1887

Microbiology Society logo

Volume 147, Issue 3

exerts a cytolethal distending toxin effect on HeLa cells and T lymphocytes Free

Abstract

is an emerging human enteropathogen. However, little is known about the pathogenesis of infection. In this study the authors demonstrate that whole-cell preparations and extracts produce a cytolethal distending toxin (CDT)-like effect on HeLa cells characterized by progressive distension and nuclear fragmentation culminating in cell death over 5 d. To further delineate the nature of this toxic effect in relation to CDT from other pathogens, the effect of on cellular events in epithelial cells and immunocytes was investigated. lysate-treated HeLa cells subjected to FACScan analysis using carboxyfluorescein diacetete succinimidyl ester (CFDA-SE) as a cell tracer demonstrated cell division arrest. Propidium iodide (PI) staining of HeLa cells revealed that cell cycle arrest occurred in G/M. Human T lymphocytes exposed to lysates also showed cell cycle arrest in G/M. Using a combination of Annexin V/PI staining and TUNEL assay, cytodistended HeLa cells were shown to undergo apoptotic cell death. These data provide the first insights into the virulence mechanisms of this novel enteropathogen.

  • Received:
  • Accepted:
  • Revised:
  • Published Online:
Keyword(s): apoptosis , carboxyfluorescein succinimidyl ester , CDT , CDT, cytolethal distending toxin , CFDA-SE, carboxyfluorescein diacetete succinimidyl ester , CFSE , enteropathogen , PHA, phytohaemagglutinin , PI, propidium iodide and virulence factors
© Microbiology Society
Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-147-3-735
2001-03-01
2024-04-24
Loading full text...

Full text loading...

/deliver/fulltext/micro/147/3/1470735a.html?itemId=/content/journal/micro/10.1099/00221287-147-3-735&mimeType=html&fmt=ahah

References

  1. Albert M. J., Faruque S. M., Faruque A. S., Bettelheim K. A., Neogi P. K., Bhuiyan N. A., Kaper J. B. 1996; Controlled study of cytolethal distending toxin-producing Escherichia coli infections in Bangladeshi children. J Clin Microbiol 34:717–719
     [Google Scholar]
  2. Aragon V., Chao K., Dreyfus L. A. 1997; Effect of cytolethal distending toxin on F-actin assembly and cell division in Chinese hamster ovary cells. Infect Immun 65:3774–3780
     [Google Scholar]
  3. Bourke B., Sherman P., Louie H., Hani E., Islur P., Chan V. L. 1995; Physical and genetic map of the genome of Campylobacter upsaliensis . Microbiology 141:2417–2424 [CrossRef]
     [Google Scholar]
  4. Bourke B., Bingham H. L., Chan V. L, al Rashid S. T. 1996; Characterization of Campylobacter upsaliensis fur and its localization in a highly conserved region of the Campylobacter genome. Gene 183:219–224 [CrossRef]
     [Google Scholar]
  5. Bourke B., Chan V. L., Sherman P. 1998; Campylobacter upsaliensis : waiting in the wings. Clin Microbiol Rev 11:440–449
     [Google Scholar]
  6. Carter J. E., Cimolai N. 1996; Hemolytic-uremic syndrome associated with acute Campylobacter upsaliensis gastroenteritis. Nephron 74:489 [CrossRef]
     [Google Scholar]
  7. Chusid M. J., Wortmann D. W., Dunne W. M. 1990; ‘ Campylobacter upsaliensis ’ sepsis in a boy with acquired hypogammaglobulinemia. Diagn Microbiol Infect Dis 13:367–369 [CrossRef]
     [Google Scholar]
  8. Comayras C., Tasca C., Peres S. Y., Ducommun B., Oswald E., De Rycke J. 1997; Escherichia coli cytolethal distending toxin blocks the HeLa cell cycle at the G2/M transition by preventing cdc2 protein kinase dephosphorylation and activation. Infect Immun 65:5088–5095
     [Google Scholar]
  9. Cope L. D., Lumbley S., Latimer J. L. 8 other authors 1997; A diffusible cytotoxin of Haemophilus ducreyi. Proc Natl Acad Sci USA 94:4056–4061 [CrossRef]
     [Google Scholar]
  10. De Rycke J., Mazars P., Nougayrede J. P., Tasca C., Boury M., Herault F., Valette A., Oswald E. 1996; Mitotic block and delayed lethality in HeLa epithelial cells exposed to Escherichia coli BM2-1 producing cytotoxic necrotizing factor type 1. Infect Immun 64:1694–1705
     [Google Scholar]
  11. DeRycke J., Sert V., Comayras C., Tasca C. 2000; Sequence of lethal events in HeLa cells exposed to the G2 blocking cytolethal distending toxin. Eur J Cell Biol 79:192–201 [CrossRef]
     [Google Scholar]
  12. Gelfanova V., Hansen E. J., Spinola S. M. 1999; Cytolethal distending toxin of Haemophilus ducreyi induces apoptotic death of Jurkat T cells. Infect Immun 67:6394–6402
     [Google Scholar]
  13. Goddard E. A., Lastovica A. J., Argent A. C. 1997; Campylobacter O: 41 isolation in Guillain–Barre syndrome. Arch Dis Child 76:526–528 [CrossRef]
     [Google Scholar]
  14. Hickey T. E., McVeigh S. A., Carroll S. A., Bourgeois A. L., Guerry P. 1999; Biological properties of cytolethal distending toxin. 10th International Workshop on Campylobacter, Helicobacter and Related Organisms Baltimore, MD: abstract CP34;
     [Google Scholar]
  15. Ho T. W., Hsieh S. T., Nachamkin I. 8 other authors 1997; Motor nerve terminal degeneration provides a potential mechanism for rapid recovery in acute motor axonal neuropathy after Campylobacter infection. Neurology 48:717–724 [CrossRef]
     [Google Scholar]
  16. Jenkin G. A., Tee W. 1998; Campylobacter upsaliensis -associated diarrhea in human immunodeficiency virus-infected patients. Clin Infect Dis 27:816–821 [CrossRef]
     [Google Scholar]
  17. Jimenez S. G., Heine R. G., Ward P. B., Robins-Browne R. M. 1999; Campylobacter upsaliensis gastroenteritis in childhood. Pediatr Infect Dis J 18:988–992 [CrossRef]
     [Google Scholar]
  18. Johnson W. M., Lior H. 1988; A new heat-labile cytolethal distending toxin (CLDT) produced by Campylobacter spp. Microb Pathog 4:115–126 [CrossRef]
     [Google Scholar]
  19. Kato S., Muro M., Akifusa S., Hanada N., Semba I., Fujii T., Kowashi Y., Nishihara T. 1995; Evidence for apoptosis of murine macrophages by Actinobacillus actinomycetemcomitans infection. Infect Immun 63:3914–3919
     [Google Scholar]
  20. Lastovica A. J., LeRoux E. 1993; Prevalence of Campylobacter spp. in the diarrhoeic stools and blood cultures of pediatric patients. Acta Gastroenterol Belg 56:suppl34
