (Schweizer et al., 1998), and may have a similar tropism in Burkholderia cenocepacia. Interestingly, Burkholderia pseudomallei adhesion was inhibited by ~90 % by a broad range of saccharide structures (Table 2). Galactose, glucose and N-acetylgalactosamine (1 mg ml–1) have been demonstrated previously to inhibit attachment, along with asialo-GM1 (12.5 µg ml–1) confirming the binding action of this glycolipid (Gori et al., 1999). In contrast to this study, mannose was not shown to inhibit attachment, however the concentration used was 25-fold less. It is well established that asialo-GM1 and asialo-GM2 are host cell receptors for Burkholderia pseudomallei (Kanai et al., 1997; Gori et al., 1999), however, the adhesins they bind have not been characterized. Possible adhesins identified include the capsule, LPS and flagellum (Atkins & Oyston, 2003). The completion of the genome sequence will lead to the identification of further adhesin candidates such as pili and non-pilus adhesins (www.sanger.ac.uk/Projects/B_pseudomallei). The flagellum has been demonstrated to mediate adhesion to amoebae and subsequent invasion (Inglis et al., 2003). The structure the flagellum attaches to, and whether a similar strategy is used to attach and invade human cells, are unknown.

Little is known about adhesion of vegetative Bacillus anthracis cells to human tissues. Putative adhesins include the S-layer proteins (EA1 and Sap), teichoic acids, the poly-{gamma}-D-glutamic acid capsule and fibronectin- and collagen-binding proteins (Ariel et al., 2003; Ezzell & Welkos, 1999; Fouet et al., 1999). The carbohydrate specificities displayed in this study (Table 2) were not due to interactions with the capsule, because the strain used lacked the pXO2 plasmid that encodes this virulence factor. Rather the S-layer and other putative adhesins, such as teichoic acid, are the likely candidates for such interactions. It cannot be discounted that expression of the capsule would alter the inhibition profile of this bacterium. Similarly, it should be recognized that in the event of pneumonic disease, such as would occur in the case of bioterrorism, initial interactions with the host cell would involve the B. anthracis spore prior to germination (Cieslak & Eitzen, 2000; Leggiadro, 2000). The spore may bind to entirely different structures from the vegetative cell and requires further investigation. The exosporium is a hexagonal lattice with filamentous appendages and glycoproteins (Fox et al., 2003; Mock & Fouet, 2001; Sylvestre et al., 2002) that may provide a mechanism for spore attachment to host cells prior to macrophage internalization.

This investigation details the potential of oligosaccharides to provide generic therapeutic benefits by inhibiting the attachment of a range of bacterial pathogens based on their pathological location, in this case the respiratory tract. Oligosaccharide inhibition of Burkholderia pseudomallei attachment to alveolar epithelial cells is particularly striking. Some of these compounds will be the subject of an in vivo investigation to determine the potential therapeutic benefit during intranasal and aerosol infections in a murine melioidosis model. As a generic therapy, the theory could be extended to include viruses. Sialylated glycoconjugates and sulphated dextrans have been shown to inhibit the attachment of various respiratory viruses to cell lines (Hosoya et al., 1991; Neyts et al., 1995; Suzuki et al., 1992; Suzuki et al., 2001). Theoretically, mixtures composed of saccharides such as 3-sialyllactose, polymeric saccharides and GalNAcß1-4Gal could afford protection against a spectrum of bacterial and viral respiratory pathogens, and may offer a novel prophylaxis.


   ACKNOWLEDGEMENTS
TOP
 INTRODUCTION
 METHODS
 RESULTS AND DISCUSSION
 ACKNOWLEDGEMENTS
REFERENCES
 
R. T. and T. B. recognize the contribution of Ministry of Defence funding for this work. Thank you to Professor Rick Titball and Dr Andrew Simpson for critical review of the manuscript.


