1887

Abstract

The Gram-negative periodontopathogen () binds selectively to buccal epithelial cells (BECs) of human and Old World primates by means of the outer-membrane autotransporter protein Aae. We speculated that the exposed N-terminal portion of the passenger domain of Aae would mediate binding to BECs. By using a series of plasmids that express full-length or truncated Aae proteins in , we found that the BEC-binding domain of Aae was located in the N-terminal surface-exposed region of the protein, specifically in the region spanning amino acids 201–284 just upstream of the repeat region within the passenger domain. Peptides corresponding to amino acids 201–221, 222–238 and 201–240 were synthesized and tested for their ability to reduce Aae-mediated binding to BECs based on results obtained with truncated Aae proteins expressed in . BEC-binding of expressing Aae was reduced by as much as 50 % by pre-treatment of BECs with a 40-mer peptide (201–240; P40). Aae was also shown to mediate binding to cultured human epithelial keratinocytes (TW2.6), OBA9 and TERT, and endothelial (HUVEC) cells. Pre-treatment of epithelial cells with P40 resulted in a dose-dependent reduction in binding and reduced the binding of both full-length and truncated Aae proteins expressed in , as well as Aae expressed in . Fluorescently labelled P40 peptides reacted in a dose-dependent manner with BEC receptors. We propose that these proof-of-principle experiments demonstrate that peptides can be designed to interfere with binding mediated by host-cell receptors specific for Aae adhesins.

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2010-11-01
2020-04-10
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References

