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. 1986 Feb;51(2):461–469. doi: 10.1128/iai.51.2.461-469.1986

Multiplication of Shigella flexneri within HeLa cells: lysis of the phagocytic vacuole and plasmid-mediated contact hemolysis.

P J Sansonetti, A Ryter, P Clerc, A T Maurelli, J Mounier
PMCID: PMC262354  PMID: 3510976

Abstract

Virulent strains of Shigella flexneri invade HeLa cells with high efficiency. This crucial step in the pathogenic process is encoded by a 140-megadalton plasmid which induces phagocytosis of the bacteria by host cells. In this report we used pWR100, the virulence plasmid of S. flexneri serotype 5, and pHS4108, a 32-megadalton subclone of pWR100, to demonstrate that the plasmid is also responsible for rapid intracellular growth of the bacteria. The ability to replicate intracellularly was not correlated with induction of Shiga toxin. However, plasmid-mediated intracellular multiplication was strongly correlated with the ability of the bacteria to rapidly and efficiently lyse the phagocytic vacuole and replicate freely in the cytoplasm. Temperature-regulated plasmid-mediated contact hemolytic activity strongly correlated with both phagosomal membrane lysis and efficient intracellular multiplication. We propose this virulence plasmid-associated hemolysin to be an important factor in the invasion and proliferation of Shigella spp. in mammalian cells.