   REFERENCES
TOP
 INTRODUCTION
 METHODS
 RESULTS AND DISCUSSION
 ACKNOWLEDGEMENTS
 REFERENCES
 

  • Ahmed, K., Suzuki, Y., Miyamoto, D. & Nagatake, T. (2002). Asialo-GM1 and asialo-GM2 are putative adhesion molecules for Moraxella catarrhalis. Med Microbiol Immunol 191, 5–10.[CrossRef][Medline]

  • Ariel, N., Zvi, A., Makarova, K. S., Chitlaru, T., Elhanany, E., Velan, B., Cohen, S., Friedlander, A. M. & Shafferman, A. (2003). Genome-based bioinformatic selection of chromosomal Bacillus anthracis putative vaccine candidates coupled with proteomic identification of surface-associated antigens. Infect Immun 71, 4563–4579.[Abstract/Free Full Text]

  • Atkins, T. P. & Oyston, P. C. F. (2003). The molecular basis of pathogenicity of Burkholderia pseudomallei. In Recent Research Developments in Infection & Immunity, vol. 1, pp. 1–8. Kerala, India: Transworld Research Network.

  • Bargouthi, S., Guerdoud, L. M. & Speert, D. P. (1996). Inhibition by dextran of Pseudomonas aeruginosa adherence to epithelial cells. Am J Respir Crit Care Med 154, 1788–1793.[Abstract]

  • Boyd, A. P., Sory, M.-P., Iriarte, M. & Cornelis, G. R. (1998). Heparin interferes with translocation of Yop proteins into HeLa cells and binds to LcrG, a regulatory component of the Yersinia Yop apparatus. Mol Microbiol 27, 425–436.[CrossRef][Medline]

  • Brennan, M. J., Hannah, J. H. & Leininger, E. (1991). Adhesion of Bordetella pertussis to sulfatides and to the GalNAcß1-4Gal sequence found in glycosphingolipids. J Biol Chem 266, 18827–18831.[Abstract/Free Full Text]

  • Bryan, R., Feldman, M., Jawetz, S. C., Rajan, S., DiMango, E., Tang, H. B., Scheffler, L., Speert, D. P. & Prince, A. (1999). The effects of aerosolized dextran in a mouse model of Pseudomonas aeruginosa pulmonary infection. J Infect Dis 179, 1449–1458.[CrossRef][Medline]

  • Chiu, C. H., Wong, S., Hancock, R. E. & Speert, D. P. (2001). Adherence of Burkholderia cepacia to respiratory tract epithelial cells and inhibition with dextrans. Microbiology 147, 2651–2658.[Abstract/Free Full Text]

  • Cieslak, T. J. & Eitzen, E. M. Jr (2000). Bioterrorism: agents of concern. J Public Health Manag Pract 6, 19–29.

  • Cundell, D. R. & Tuomanen, E. I. (1994). Receptor specificity of adherence of Streptococcus pneumoniae to human type-II pneumocytes and vascular endothelial cells in vitro. Microb Pathog 17, 361–374.[CrossRef][Medline]

  • Duran, J. A., Malvar, A., Rodriguez-Ares, M. T. & Garcia-Riestra, C. (1993). Heparin inhibits Pseudomonas adherence to soft contact lens. Eye 7, 152–154.

  • Edén, C. S., Freter, R., Hagberg, L., Hull, R., Hull, S., Leffler, H. & Schoolnik, G. (1982). Inhibition of experimental ascending urinary tract infection by an epithelial cell-surface receptor analogue. Nature 298, 560–563.[CrossRef][Medline]

  • Ezzell, J. W. & Welkos, S. L. (1999). The capsule of Bacillus anthracis, a review. J Appl Microbiol 87, 250. 250.[CrossRef][Medline]

  • Falkowski, W., Edwards, M. & Schaeffer, A. J. (1986). Inhibitory effect of substituted aromatic hydrocarbons on adherence of Escherichia coli to human epithelial cells. Infect Immun 52, 863–866.[Abstract/Free Full Text]

