Log in

Intracellular degradation of Fusobacterium nucleatum in human gingival epithelial cells

  • Published:
Molecules and Cells

Abstract

The role of Fusobacterium nucleatum in oral health and disease is controversial. We have previously shown that F. nucleatum invades gingival epithelial cells. However, the destiny of the internalized F. nucleatum is not clear. In the present study, the intracellular destiny of F. nucleatum and its cytopathic effect on gingival epithelial cells were studied. The ability of F. nucleatum and seven other oral bacterial species to invade immortalized human gingival epithelial (HOK-16B) cells were compared by confocal microscopy and flow cytometry. F. nucleatum had the highest invasive capacity, comparable to that of Porphyromonas gingivalis, a periodontal pathogen. Confocal microscopic examination revealed colocalization of internalized F. nucleatum with endosomes and lysosomes. Examination by transmission electron microscopy revealed that most intracellular F. nucleatum was located within vesicular structures with single enclosed membranes. Furthermore, F. nucleatum could not survive within gingival epithelial cells and had no cytopathic effects on host cells. Interestingly, endosomal maturation played a role in induction of the antimicrobial peptides human beta defensin (HBD)-2 and -3 by F. nucleatum from gingival epithelial cells. F. nucleatum is destined to enter an endocytic degradation pathway after invasion and has no cytopathic effect on gingival epithelial cells, which may cast new light on the role of F. nucleatum in the pathogenesis of periodontitis.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Allenspach-Petrzilka, G.E., and Guggenheim, B. (1983). Bacterial invasion of the periodontium; an important factor in the pathogenesis of periodontitis? J. Clin. Periodontol. 10, 609–617.

    Article  CAS  PubMed  Google Scholar 

  • Alonso, A., and García-del Portillo, F. (2004). Hijacking of eukaryotic functions by intracellular bacterial pathogens. Int. Microbiol. 7, 181–191.

    PubMed  Google Scholar 

  • Amano, A., Nakagawa, L., and Yoshimori, T. (2006). Autophagy in innate immunity against intracellular bacteria. J. Biochem. 140, 161–166.

    Article  CAS  PubMed  Google Scholar 

  • Bachrach, G., Rosen, G., Bellalou, M., Naor, R., and Sela, M.N. (2004). Identification of a Fusobacterium nucleatum 65 kDa serine protease. Oral Microbiol. Immunol. 19, 155–159.

    Article  CAS  PubMed  Google Scholar 

  • Bauer, C., Schoonbroodt, D., Wagner, C., and Horsmans, Y. (2000). Liver abscesses due to Fusobacterium species. Liver 20, 267–268.

    Article  CAS  PubMed  Google Scholar 

  • Chaim, W., and Mazor, M. (1992). Intraamniotic infection with fusobacteria. Arch. Gynecol. Obstet. 251, 1–7.

    Article  CAS  PubMed  Google Scholar 

  • Chung, W.O., and Dale, B.A. (2004). Innate immune response of oral and foreskin keratinocytes: utilization of different signaling pathways by various bacterial species. Infect. Immun. 72, 352–358.

    Article  CAS  PubMed  Google Scholar 

  • Feng, Z., and Weinberg, A. (2006). Role of bacteria in health and disease of periodontal tissues. Periodontol. 2000 40, 50–76.

    Article  Google Scholar 

  • Fuller, J.R., Craven, R.R., Hall, J.D., Kijek, T.M., Taft-Benz, S., and Kawula, T.H. (2008). RipA, a cytoplasmic membrane protein conserved among Francisella species, is required for intracellular survival. Infec. Immun. 76, 4934–4943.

    Article  CAS  Google Scholar 

  • Grabenstein, J.P., Marceau, M., Pujol, C., Simonet, M., and Bliska, J.B. (2004). The response regulator PhoP of Yersinia pseudotu berculosis is important for replication in macrophages and for virulence. Infect. Immun. 72, 4973–4984.

    Article  CAS  PubMed  Google Scholar 

  • Gursoy, U.K., Könönen, E., and Uitto, V.J. (2008). Intracellular replication of fusobacteria requires new actin filament formation of epithelial cells. APMIS 116, 1063–1070.

    Article  PubMed  Google Scholar 

  • Han, Y.W., Shi, W., Huang, G.T., Kinder Haake, S., Park, N.H., Kuramitsu, H., and Genco, R.J. (2000). Interactions between periodontal bacteria and human oral epithelial cells: Fusobacterium nucleatum adheres to and invades epithelial cells. Infect. Immun. 68, 3140–3146.

    Article  CAS  PubMed  Google Scholar 

  • Han, Y.W., Redline, R.W., Li, M., Yin, L., Hill, G.B., and McCormick, T.S. (2004). Fusobacterium nucleatum induces premature and term stillbirths in pregnant mice: implication of oral bacteria in preterm birth. Infect. Immun. 72, 2272–2279.

    Article  CAS  PubMed  Google Scholar 

  • Hanada, H., Moriyama, Y., Maeda, M., and Futai, M. (1999). Kinetic studies of chromaffin granule H+-ATPase and effects of bafilomycin A1. Biochem. Biophys. Res. Commun. 170, 873–878.

    Article  Google Scholar 

  • Hintermann, E., Haake, S.K., Christen, U., Sharabi, A., and Quaranta, V. (2002). Discrete proteolysis of focal contact and adherens junction components in Porphyromonas gingivalis-infected oral keratinocytes: a strategy for cell adhesion and migration disabling. Infect. Immun. 70, 5846–5856.