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Selected References

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  1. Brown J. E., Rothman S. W., Doctor B. P. Inhibition of protein synthesis in intact HeLa cells by Shigella dysenteriae 1 toxin. Infect Immun. 1980 Jul;29(1):98–107. doi: 10.1128/iai.29.1.98-107.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. FORMAL S. B., KUNDEL D., SCHNEIDER H., KUNEVN, SPRINZ H. Studies with Vibrio cholerae in the ligated loop of the rabbit intestine. Br J Exp Pathol. 1961 Oct;42:504–510. [PMC free article] [PubMed] [Google Scholar]
  3. FURNESS G., ROWLEY D. Transduction of virulence within the species Salmonella typhimurium. J Gen Microbiol. 1956 Aug;15(1):140–145. doi: 10.1099/00221287-15-1-140. [DOI] [PubMed] [Google Scholar]
  4. Formal S. B., Hale T. L., Sansonetti P. J. Invasive enteric pathogens. Rev Infect Dis. 1983 Sep-Oct;5 (Suppl 4):S702–S707. doi: 10.1093/clinids/5.supplement_4.s702. [DOI] [PubMed] [Google Scholar]
  5. Gentry M. K., Dalrymple J. M. Quantitative microtiter cytotoxicity assay for Shigella toxin. J Clin Microbiol. 1980 Sep;12(3):361–366. doi: 10.1128/jcm.12.3.361-366.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hale T. L., Bonventre P. F. Shigella infection of Henle intestinal epithelial cells: role of the bacterium. Infect Immun. 1979 Jun;24(3):879–886. doi: 10.1128/iai.24.3.879-886.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hale T. L., Formal S. B. Protein synthesis in HeLa or Henle 407 cells infected with Shigella dysenteriae 1, Shigella flexneri 2a, or Salmonella typhimurium W118. Infect Immun. 1981 Apr;32(1):137–144. doi: 10.1128/iai.32.1.137-144.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hale T. L., Morris R. E., Bonventre P. F. Shigella infection of henle intestinal epithelial cells: role of the host cell. Infect Immun. 1979 Jun;24(3):887–894. doi: 10.1128/iai.24.3.887-894.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Labrec E. H., Schneider H., Magnani T. J., Formal S. B. EPITHELIAL CELL PENETRATION AS AN ESSENTIAL STEP IN THE PATHOGENESIS OF BACILLARY DYSENTERY. J Bacteriol. 1964 Nov;88(5):1503–1518. doi: 10.1128/jb.88.5.1503-1518.1964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Le Minor S., Le Coueffic E. Etude sur les hémolysines des enterobacteriaceae. Ann Microbiol (Paris) 1975 Oct-Nov;126(3):313–332. [PubMed] [Google Scholar]
  11. Maurelli A. T., Baudry B., d'Hauteville H., Hale T. L., Sansonetti P. J. Cloning of plasmid DNA sequences involved in invasion of HeLa cells by Shigella flexneri. Infect Immun. 1985 Jul;49(1):164–171. doi: 10.1128/iai.49.1.164-171.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Maurelli A. T., Blackmon B., Curtiss R., 3rd Temperature-dependent expression of virulence genes in Shigella species. Infect Immun. 1984 Jan;43(1):195–201. doi: 10.1128/iai.43.1.195-201.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Moulder J. W. Intracellular parasitism: life in an extreme environment. J Infect Dis. 1974 Sep;130(3):300–306. doi: 10.1093/infdis/130.3.300. [DOI] [PubMed] [Google Scholar]
  14. O'Brien A. D., LaVeck G. D., Thompson M. R., Formal S. B. Production of Shigella dysenteriae type 1-like cytotoxin by Escherichia coli. J Infect Dis. 1982 Dec;146(6):763–769. doi: 10.1093/infdis/146.6.763. [DOI] [PubMed] [Google Scholar]
  15. Rikihisa Y., Ito S. Intracellular localization of Rickettsia tsutsugamushi in polymorphonuclear leukocytes. J Exp Med. 1979 Sep 19;150(3):703–708. doi: 10.1084/jem.150.3.703. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. SERENY B. Experimental keratoconjunctivitis shigellosa. Acta Microbiol Acad Sci Hung. 1957;4(4):367–376. [PubMed] [Google Scholar]
  17. Sansonetti P. J., Hale T. L., Dammin G. J., Kapfer C., Collins H. H., Jr, Formal S. B. Alterations in the pathogenicity of Escherichia coli K-12 after transfer of plasmid and chromosomal genes from Shigella flexneri. Infect Immun. 1983 Mar;39(3):1392–1402. doi: 10.1128/iai.39.3.1392-1402.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Sansonetti P. J., Kopecko D. J., Formal S. B. Involvement of a plasmid in the invasive ability of Shigella flexneri. Infect Immun. 1982 Mar;35(3):852–860. doi: 10.1128/iai.35.3.852-860.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Sansonetti P. J., Kopecko D. J., Formal S. B. Shigella sonnei plasmids: evidence that a large plasmid is necessary for virulence. Infect Immun. 1981 Oct;34(1):75–83. doi: 10.1128/iai.34.1.75-83.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Sansonetti P. J., d'Hauteville H., Ecobichon C., Pourcel C. Molecular comparison of virulence plasmids in Shigella and enteroinvasive Escherichia coli. Ann Microbiol (Paris) 1983 May-Jun;134A(3):295–318. [PubMed] [Google Scholar]
  21. Takeuchi A. Electron microscope studies of experimental Salmonella infection. I. Penetration into the intestinal epithelium by Salmonella typhimurium. Am J Pathol. 1967 Jan;50(1):109–136. [PMC free article] [PubMed] [Google Scholar]
  22. Takeuchi A., Sprinz H., LaBrec E. H., Formal S. B. Experimental bacillary dysentery. An electron microscopic study of the response of the intestinal mucosa to bacterial invasion. Am J Pathol. 1965 Dec;47(6):1011–1044. [PMC free article] [PubMed] [Google Scholar]
  23. Winkler H. H., Miller E. T. Phospholipase A activity in the hemolysis of sheep and human erythrocytes by Rickettsia prowazeki. Infect Immun. 1980 Aug;29(2):316–321. doi: 10.1128/iai.29.2.316-321.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Winkler H. H., Miller E. T. Phospholipase A and the interaction of Rickettsia prowazekii and mouse fibroblasts (L-929 cells). Infect Immun. 1982 Oct;38(1):109–113. doi: 10.1128/iai.38.1.109-113.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]

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