  • Fouet, A., Mesnage, S., Tosi-Couture, E., Gounon, P. & Mock, M. (1999). Bacillus anthracis surface: capsule and S-layer. J Appl Microbiol 87, 251–255.[CrossRef][Medline]

  • Fox, A., Stewart, G. C., Waller, L. N., Fox, K. F., Harley, W. M. & Price, R. L. (2003). Carbohydrates and glycoproteins of Bacillus anthracis and related bacilli: targets for biodetection. J Microbiol Methods 54, 143–152.[CrossRef][Medline]

  • Gori, A. H., Ahmed, K., Martinez, G., Masaki, H., Watanabe, K. & Nagatake, T. (1999). Mediation of attachment of Burkholderia pseudomallei to human pharyngeal epithelial cells by the asialoganglioside GM1-GM2 receptor complex. Am J Trop Med Hyg 61, 473–475.[Abstract]

  • Hosoya, M., Balzarini, J., Shigeta, S. & De Clercq, E. (1991). Differential inhibitory effects of sulfated polysaccharides and polymers on the replication of various myxoviruses and retroviruses, depending on the composition of the target amino acid sequences of the viral envelope glycoproteins. Antimicrob Agents Chemother 35, 2515-2520.[Abstract/Free Full Text]

  • Idänpään-Heikkilä, I., Simon, P. M., Zopf, D., Vullo, T., Cahill, P., Sokol, K. & Tuomanen, E. (1997). Oligosaccharides interfere with the establishment and progression of experimental pneumococcal pneumonia. J Infect Dis 176, 704–712.[Medline]

  • Inglis, T. J. J., Robertson, T., Woods, D. E., Dutton, N. & Chang, B. J. (2003). Flagellum-mediated adhesion by Burkholderia pseudomallei precedes invasion of Acanthamoeba astronyxis. Infect Immun 71, 2280–2282.[Abstract/Free Full Text]

  • Kanai, K., Suzuki, Y., Kondo, E., Maejima, Y., Miyamoto, D., Suzuki, T. & Kurata, T. (1997). Specific binding of Burkholderia pseudomallei cells and their cell-surface acid phosphatase to gangliotetraosylceramide (asialo GM1) and gangliotriaosylceramide (asialo GM2). Southeast Asian J Trop Med Public Health 28, 781–790.[Medline]

  • Karlsson, K.-A., Ångström, J., Bergström, J. & Lanne, B. (1992). Microbial interaction with animal cell surface carbohydrates. APMIS Suppl 27, 71–83.[Medline]

  • Kienle, Z., Emody, L., Svanborg, C. & O'Toole, P. W. (1992). Adhesive properties conferred by the plasminogen activator to Yersinia pestis. J Gen Microbiol 138, 1679–1687.

  • Krivan, H. C., Roberts, D. D. & Ginsburg, V. (1988a). Many pulmonary pathogenic bacteria bind specifically to the carbohydrate sequence GalNAcß1-4Gal found in some glycolipids. Proc Natl Acad Sci U S A 85, 6157–6161.[Abstract/Free Full Text]

  • Krivan, H. C., Ginsburg, V. & Roberts, D. D. (1988b). Pseudomonas aeruginosa and Pseudomonas cepacia isolated from cystic fibrosis patients bind specifically to gangliotetraosylceramide (asialo GM1) and gangliotriaosylceramide (asialo GM2). Arch Biochem Biophys 260, 493–496.[CrossRef][Medline]

  • Krivan, H. C., Olson, L. D., Barile, M. F., Ginsburg, V. & Roberts, D. D. (1989). Adhesion of Mycoplasma pneumoniae to sulfated glycolipids and inhibition by dextran sulfate. J Biol Chem 264, 9283–9288.[Abstract/Free Full Text]

  • Krivan, H. C., Nilsson, B., Lingwood, C. A. & Ryu, H. (1991). Chlamydia trachomatis and Chlamydia pneumoniae bind specifically to phosphatidylethanolamine in HeLa cells and to GalNAcß1-4Galß1-4Glc sequences found in asialo-GM1 and asialo-GM2. Biochem Biophys Res Comm 175, 1082–1089.[CrossRef][Medline]

  • Kuehn, M., Lent, K., Haas, J., Hagenzieker, J., Cervin, M. & Smith, A. L. (1992). Fimbriation of Pseudomonas cepacia. Infect Immun 60, 2002–2007.[Abstract/Free Full Text]

  • Kwaik, Y. A. (2000). Invasion of mammalian and protozoan cells by Legionella pneumophila. Subcell Biochem 33, 383–409.[Medline]

  • Lähteenmäki, K., Virkola, R., Sarén, A., Emödy, L. & Korhonen, T. K. (1998). Expression of plasminogen activator Pla of Yersinia pestis enhances bacterial attachment to the mammalian extracellular matrix. Infect Immun 66, 5755–5762.[Abstract/Free Full Text]

  • Ledson, M., Gallagher, M., Hart, C. A. & Walshaw, M. (2001). Nebulized heparin in Burkholderia cepacia colonized adult cystic fibrosis patients. Eur Respir J 17, 36–38.[Abstract/Free Full Text]

  • Leggiadro, R. J. (2000). The threat of biological terrorism: a public health and infection control reality. Infect Control Hosp Epidemiol 21, 53–56.[CrossRef][Medline]

  • Lindler, L. E. & Tall, B. D. (1993). Yersinia pestis pH 6 antigen forms fimbriae and is induced by intracellular association with macrophages. Mol Microbiol 8, 311–324.[Medline]

  • Lister, P. D. (2000). Emerging resistance problems among respiratory tract pathogens. Am J Manag Care 6 (Suppl 8), S409–S418.[Medline]

  • Loomes, L. M., Uemura, K., Childs, R. A. & 7 other authors (1984). Erythrocyte receptors for Mycoplasma pneumoniae are sialylated oligosaccharides of Ii antigen types. Nature 307, 560–563.[CrossRef][Medline]

  • Lynch, J. P. III (2001). Hospital-acquired pneumonia: risk factors, microbiology, and their treatment. Chest 119 (Suppl 2), 373S–384S.[Abstract/Free Full Text]

  • Mock, M. & Fouet, A. (2001). Anthrax. Annu Rev Microbiol 55, 647–671.[CrossRef][Medline]

  • Neyts, J., Reyman, D., Letourneur, D. & 9 other authors (1995). Differential antiviral activity of derivatized dextrans. Biochem Pharmacol 50, 743–751.[CrossRef][Medline]

  • Ofek, I. & Sharon, N. (1990). Adhesins as lectins: specificity and role in infection. Curr Top Microbiol Immunol 151, 91–113.[Medline]

  • Payne, D., Tatham, D., Williamson, E. D. & Titball, R. W. (1998). The pH 6 antigen of Yersinia pestis binds to ß1-linked galactosyl residues in glycosphingolipids. Infect Immun 66, 4545–4548.[Abstract/Free Full Text]

  • Plotkowski, M. C., Costa, A. O., Morandi, V., Barbosa, H. S., Nader, H. B., de Bentzmann, S. & Puchelle, E. (2001). Role of heparan sulphate proteoglycans as potential receptors for non-piliated Pseudomonas aeruginosa adherence to non-polarised airway epithelial cells. J Med Microbiol 50, 183–190.[Abstract/Free Full Text]

  • Ramphal, R., Carnoy, C., Fievre, S., Michalski, J.-C., Houdret, N., Lamblin, G., Strecker, G. & Roussel, P. (1991). Pseudomonas aeruginosa recognizes carbohydrate chains containing type 1 (Galß1-3GlcNAc) or type 2 (Galß1-4GlcNAc) disaccharide units. Infect Immun 59, 700–704.[Abstract/Free Full Text]

  • Rojas, C. M., Ham, J. H., Deng, W. L., Doyle, J. J. & Collmer, A. (2002). HecA, a member of a class of adhesins produced by diverse pathogenic bacteria, contributes to the attachment, aggregation, epidermal cell killing, and virulence phenotypes of Erwinia chrysanthemi EC16 on Nicotiana clevelandii seedlings. Proc Natl Acad Sci U S A 99, 13142–13147.[Abstract/Free Full Text]

  • Sajjan, U. S., Sylvester, F. A. & Forstner, J. F. (2000). Cable-piliated Burkholderia cepacia binds to cytokeratin 13 of epithelial cells. Infect Immun 68, 1787–1795.[Abstract/Free Full Text]

  • Schweizer, F., Jiao, H., Hindsgaul, O., Wong, W. Y. & Irvin, R. T. (1997). Interaction between the pili of Pseudomonas aeruginosa PAK and its carbohydrate receptor ß-D-GalNAc(1->4)ß-D-Gal analogs. Can J Microbiol 44, 307–311.

  • Simon, P. M., Goode, P. L., Mobasseri, A. & Zopf, D. (1997). Inhibition of Helicobacter pylori binding to gastrointestinal epithelial cells by sialic acid-containing oligosaccharides. Infect Immun 65, 750–757.[Abstract]

  • Spencer, R. C. (1995). The emergence of epidemic, multiple antibiotic resistant Stenotrophomonas (Xanthomonas) maltophila and Burkholderia (Pseudomonas) cepacia. J Hosp Infect 30, S453–S464.[CrossRef]

  • Straley, S. C. (1993). Adhesins in Yersinia pestis. Trends Microbiol 1, 285–286.[CrossRef][Medline]

  • Sundberg-Kovamees, M., Holme, T. & Sjogren, A. (1994). Specific binding of Streptococcus pneumoniae to two receptor saccharide structures. Microb Pathog 17, 63–68.[CrossRef][Medline]

  • Suzuki, Y., Nakao, T., Ito, T. & 14 other authors (1992). Structural determination of gangliosides that bind to influenza A, B, and C viruses by an improved binding assay: strain-specific receptor epitopes in sialo-sugar chains. Virology 189, 121–131.[CrossRef][Medline]

  • Suzuki, T., Portner, A., Scroggs, R. A., Uchikawa, M., Koyama, N., Matsuo, K., Suzuki, Y. & Takimoto, T. (2001). Receptor specificities of human respiroviruses. J Virol 75, 4604–4613.[Abstract/Free Full Text]

  • Sylvester, F. A., Sajjan, U. S. & Forstner, J. F. (1996). Burkholderia (basonym Pseudomonas) cepacia binding to lipid receptors. Infect Immun 64, 1420–1425.[Abstract]

  • Sylvestre, P., Couture-Tosi, E. & Mock, M. (2002). A collagen-like surface glycoprotein is a structural component of the Bacillus anthracis exosporium. Mol Microbiol 45, 169–178.[CrossRef][Medline]

  • Tan, T. Q. (2003). Antibiotic resistant infections due to Streptococcus pneumoniae: impact on therapeutic options and clinical outcome. Curr Opin Infect Dis 16, 271–277.[Medline]

  • Thomas, R. J. & Brooks, T. J. (2004). Oligosaccharide receptor mimics inhibit Legionella pneumophila attachment to human respiratory epithelial cells. Microb Pathog 36, 83–92.[CrossRef][Medline]

  • Tomich, M. & Mohr, C. D. (2003). Adherence and autoaggregation phenotype of a Burkholderia cenocepacia cable pilus mutant. FEMS Microbiol Lett 228, 287–297.[CrossRef][Medline]

  • Tong, H. H., McIver, M. A., Fisher, L. M. & DeMaria, T. F. (1999). Effect of lacto-N-neotetraose, asialoganglioside-GM1 and neuraminidase on adherence of otitis media-associated serotypes of Streptococcus pneumoniae to chinchilla tracheal epithelium. Microb Pathog 26, 111–119.[CrossRef][Medline]