     [Google Scholar]
  21. Lindblom G. B., Sjogren E., Hansson-Westerberg J., Kaijser B. 1995; Campylobacter upsaliensis , C. sputorum sputorum and C. concisus as common causes of diarrhoea in Swedish children. Scand J Infect Dis 27:187–188 [CrossRef]
     [Google Scholar]
  22. Musmanno R. A., Russi M., Figura N., Guglielmetti P., Zanchi A., Signori R., Rossolini A. 1998; Unusual species of campylobacters isolated in the Siena Tuscany area, Italy. New Microbiol 21:15–22
     [Google Scholar]
  23. Okuda J., Kurazono H., Takeda Y. 1995; Distribution of the cytolethal distending toxin A gene ( cdtA ) among species of Shigella and Vibrio , and cloning and sequencing of the cdt gene from Shigella dysenteriae . Microb Pathog 18:167–172 [CrossRef]
     [Google Scholar]
  24. Okuda J., Fukumoto M., Takeda Y., Nishibuchi M. 1997; Examination of diarrheagenicity of cytolethal distending toxin: suckling mouse response to the products of the cdtABC genes of Shigella dysenteriae . Infect Immun 65:428–433
     [Google Scholar]
  25. Patton C. M., Shaffer N., Edmonds P., Barrett T. J., Lambert M. A., Baker C., Perlman D. M., Brenner D. J. 1989; Human disease associated with ‘ Campylobacter upsaliensis ’ (catalase-negative or weakly positive Campylobacter species) in the United States. J Clin Microbiol 27:66–73
     [Google Scholar]
  26. Peres S. Y., Marches O., Daigle F., Nougayrede J. P., Herault F., Tasca C., De Rycke J., Oswald E. 1997; A new cytolethal distending toxin (CDT) from Escherichia coli producing CNF2 blocks HeLa cell division in G2/M phase. Mol Microbiol 24:1095–1107 [CrossRef]
     [Google Scholar]
  27. Pickett C. L., Pesci E. C., Cottle D. L., Russell G., Erdem A. N., Zeytin H. 1996; Prevalence of cytolethal distending toxin production in Campylobacter jejuni and relatedness of Campylobacter sp. cdtB gene. Infect Immun 64:2070–2078
     [Google Scholar]
  28. Purdy D., Buswell C. M., Hodgson A. E., McAlpine K., Henderson I., Leach S. A. 2000; Characterisation of cytolethal distending toxin (CDT) mutants of Camplylobacter jejuni . J Med Microbiol 49:473–479
     [Google Scholar]
  29. Scott D. A., Kaper J. B. 1994; Cloning and sequencing of the genes encoding Escherichia coli cytolethal distending toxin. Infect Immun 62:244–251
     [Google Scholar]
  30. Shenker B. J., McKay T., Datar S. 7 other authors 1999; Actinobacillus actinomycetemcomitans immunosuppressive protein is a member of the family of cytolethal distending toxins capable of causing a G2 arrest in human T cells. J Immunol 162:4773–4780
     [Google Scholar]
  31. Steele T. W., Sangster N., Lanser J. A. 1985; DNA relatedness and biochemical features of Campylobacter spp. isolated in central and South Australia. J Clin Microbiol 22:71–74
     [Google Scholar]
  32. Sugai M., Kawamoto T., Peres S. Y., Ueno Y., Komatsuzawa H., Fujiwara T., Kurihara H., Suginaka H., Oswald E. 1998; The cell cycle-specific growth-inhibitory factor produced by Actinobacillus actinomycetemcomitans is a cytolethal distending toxin. Infect Immun 66:5008–5019
     [Google Scholar]
  33. Sylvester F. A., Philpott D., Gold B., Lastovica A., Forstner J. F. 1996; Adherence to lipids and intestinal mucin by a recently recognized human pathogen, Campylobacter upsaliensis . Infect Immun 64:4060–4066
     [Google Scholar]
  34. Whitehouse C. A., Balbo P. B., Pesci E. C., Cottle D. L., Mirabito P. M., Pickett C. L. 1998; Campylobacter jejuni cytolethal distending toxin causes a G2-phase cell cycle block. Infect Immun 66:1934–1940
     [Google Scholar]
  35. Zhu J., Meinersmann R. J., Hiett K. L., Evans D. L. 1999; Apoptotic effect of outer-membrane proteins from Campylobacter jejuni on chicken lymphocytes. Curr Microbiol 38:244–249 [CrossRef]
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-147-3-735
Loading
Campylobacter upsaliensis exerts a cytolethal distending toxin effect on HeLa cells and T lymphocytes
Microbiology 147, 735 (2001); https://doi.org/10.1099/00221287-147-3-735
/content/journal/micro/10.1099/00221287-147-3-735
/content/journal/micro/10.1099/00221287-147-3-735
Loading

Data & Media loading...

Most cited Most Cited RSS feed

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