Abstract
Two quantitative, rapid assays were developed to study the adhesion of Actinobacillus actinomycetemcomitans, an oral bacterium associated with periodontal disease, to human epithelial cells. The human oral carcinoma cell line KB was grown in microtiter plates, and adherent bacteria were detected by an enzyme-linked immunosorbent assay with purified anti-A. actinomycetemcomitans serum and horseradish peroxidase-conjugated secondary antibody or [3H]thymidine-labeled bacteria. Adhesion was found to be time dependent and increased linearly with increasing numbers of bacteria added. Variation in the level of adhesion was noted among strains of A. actinomycetemcomitans. Adhesion was not significantly altered by changes in pH (from pH 5 to 9) but was sensitive to sodium chloride concentrations greater than 0.15 M. Pooled human saliva was inhibitory for adhesion when bacteria were pretreated with saliva before being added to the cells. Pretreatment of the KB cells with saliva did not inhibit adhesion. Protease treatment of A. actinomycetemcomitans reduced adhesion of the bacteria to KB cells. The data are consistent with the hypothesis that a protein(s) is required for bacterial adhesion and that host components may play a role in modulating adhesion to epithelial cells.
Full text
PDFImages in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Beachey E. H. Bacterial adherence: adhesin-receptor interactions mediating the attachment of bacteria to mucosal surface. J Infect Dis. 1981 Mar;143(3):325–345. doi: 10.1093/infdis/143.3.325. [DOI] [PubMed] [Google Scholar]
- Duncan M. J., Nakao S., Skobe Z., Xie H. Interactions of Porphyromonas gingivalis with epithelial cells. Infect Immun. 1993 May;61(5):2260–2265. doi: 10.1128/iai.61.5.2260-2265.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Elder B. L., Boraker D. K., Fives-Taylor P. M. Whole-bacterial cell enzyme-linked immunosorbent assay for Streptococcus sanguis fimbrial antigens. J Clin Microbiol. 1982 Jul;16(1):141–144. doi: 10.1128/jcm.16.1.141-144.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Finlay B. B., Falkow S. Common themes in microbial pathogenicity. Microbiol Rev. 1989 Jun;53(2):210–230. doi: 10.1128/mr.53.2.210-230.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fives-Taylor P. M., Thompson D. W. Surface properties of Streptococcus sanguis FW213 mutants nonadherent to saliva-coated hydroxyapatite. Infect Immun. 1985 Mar;47(3):752–759. doi: 10.1128/iai.47.3.752-759.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gibbons R. J. Bacterial adhesion to oral tissues: a model for infectious diseases. J Dent Res. 1989 May;68(5):750–760. doi: 10.1177/00220345890680050101. [DOI] [PubMed] [Google Scholar]
- Kagermeier A. S., London J. Actinobacillus actinomycetemcomitans strains Y4 and N27 adhere to hydroxyapatite by distinctive mechanisms. Infect Immun. 1985 Mar;47(3):654–658. doi: 10.1128/iai.47.3.654-658.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kiley P., Holt S. C. Characterization of the lipopolysaccharide from Actinobacillus actinomycetemcomitans Y4 and N27. Infect Immun. 1980 Dec;30(3):862–873. doi: 10.1128/iai.30.3.862-873.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kusters J. G., Mulders-Kremers G. A., van Doornik C. E., van der Zeijst B. A. Effects of multiplicity of infection, bacterial protein synthesis, and growth phase on adhesion to and invasion of human cell lines by Salmonella typhimurium. Infect Immun. 1993 Dec;61(12):5013–5020. doi: 10.1128/iai.61.12.5013-5020.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
- Meyer D. H., Fives-Taylor P. M. Characteristics of adherence of Actinobacillus actinomycetemcomitans to epithelial cells. Infect Immun. 1994 Mar;62(3):928–935. doi: 10.1128/iai.62.3.928-935.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Meyer D. H., Fives-Taylor P. M. Evidence that extracellular components function in adherence of Actinobacillus actinomycetemcomitans to epithelial cells. Infect Immun. 1993 Nov;61(11):4933–4936. doi: 10.1128/iai.61.11.4933-4936.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Meyer D. H., Sreenivasan P. K., Fives-Taylor P. M. Evidence for invasion of a human oral cell line by Actinobacillus actinomycetemcomitans. Infect Immun. 1991 Aug;59(8):2719–2726. doi: 10.1128/iai.59.8.2719-2726.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mintz K. P., Mann K. G. Detection of procollagen biosynthesis using peptide-specific antibodies. Matrix. 1990 Jul;10(3):186–199. doi: 10.1016/s0934-8832(11)80168-x. [DOI] [PubMed] [Google Scholar]
- Moore W. E. Microbiology of periodontal disease. J Periodontal Res. 1987 Sep;22(5):335–341. doi: 10.1111/j.1600-0765.1987.tb01595.x. [DOI] [PubMed] [Google Scholar]
