Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1989 Sep;57(9):2919–2922. doi: 10.1128/iai.57.9.2919-2922.1989

Internalization of Shigella flexneri into HeLa cells occurs without an increase in cytosolic Ca2+ concentration.

P L Clerc 1, B Berthon 1, M Claret 1, P J Sansonetti 1
PMCID: PMC313550  PMID: 2668195

Abstract

Pathogenic Shigella flexneri invades epithelial cells through directed phagocytosis. The role of intracellular free calcium ([Ca2+]i) as a signal for this process was tested. No variation in the [Ca2+]i could be detected by using the fluorescent indicator fura 2 to measure the [Ca2+]i in HeLa cells during the invasion process. In addition, neither clamping nor maximal decreasing or increasing of the [Ca2+]i of HeLa cells had any effect on their susceptibility to invasion. These data demonstrate that [Ca2+]i is not a signal for S. flexneri directed phagocytosis in HeLa cells.

Full text

PDF
2919

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Babior B. M. Oxygen-dependent microbial killing by phagocytes (first of two parts). N Engl J Med. 1978 Mar 23;298(12):659–668. doi: 10.1056/NEJM197803232981205. [DOI] [PubMed] [Google Scholar]
  2. Bainton D. F. Sequential degranulation of the two types of polymorphonuclear leukocyte granules during phagocytosis of microorganisms. J Cell Biol. 1973 Aug;58(2):249–264. doi: 10.1083/jcb.58.2.249. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Clerc P., Sansonetti P. J. Entry of Shigella flexneri into HeLa cells: evidence for directed phagocytosis involving actin polymerization and myosin accumulation. Infect Immun. 1987 Nov;55(11):2681–2688. doi: 10.1128/iai.55.11.2681-2688.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Di Virgilio F., Meyer B. C., Greenberg S., Silverstein S. C. Fc receptor-mediated phagocytosis occurs in macrophages at exceedingly low cytosolic Ca2+ levels. J Cell Biol. 1988 Mar;106(3):657–666. doi: 10.1083/jcb.106.3.657. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Griffin F. M., Jr, Griffin J. A., Leider J. E., Silverstein S. C. Studies on the mechanism of phagocytosis. I. Requirements for circumferential attachment of particle-bound ligands to specific receptors on the macrophage plasma membrane. J Exp Med. 1975 Nov 1;142(5):1263–1282. doi: 10.1084/jem.142.5.1263. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Grynkiewicz G., Poenie M., Tsien R. Y. A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem. 1985 Mar 25;260(6):3440–3450. [PubMed] [Google Scholar]
  7. 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]
  8. Hazama A., Yada T., Okada Y. HeLa cells have histamine H1-receptors which mediate activation of the K+ conductance. Biochim Biophys Acta. 1985 May 30;845(2):249–253. doi: 10.1016/0167-4889(85)90183-1. [DOI] [PubMed] [Google Scholar]
  9. Janmey P. A., Stossel T. P. Modulation of gelsolin function by phosphatidylinositol 4,5-bisphosphate. Nature. 1987 Jan 22;325(6102):362–364. doi: 10.1038/325362a0. [DOI] [PubMed] [Google Scholar]
  10. Kruskal B. A., Maxfield F. R. Cytosolic free calcium increases before and oscillates during frustrated phagocytosis in macrophages. J Cell Biol. 1987 Dec;105(6 Pt 1):2685–2693. doi: 10.1083/jcb.105.6.2685. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Labigne-Roussel A. F., Lark D., Schoolnik G., Falkow S. Cloning and expression of an afimbrial adhesin (AFA-I) responsible for P blood group-independent, mannose-resistant hemagglutination from a pyelonephritic Escherichia coli strain. Infect Immun. 1984 Oct;46(1):251–259. doi: 10.1128/iai.46.1.251-259.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. Lassing I., Lindberg U. Specific interaction between phosphatidylinositol 4,5-bisphosphate and profilactin. Nature. 1985 Apr 4;314(6010):472–474. doi: 10.1038/314472a0. [DOI] [PubMed] [Google Scholar]
  14. Lew D. P., Andersson T., Hed J., Di Virgilio F., Pozzan T., Stendahl O. Ca2+-dependent and Ca2+-independent phagocytosis in human neutrophils. Nature. 1985 Jun 6;315(6019):509–511. doi: 10.1038/315509a0. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. Maurelli A. T., Sansonetti P. J. Genetic determinants of Shigella pathogenicity. Annu Rev Microbiol. 1988;42:127–150. doi: 10.1146/annurev.mi.42.100188.001015. [DOI] [PubMed] [Google Scholar]
