1887

Abstract

The fate of GAS and epithelial cells following internalization was determined in this study. HEp-2 cells harbouring intracellular bacteria were treated with antibiotics to kill extracellular adherent bacteria, washed, and the fate of bacteria and epithelial cells was assessed up to 24 h post-infection. In the absence of antibiotics, massive bacterial growth was apparent in the cell medium, accompanied by extensive cell death, suggesting that intracellular bacteria had multiplied and damaged the monolayer. Addition of the internalization inhibitor, cytochalasin D, either pre- or post-internalization prevented bacterial growth and cell injury; post-internalization treatment with chloramphenicol had the same effect. Analysis of three apoptotic markers in HEp-2 cells – chromatin condensation, DNA laddering and translocation of phosphatidylserine onto the cell-surface membrane – indicated that HEp-2 cells underwent apoptosis. Taken together, the data presented here support a model in which intenalized bacteria can induce their own externalization into the medium by a process that requires both an intact host-cell cytoskeleton and synthesis of bacterial proteins. Concomitantly, intracellular and, apparently, extracellular free bacteria induce apoptosis through their cytotoxic activity, and release essential nutrients required for their growth.

Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.05263-0
2004-01-01
2024-04-24
Loading full text...

Full text loading...

/deliver/fulltext/jmm/53/1/JM530101.html?itemId=/content/journal/jmm/10.1099/jmm.0.05263-0&mimeType=html&fmt=ahah

References

  1. Bayles K. W., Wesson C. A., Liou L. E., Fox L. K., Bohach G. A., Trumble W. R. 1998; Intracellular Staphylococcus aureus escapes the endosome and induces apoptosis in epithelial cells. Infect Immun 66:336–342
    [Google Scholar]
  2. Bricker A. L., Cywes C., Ashbaugh C. D., Wessels M. R. 2002; NAD+-glycohydrolase acts as an intracellular toxin to enhance the extracellular survival of group A streptococci. Mol Microbiol 44:257–269 [CrossRef]
    [Google Scholar]
  3. Cunningham M. W. 2000; Pathogenesis of group A streptococcal infections. Clin Microbiol Rev 13:470–511 [CrossRef]
    [Google Scholar]
  4. Grassme H., Jendrossek V., Gulbins E. 2001; Molecular mechanisms of bacteria induced apoptosis. Apoptosis 6:441–445 [CrossRef]
    [Google Scholar]
  5. Greco R., De Martino L., Donnarumma G., Conte M. P., Seganti L., Valenti P. 1995; Invasion of cultured human cells by Streptococcus pyogenes . Res Microbiol 146:551–560 [CrossRef]
    [Google Scholar]
  6. Jadoun J., Ozeri V., Burstein E., Skutelsky E., Hanski E., Sela S. 1998; Protein F1 is required for efficient entry of Streptococcus pyogenes into epithelial cells. J Infect Dis 178:147–158 [CrossRef]
    [Google Scholar]
  7. Kahl B. G., Goulian M., van Wamel W., Herrmann M., Simon S. M., Kaplan G., Peters G., Cheung A. L. 2000; Staphylococcus aureus RN6390 replicates and induces apoptosis in a pulmonary epithelial cells. Infect Immun 68:5385–5392 [CrossRef]
    [Google Scholar]
  8. Kim J. M., Eckmann L., Savidge T. C., Lowe D. C., Witthoft T., Kagnoff M. F. 1998; Apoptosis of human intestinal epithelial cells after bacterial invasion. J Clin Invest 102:1815–1823 [CrossRef]
    [Google Scholar]
  9. Kuo C., Wu J., Tsai P., Kao F., Lei H., Lin M., Lin Y. 1999; Streptococcal pyrogenic exotoxin B induces apoptosis and reduces phagocytic activity in U937 cells. Infect Immun 67:126–130
    [Google Scholar]
  10. LaPenta D., Rubens C., Chi E., Cleary P. P. 1994; Group A streptococci efficiently invade human respiratory epithelial cells. Proc Natl Acad Sci U S A 91:12115–12119 [CrossRef]
    [Google Scholar]
  11. Marouni M. J., Sela S. 2004; Intracellular survival of persistent group A streptococci in cultured epithelial cells. Int J Med Microbiol (in press
    [Google Scholar]
  12. Menzies B. E., Kourteva I. 1998; Internalization of Staphylococcus aureus by endothelial cells induces apoptosis. Infect Immun 66:5994–5998
    [Google Scholar]
  13. Molinari G., Chhatwal G. S. 1998; Invasion and survival of Streptococcus pyogenes in eukaryotic cells correlates with the source of the clinical isolates. J Infect Dis 177:1600–1607 [CrossRef]
    [Google Scholar]
  14. Molinari G., Chhatwal G. S. 1999; Streptococcal invasion. Curr Opin Microbiol 2:56–61 [CrossRef]
    [Google Scholar]
  15. Molinari G., Rohde M., Guzman C. A., Chhatwal G. S. 2000; Two distinct pathways for the invasion of Streptococcus pyogenes in non-phagocytic cells. Cell Microbiol 2:145–154 [CrossRef]
    [Google Scholar]
  16. Molinari G., Rohde M., Talay S. R., Chhatwal G. S., Beckert S., Podbielski A. 2001; The role played by the group A streptococcal negative regulator Nra on bacterial interactions with epithelial cells. Mol Microbiol 40:99–114 [CrossRef]
    [Google Scholar]
  17. Nakagawa I., Nakata M., Kawabata S., Hamada S. 2001; Cytochrome c -mediated caspase-9 activation triggers apoptosis in Streptococcus pyogenes -infected epithelial cells. Cell Microbiol 3:395–405 [CrossRef]
    [Google Scholar]
  18. Neeman R., Keller N., Barzilai A., Korenman Z., Sela S. 1998; Prevalence of internalization-associated gene, prtF1 , among persisting group-A streptococcus strains isolated from asymptomatic carriers. Lancet 352:1974–1977 [CrossRef]
    [Google Scholar]
  19. Nuzzo I., Sanges M. R., Folgore A., Carratelli C. R. 2000; Apoptosis of human keratinocytes after bacterial invasion. FEMS Immunol Med Microbiol 27:235–240 [CrossRef]
    [Google Scholar]
  20. Österlund A., Engstrand L. 1995; Intracellular penetration and survival of Streptococcus pyogenes in respiratory epithelial cells in vitro. Acta Otolaryngol 115:685–688 [CrossRef]
    [Google Scholar]
  21. Österlund A., Engstrand L. 1997; An intracellular sanctuary for Streptococcus pyogenes in human tonsillar epithelium – studies of asymptomatic carriers and in vitro cultured biopsies. Acta Otolaryngol 117:883–888 [CrossRef]
    [Google Scholar]
  22. Österlund A., Popa R., Nikkila T., Scheynius A., Engstrand L. 1997; Intracellular reservoir of Streptococcus pyogenes in vivo: a possible explanation for recurrent pharyngotonsillitis. Laryngoscope 107:640–647 [CrossRef]
    [Google Scholar]
  23. Raffray M., Cohen G. M. 1997; Apoptosis and necrosis in toxicology : a continuum or distinct modes of cell death?. Pharmacol Ther 75:153–177 [CrossRef]
    [Google Scholar]
  24. Schrager H. M., Rheinwald J. G., Wessels M. R. 1996; Hyaluronic acid capsule and the role of streptococcal entry into keratinocytes in invasive skin infection. J Clin Invest 98:1954–1958 [CrossRef]
    [Google Scholar]
  25. Sela S., Barzilai A. 1999; Why do we fail with penicillin in the treatment of group A streptococcus infections?. Ann Med 31:303–307 [CrossRef]
    [Google Scholar]
  26. Sela S., Neeman R., Keller N., Barzilai A. 2000; Relationship between asymptomatic carriage of Streptococcus pyogenes and the ability of the strains to adhere to and be internalised by cultured epithelial cells. J Med Microbiol 49:499–502
    [Google Scholar]
  27. Tsai P., Lin Y., Kuo C., Lei H., Wu J. 1999; Group A streptococcus induces apoptosis in human epithelial cells. Infect Immun 667:4334–4339
    [Google Scholar]
  28. Weinrauch Y., Zychlinsky A. 1999; The induction of apoptosis by bacterial pathogens. Annu Rev Microbiol 53:155–187 [CrossRef]
    [Google Scholar]
  29. Zychlinsky A., Sansonetti P. 1997; Apoptosis in bacterial pathogenesis. J Clin Invest 100:493–495 [CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/jmm.0.05263-0
Loading
/content/journal/jmm/10.1099/jmm.0.05263-0
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error