    Article  CAS  PubMed  Google Scholar 

  • Hsu, C.Y., and Luh, K.T. (1995). Cytology of pulmonary Fusobacterium nucleatum infection. A case report. Acta Cytol. 39, 114–117.

    CAS  PubMed  Google Scholar 

  • Hultén, K., Rigo, R., Gustafsson, I., and Engstrand, L. (1996). New pharmacokinetic in vitro model for studies of antibiotic activity against intracellular microorganisms. Antimicrob. Agents Chemother. 40, 2727–2731.

    PubMed  Google Scholar 

  • Ji, S., Hyun, J., Park, E., Lee, B.L., Kim, K.K., and Choi, Y. (2007a). Susceptibility of various oral bacteria to antimicrobial peptides and to phagocytosis by neutrophils. J. Periodont. Res. 42, 410–419.

    Article  CAS  PubMed  Google Scholar 

  • Ji, S., Kim, Y., Min, B-Y., Han, S.H., and Choi, Y. (2007b). Innate immune responses of gingival epithelial cells to nonperiodontopathic and periodontopathic bacteria. J. Periodont. Res. 42, 503–510.

    Article  CAS  PubMed  Google Scholar 

  • Ji, S., Shin, J.E., Kim, Y.S., Oh, J.E., Min, B.M., and Choi, Y. (2009). Toll-like receptor 2 and NALP2 mediate induction of human beta-defensins by Fusobacterium nucleatum in gingival epithelial cells. Infect. Immun. 77, 1044–1052.

    Article  CAS  PubMed  Google Scholar 

  • Kim, Y.C., Ko, Y., Hong, S.-D., Kim, K.Y., Lee, Y.H., Chae, C., and Choi, Y. (2010). Presence of Porphyromonas gingivalis and plasma cell dominance in gingival tissues with periodontitis. Oral Dis. 16, 375–381.

    Article  CAS  PubMed  Google Scholar 

  • Krisanaprakornkit, S., Kimball, J.R., Weinberg, A., Darveau, R.P., Bainbridge, B.W., and Dale, B.A. (2000). Inducible expression of human beta-defensin 2 by Fusobacterium nucleatum in oral epithelial cells: multiple signaling pathways and role of commensal bacteria in innate immunity and the epithelial barrier. Infect. Immun. 68, 2907–2915.

    Article  CAS  PubMed  Google Scholar 

  • Lamont, R.J., Chan, A., Belton, C.M., Izutsu, K.T., Vasel, D., and Weinberg, A. (1995). Porphyromonas gingivalis invasion of gingival epithelial cells. Infect. Immun. 63, 3878–3885.

    CAS  PubMed  Google Scholar 

  • Lux, R., Miller, J.N., Park, N.H., and Shi, W. (2001). Motility and chemotaxis in tissue penetration of oral epithelial cell layers by Treponema denticola. Infect. Immun. 69, 6276–6283.

    Article  CAS  PubMed  Google Scholar 

  • Meyer, D.H., Lippmann, J.E., and Fives-Taylor, P.M. (1996). Invasion of epithelial cells by Actinobacillus actinomycetemcomitans: a dynamic, multistep process. Infect. Immun. 64, 2988–2997.

    CAS  PubMed  Google Scholar 

  • Park, N.-H., Min, B.-M., Li, S.L., Huang, M.Z., Cherick, H.M., and Doniger, J. (1991). Immortalization of normal human oral keratinocytes with type 16 human papillomavirus. Carcinogenesis 12, 1627–1631.

    Article  CAS  PubMed  Google Scholar 

  • Pils, S., Schmitter, T., Neske, F., and Hauck, C.R. (2006). Quantification of bacterial invasion into adherent cells by flow cytometry. J. Microbiol. Methods 65, 301–310.

    Article  CAS  PubMed  Google Scholar 

  • Rossano, F., Rizzo, A., Sangnes, M.R., Cipollaro de L’Ero, G., and Tufano, M.A. (1993). Human monocytes and gingival fibroblasts release tumor necrosis factor-a, interleukin-1a and interleukin-6 in response to particulate and soluble fractions of Prevotella melaninogenica and Fusobacterium nucleatum. Int. J. Clin. Res. 23, 165–168.

    Article  CAS  Google Scholar 

  • Saito, A., Inagaki, S., and Ishihara, K. (2009). Differential ability of periodontopathic bacteria to modulate invasion of human gingival epithelial cells by Porphyromonas gingivalis. Microb. Pathog. 47, 329–333.

    Article  CAS  PubMed  Google Scholar 

  • Socransky, S.S., Haffajee, A.D., Cugini, M.A., Smith, C., and Kent, R.L. Jr. (1998). Microbial complexes in subgingival plaque. J. Clin. Periodontol. 25, 134–144.

    Article  CAS  PubMed  Google Scholar 

  • Tricker, E., and Cheng, G. (2008). With a little help from my friends: modulation of phagocytosis through TLR activation. Cell Res. 18, 711–712.

    Article  CAS  PubMed  Google Scholar 

  • Yilmaz, O., Verbeke, P., Lamont, R.J., and Ojcius, D.M. (2006). Intercellular spreading of Porphyromonas gingivalis infection in primary gingival epithelial cells. Infect. Immun. 74, 703–710.

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Youngnim Choi.

About this article

Cite this article

Ji, S., Shin, J.E., Kim, Y.C. et al. Intracellular degradation of Fusobacterium nucleatum in human gingival epithelial cells. Mol Cells 30, 519–526 (2010). https://doi.org/10.1007/s10059-010-0142-8

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10059-010-0142-8

Keywords

Navigation