  • Tsang, K. W., Shum, D. K., Chan, S. & 7 other authors (2003). Pseudomonas aeruginosa adherence to human basement membrane collagen in vitro. Eur Respir J 21, 932–938.[Abstract/Free Full Text]

  • van Alphen, L., Geelen-van den Broek, L., Blaas, L., van Ham, M. & Dankert, J. (1991). Blocking of fimbria-mediated adherence of Haemophilus influenzae by sialyl gangliosides. Infect Immun 59, 4473–4477.[Abstract/Free Full Text]

  • Zopf, D. & Roth, S. (1996). Oligosaccharide anti-infective agents. Lancet 347, 1017–1021.[CrossRef][Medline]




    This article has been cited by other articles:


    		Home page
    		 MicrobiologyHome page
    M. Drasbek, G. Christiansen, K. R. Drasbek, A. Holm, and S. Birkelund
    Interaction between the P1 protein of Mycoplasma pneumoniae and receptors on HEp-2 cells
    Microbiology, November 1, 2007; 153(11): 3791 - 3799.
    [Abstract] [Full Text] [PDF]

    		Home page
    		 J Med MicrobiolHome page
    A. P. Anisimov and K. K. Amoako
    Treatment of plague: promising alternatives to antibiotics.
    J. Med. Microbiol., November 1, 2006; 55(Pt 11): 1461 - 1475.
    [Abstract] [Full Text] [PDF]

    		Home page
    		 Infect. Immun.Home page
    E. Behling-Kelly, H. Vonderheid, K. S. Kim, L. B. Corbeil, and C. J. Czuprynski
    Roles of Cellular Activation and Sulfated Glycans in Haemophilus somnus Adherence to Bovine Brain Microvascular Endothelial Cells
    Infect. Immun., September 1, 2006; 74(9): 5311 - 5318.
    [Abstract] [Full Text] [PDF]

    		Home page
    		 Infect. Immun.Home page
    J. A. Boddey, C. P. Flegg, C. J. Day, I. R. Beacham, and I. R. Peak
    Temperature-Regulated Microcolony Formation by Burkholderia pseudomallei Requires pilA and Enhances Association with Cultured Human Cells
    Infect. Immun., September 1, 2006; 74(9): 5374 - 5381.
    [Abstract] [Full Text] [PDF]

    		Home page
    		 J Med MicrobiolHome page
    R. Thomas and T. Brooks
    Attachment of Yersinia pestis to human respiratory cell lines is inhibited by certain oligosaccharides.
    J. Med. Microbiol., March 1, 2006; 55(Pt 3): 309 - 315.
    [Abstract] [Full Text] [PDF]

    This Article
    Right arrow Abstract Freely available
    Right arrow Full Text (PDF)
    Right arrow Alert me when this article is cited
    Right arrow Alert me if a correction is posted
    Right arrow Citation Map
    Services
    Right arrow Email this article to a friend
    Right arrow Similar articles in this journal
    Right arrow Similar articles in PubMed
    Right arrow Alert me to new issues of the journal
    Right arrow Download to citation manager
    Right arrow reprints & permissions
    Citing Articles
    Right arrow Citing Articles via HighWire
    Right arrow Citing Articles via CrossRef
    Right arrow Citing Articles via Google Scholar
    Google Scholar
    Right arrow Articles by Thomas, R.
    Right arrow Articles by Brooks, T.
    Right arrow Search for Related Content
    PubMed
    Right arrow PubMed Citation
    Right arrow Articles by Thomas, R.
    Right arrow Articles by Brooks, T.
    Agricola
    Right arrow Articles by Thomas, R.
    Right arrow Articles by Brooks, T.

    HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
    INT J SYST EVOL MICROBIOL J MED MICROBIOL MICROBIOLOGY J GEN VIROL ALL SGM JOURNALS