- Nowotny A., Behling U. H., Hammond B., Lai C. H., Listgarten M., Pham P. H., Sanavi F. Release of toxic microvesicles by Actinobacillus actinomycetemcomitans. Infect Immun. 1982 Jul;37(1):151–154. doi: 10.1128/iai.37.1.151-154.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ofek I., Courtney H. S., Schifferli D. M., Beachey E. H. Enzyme-linked immunosorbent assay for adherence of bacteria to animal cells. J Clin Microbiol. 1986 Oct;24(4):512–516. doi: 10.1128/jcm.24.4.512-516.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Preus H. R., Namork E., Olsen I. Fimbriation of Actinobacillus actinomycetemcomitans. Oral Microbiol Immunol. 1988 Jun;3(2):93–94. doi: 10.1111/j.1399-302x.1988.tb00090.x. [DOI] [PubMed] [Google Scholar]
- Robertson P. B., Lantz M., Marucha P. T., Kornman K. S., Trummel C. L., Holt S. C. Collagenolytic activity associated with Bacteroides species and Actinobacillus actinomycetemcomitans. J Periodontal Res. 1982 May;17(3):275–283. doi: 10.1111/j.1600-0765.1982.tb01154.x. [DOI] [PubMed] [Google Scholar]
- Rosan B., Slots J., Lamont R. J., Listgarten M. A., Nelson G. M. Actinobacillus actinomycetemcomitans fimbriae. Oral Microbiol Immunol. 1988 Jun;3(2):58–63. doi: 10.1111/j.1399-302x.1988.tb00082.x. [DOI] [PubMed] [Google Scholar]
- Shenker B. J., Kushner M. E., Tsai C. C. Inhibition of fibroblast proliferation by Actinobacillus actinomycetemcomitans. Infect Immun. 1982 Dec;38(3):986–992. doi: 10.1128/iai.38.3.986-992.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Slots J., Genco R. J. Black-pigmented Bacteroides species, Capnocytophaga species, and Actinobacillus actinomycetemcomitans in human periodontal disease: virulence factors in colonization, survival, and tissue destruction. J Dent Res. 1984 Mar;63(3):412–421. doi: 10.1177/00220345840630031101. [DOI] [PubMed] [Google Scholar]
- Slots J., Listgarten M. A. Bacteroides gingivalis, Bacteroides intermedius and Actinobacillus actinomycetemcomitans in human periodontal diseases. J Clin Periodontol. 1988 Feb;15(2):85–93. doi: 10.1111/j.1600-051x.1988.tb00999.x. [DOI] [PubMed] [Google Scholar]
- Slots J., Reynolds H. S., Genco R. J. Actinobacillus actinomycetemcomitans in human periodontal disease: a cross-sectional microbiological investigation. Infect Immun. 1980 Sep;29(3):1013–1020. doi: 10.1128/iai.29.3.1013-1020.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Slots J. Salient Biochemical Characters of Actinobacillus actinomycetemcomitans. Arch Microbiol. 1982 Feb;131(1):60–67. doi: 10.1007/BF00451500. [DOI] [PubMed] [Google Scholar]
- Socransky S. S. Microbiology of periodontal disease -- present status and future considerations. J Periodontol. 1977 Sep;48(9):497–504. doi: 10.1902/jop.1977.48.9.497. [DOI] [PubMed] [Google Scholar]
- Sreenivasan P. K., Meyer D. H., Fives-Taylor P. M. Factors influencing the growth and viability of Actinobacillus actinomycetemcomitans. Oral Microbiol Immunol. 1993 Dec;8(6):361–369. doi: 10.1111/j.1399-302x.1993.tb00612.x. [DOI] [PubMed] [Google Scholar]
- Sreenivasan P. K., Meyer D. H., Fives-Taylor P. M. Requirements for invasion of epithelial cells by Actinobacillus actinomycetemcomitans. Infect Immun. 1993 Apr;61(4):1239–1245. doi: 10.1128/iai.61.4.1239-1245.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stanislawski L., Simpson W. A., Hasty D., Sharon N., Beachey E. H., Ofek I. Role of fibronectin in attachment of Streptococcus pyogenes and Escherichia coli to human cell lines and isolated oral epithelial cells. Infect Immun. 1985 Apr;48(1):257–259. doi: 10.1128/iai.48.1.257-259.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sweet S. P., MacFarlane T. W., Samaranayake L. P. An in vitro method to study the adherence of oral bacteria to HeLa cells. Microbios. 1989;60(242):15–22. [PubMed] [Google Scholar]
- Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tsai C. C., Shenker B. J., DiRienzo J. M., Malamud D., Taichman N. S. Extraction and isolation of a leukotoxin from Actinobacillus actinomycetemcomitans with polymyxin B. Infect Immun. 1984 Feb;43(2):700–705. doi: 10.1128/iai.43.2.700-705.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Van Dyke T. E., Bartholomew E., Genco R. J., Slots J., Levine M. J. Inhibition of neutrophil chemotaxis by soluble bacterial products. J Periodontol. 1982 Aug;53(8):502–508. doi: 10.1902/jop.1982.53.8.502. [DOI] [PubMed] [Google Scholar]
- Winkler J. R., John S. R., Kramer R. H., Hoover C. I., Murray P. A. Attachment of oral bacteria to a basement-membrane-like matrix and to purified matrix proteins. Infect Immun. 1987 Nov;55(11):2721–2726. doi: 10.1128/iai.55.11.2721-2726.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Yeung M. K., Cisar J. O. Sequence homology between the subunits of two immunologically and functionally distinct types of fimbriae of Actinomyces spp. J Bacteriol. 1990 May;172(5):2462–2468. doi: 10.1128/jb.172.5.2462-2468.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zambon J. J. Actinobacillus actinomycetemcomitans in human periodontal disease. J Clin Periodontol. 1985 Jan;12(1):1–20. doi: 10.1111/j.1600-051x.1985.tb01348.x. [DOI] [PubMed] [Google Scholar]