  1. Aronson M., Medalia O., Schori L., Mirelman D., Sharon N., Ofek I.. 1979; Prevention of colonization of the urinary tract of mice with Escherichia coli by blocking of bacterial adherence with methyl α-d-mannopyranoside. J Infect Dis139:329–332
    [Google Scholar]
  2. Asakawa R., Komatsuzawa H., Kuwai T., Yamada S., Goncalves R. B., Izumi S., Fujiwara T., Nakano Y., Suzuki N.. other authors 2003; Outer membrane protein 100, a versatile virulence factor of Actinobacillus actinomycetemcomitans. Mol Microbiol50:1125–1139
    [Google Scholar]
  3. Beachey E. H.. 1981; Bacterial adherence: adhesin–receptor interactions mediating the attachment of bacteria to mucosal surfaces. J Infect Dis143:325–345
    [Google Scholar]
  4. Beachey E. H., Courtney H. S.. 1987; Bacterial adherence: the attachment of group A streptococci to mucosal surfaces. Rev Infect Dis9:S475–S481
    [Google Scholar]
  5. Cheney C. P., Schad P. A., Formal S. B., Boedeker E. C.. 1980; Species specificity of in vitro Escherichia coli adherence to host intestinal cell membranes and its correlation with in vivo colonization and infectivity. Infect Immun28:1019–1027
    [Google Scholar]
  6. Eger T., Zoller L., Muller H.-P., Hoffmann S., Lobinsky D.. 1996; Potential diagnostic value of sampling oral mucosal surfaces for Actinobacillus acintomycetemcomitans in young adults. Eur J Oral Sci104:112–117
    [Google Scholar]
  7. Fine D. H., Furgang D., Schreiner H. C., Goncharoff P., Charlesworth J., Ghazwan G., Fitzgerald-Bocarsly P., Figurski D. H.. 1999; Phenotypic variation in Actinobacillus actinomycetemcomitans during laboratory growth: implications for virulence. Microbiology145:1335–1347
    [Google Scholar]
  8. Fine D. H., Velliyagounder K., Furgang D., Kaplan J. B.. 2005; The Actinobacillus actinomycetemcomitans autotransporter adhesin Aae exhibits specificity for buccal epithelial cells from humans and Old World primates. Infect Immun73:1947–1953
    [Google Scholar]
  9. Fine D. H., Kaplan J. B., Kachlany S. C., Schreiner H. C.. 2006; How we got attached to Actinobacillus actinomycetemcomitans: a model for infectious diseases. Periodontol 2000;42:114–157
    [Google Scholar]
  10. Fine D. H., Markowitz K., Furgang D., Fairlie K., Ferrandiz J., Nasri C., McKiernan M., Gunsolley J.. 2007; Aggregatibacter actinomycetemcomitans and its relationship to initiation of localized aggressive periodontitis: a longitudinal cohort study of initially healthy adolescents. J Clin Microbiol45:3859–3869
    [Google Scholar]
  11. Fives-Taylor P. M., Meyer D. H., Mintz K. P., Brissette C.. 1999; Virulence factors of Actinobacillus actinomycetemcomitans. Periodontol 2000;20:136–167
    [Google Scholar]
  12. Haraszthy V. I., Zambon J. J., Trevisan M., Zeid M., Genco R. J.. 2000; Identification of periodontal pathogens in atheromatous plaques. J Periodontol71:1554–1560
    [Google Scholar]
  13. Haubek D., Ennibi O.-K., Poulsen P., Vaeth M., Kilian K.. 2008; Risk of aggressive periodontitis in adolescent carriers of the JP2 clone of Aggregatibacter ( Actinobacillus) actinomycetemcomitans in Morocco: a prospective longitudinal cohort study. Lancet371:237–242
    [Google Scholar]
  14. Henderson I. R., Nataro J. P.. 2001; Virulence functions of autotransporter proteins. Infect Immun69:1231–1243
    [Google Scholar]
  15. Henderson I. R., Navarro-Garcia F., Nataro J. P.. 1998; The great escape: structure and function of the autotransporter proteins. Trends Microbiol6:370–378
    [Google Scholar]
  16. Henderson B., Wilson M., Sharp L., Ward J. M.. 2002; Actinobacillus actinomycetemcomitans. J Med Microbiol51:1013–1020
    [Google Scholar]
  17. Henderson I. R., Navarro-Garcia F., Desvaux M. J., Fernandez R. C., Ala'Aldeen D.. 2004; Type V protein secretion pathway: the autotransporter story. Microbiol Mol Biol Rev68:692–744
    [Google Scholar]
  18. Kok S. H., Hong C. Y., Lin S. K., Lee J. J., Chiang C. P., Kuo M. Y.. 2007; Establishment and characterization of a tumorigenic cell line from areca quid and tobacco smoke-associated buccal carcinoma. Oral Oncol43:639–647
    [Google Scholar]
  19. Komatsuzawa H., Asakawa R., Kawai T., Ochiai K., Fujiwara T., Taubman M. A., Ohara M., Kurihara H., Sugai M.. 2002; Identification of six major outer membrane proteins from Actinobacillus actinomycetemcomitans. Gene288:195–201
    [Google Scholar]
  20. Kozarov E. V., Dorn B. R., Shelburne C. E., Dunn W. A. Jr, Progulske-Fox A.. 2005; Human atherosclerotic plaque contains viable invasive Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis. Arterioscler Thromb Vasc Biol25:e17–e18
    [Google Scholar]
  21. Kusumoto Y., Hirano H., Saitoh K., Yamada S., Takedachi M., Nozaki T., Ozawa Y., Nakahira Y., Saho T.. other authors 2004; Human gingival epithelial cells produce chemotactic factors interleukin-8 and monocyte chemoattractant protein-1 after stimulation with Porphyromonas gingivalis via Toll-like receptor 2. J Periodontol75:370–379
    [Google Scholar]
  22. Müller H.-P., Zöller L., Eger T., Hoffmann S., Lobinsky D.. 1996; Natural distribution of oral Actinobacillus actinomycetemcomitans in young men with minimal periodontal disease. J Periodontal Res31:373–380
    [Google Scholar]
  23. Newman M. G., Socransky S. S., Savit E. D., Propas D. A., Crawford A.. 1976; Studies of the microbiology of periodontosis. J Periodontol47:373–379
    [Google Scholar]
  24. Rose J. E., Meyer D. H., Fives-Taylor P. M.. 2003; Aae, an autotransporter involved in adhesion of Actinobacillus actinomycetemcomitans to epithelial cells. Infect Immun71:2384–2393
    [Google Scholar]
  25. Rudney J. D., Chen R., Sedgewick G. J.. 2001; Intracellular Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis in buccal epithelial cells collected from human subjects. Infect Immun69:2700–2707
    [Google Scholar]
  26. Schenkein H. A., Barbour S. E., Berry C. R., Kipps B., Tew J. G.. 2000; Invasion of human vascular endothelial cells by Actinobacillus actinomycetemcomitans via the receptor for platelet-activating factor. Infect Immun68:5416–5419
    [Google Scholar]
  27. Sellwood R., Gibbons R. A., Jones G. W., Rutter J. M.. 1975; Adhesion of enteropathogenic Escherichia coli to pig intestinal brush borders: the existence of two pig phenotypes. J Med Microbiol8:405–411
    [Google Scholar]
  28. Simpson W. A., Beachey E. H.. 1983; Adherence of group A streptococci to fibronectin on oral epithelial cells. Infect Immun39:275–279
    [Google Scholar]
  29. Slots J.. 1976; The predominant cultivable organisms in juvenile periodontitis. Scand J Dent Res84:1–10
    [Google Scholar]
  30. Ukkonen P., Varis K., Jernfors M., Herva E., Jokinen J., Ruokokoski E., Zopf D., Kilpi T.. 2000; Treatment of acute otitis media with an antiadhesive oligosaccharide: a randomized, double-blind, placebo-controlled trial. Lancet356:1398–1402
    [Google Scholar]
  31. Yue G., Kaplan J. B., Furgang D., Mansfield K. G., Fine D. H.. 2007; A second Aggregatibacter actinomycetemcomitans autotransporter adhesin that exhibits specificity for buccal epithelial cells of humans and Old World primates. Infect Immun75:4440–4448
    [Google Scholar]
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