  17. McNeil P. L., Swanson J. A., Wright S. D., Silverstein S. C., Taylor D. L. Fc-receptor-mediated phagocytosis occurs in macrophages without an increase in average [Ca++]i. J Cell Biol. 1986 May;102(5):1586–1592. doi: 10.1083/jcb.102.5.1586. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Moulder J. W. Comparative biology of intracellular parasitism. Microbiol Rev. 1985 Sep;49(3):298–337. doi: 10.1128/mr.49.3.298-337.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Murata T., Sullivan J. A., Sawyer D. W., Mandell G. L. Influence of type and opsonization of ingested particle on intracellular free calcium distribution and superoxide production by human neutrophils. Infect Immun. 1987 Aug;55(8):1784–1791. doi: 10.1128/iai.55.8.1784-1791.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Osada Y., Une T., Ikeuchi T., Ogawa H. Effect of calcium on the cell infectivity of virulent Shigella flexneri 2a. Jpn J Microbiol. 1974 Jul;18(4):321–326. doi: 10.1111/j.1348-0421.1974.tb00816.x. [DOI] [PubMed] [Google Scholar]
  21. Painter R. G., Whisenand J., McIntosh A. T. Effects of cytochalasin B on actin and myosin association with particle binding sites in mouse macrophages: implications with regard to the mechanism of action of the cytochalasins. J Cell Biol. 1981 Nov;91(2 Pt 1):373–384. doi: 10.1083/jcb.91.2.373. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Pollard T. D., Cooper J. A. Actin and actin-binding proteins. A critical evaluation of mechanisms and functions. Annu Rev Biochem. 1986;55:987–1035. doi: 10.1146/annurev.bi.55.070186.005011. [DOI] [PubMed] [Google Scholar]
  23. Renard D., Poggioli J., Berthon B., Claret M. How far does phospholipase C activity depend on the cell calcium concentration? A study in intact cells. Biochem J. 1987 Apr 15;243(2):391–398. doi: 10.1042/bj2430391. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. 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]
  25. Sansonetti P. J., Ryter A., Clerc P., Maurelli A. T., Mounier J. Multiplication of Shigella flexneri within HeLa cells: lysis of the phagocytic vacuole and plasmid-mediated contact hemolysis. Infect Immun. 1986 Feb;51(2):461–469. doi: 10.1128/iai.51.2.461-469.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Sauvé R., Simoneau C., Parent L., Monette R., Roy G. Oscillatory activation of calcium-dependent potassium channels in HeLa cells induced by histamine H1 receptor stimulation: a single-channel study. J Membr Biol. 1987;96(3):199–208. doi: 10.1007/BF01869302. [DOI] [PubMed] [Google Scholar]
  27. Sawyer D. W., Sullivan J. A., Mandell G. L. Intracellular free calcium localization in neutrophils during phagocytosis. Science. 1985 Nov 8;230(4726):663–666. doi: 10.1126/science.4048951. [DOI] [PubMed] [Google Scholar]
  28. Sheterline P., Rickard J. E., Richards R. C. Fc receptor-directed phagocytic stimuli induce transient actin assembly at an early stage of phagocytosis in neutrophil leukocytes. Eur J Cell Biol. 1984 May;34(1):80–87. [PubMed] [Google Scholar]
  29. Steinberg T. H., Newman A. S., Swanson J. A., Silverstein S. C. Macrophages possess probenecid-inhibitable organic anion transporters that remove fluorescent dyes from the cytoplasmic matrix. J Cell Biol. 1987 Dec;105(6 Pt 1):2695–2702. doi: 10.1083/jcb.105.6.2695. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Stendahl O. I., Hartwig J. H., Brotschi E. A., Stossel T. P. Distribution of actin-binding protein and myosin in macrophages during spreading and phagocytosis. J Cell Biol. 1980 Feb;84(2):215–224. doi: 10.1083/jcb.84.2.215. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Stewart D. I., Crawford N. Redistribution of membrane-bound and cytosolic action in rabbit polymorphonuclear leucocytes during phagocytosis. Biochem J. 1985 Feb 1;225(3):807–814. doi: 10.1042/bj2250807. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Tsien R. Y. New calcium indicators and buffers with high selectivity against magnesium and protons: design, synthesis, and properties of prototype structures. Biochemistry. 1980 May 27;19(11):2396–2404. doi: 10.1021/bi00552a018. [DOI] [PubMed] [Google Scholar]
  33. Tsien R. Y., Pozzan T., Rink T. J. Calcium homeostasis in intact lymphocytes: cytoplasmic free calcium monitored with a new, intracellularly trapped fluorescent indicator. J Cell Biol. 1982 Aug;94(2):325–334. doi: 10.1083/jcb.94.2.325. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES