1887

Abstract

Summary

Cytotoxic activity of culture supernates of is manifested by vacuolation of mammalian cells The formation and maturation of toxin-induced vacuoles in HeLa cells has been studied to examine the possibility that they are autophagosomal in nature. Observation by light microscopy revealed that vacuoles originate in a perinuclear position, increasing in number and size until cell degeneration and lysis occur after 48 h. Ultrastructural study of mature vacuoles indicated the presence of a bounding membrane with contents consisting of degenerate cytoplasmic components and acid phosphatase activity. Confocal fluorescence imaging demonstrated acridine orange accumulation in the vacuoles of toxin-treated cells, indicating an acidic intravacuolar pH. These features are characteristic of autophagosomes. In addition, the size of vacuoles in living, acridine orange-stained cells tended to be inversely proportional to fluorescence intensity. Fluid phase endocytic markers were observed only rarely within nascent vacuoles. Over the succeeding 24 h, labelling of most vacuoles with these dyes was observed. This, along with the observation of intravacuolar acid phosphatase activity, provides evidence that vacuoles communicate at some point during their development with endocytically derived compartments. These observations provide direct evidence for an autophagic origin of these structures.

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1992-12-01
2024-03-28
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References

  1. Blaser M. J. Gastric Campylobacter-like organisms, gastritis, and peptic ulcer disease. Gastroenterology 1987; 93:371–383
    [Google Scholar]
  2. Goodwin C. S., Armstrong J. A., Marshall B. J. Campylobacter pyloridis, gastritis and peptic ulceration. J Clin Pathol 1986; 39:353–365
    [Google Scholar]
  3. Marshall B. J., Armstrong J. A., McGechie D. B., Glancy R. J. Attempt to fulfil Koch’s postulates for pyloric Campylobacter. MedJAust 1985; 142:436–439
    [Google Scholar]
  4. Leunk R. D., Johnson P. T., David B. C., Kraft W. G., Morgan D. R. Cytotoxic activity in broth-culture filtrates of Campylobacter pylori. J Med Microbiol 1988; 26:93–99
    [Google Scholar]
  5. Eaton K. A., Morgan D. R., Krakowka S. Campylobacter pylori virulence factors in gnotobiotic piglets. Infect Immun 1989; 57:1119–125
    [Google Scholar]
  6. Figura N., Guglielmetti P., Rossolini A. Cytotoxin production by Campylobacter pylori strains isolated from patients with peptic ulcers and from patients with chronic gastritis only. J Clin Microbiol 1989; 27:225–226
    [Google Scholar]
  7. Leunk R. D., Ferguson M. A., Morgan D. R., Low D. E., Simor A. E. Antibody to cytotoxin in infection by Helicobacter pylori. J Clin Microbiol 1990; 28:1181–1184
    [Google Scholar]
  8. Tricottet V., Bruneval P., Vire O. Campylobacter-like organisms and surface epitheloium abnormalities in active, chronic gastritis in humans: an ultrastructural study. Ultrastruct Pathol 1986; 10:113–122
    [Google Scholar]
  9. Catrenich C. E., Chestnut M. H. Characterization of the vacuoles induced by the cytotoxin of Helicobacter pylori. Abstracts of the 91st Annual Meeting of the American Society for Microbiology 1991 Abstract B-212 61
    [Google Scholar]
  10. Chestnut M. H., Catrenich C. E. Vacuoles induced in mammalian cells by Helicobacter pylori are acidic organelles. Proceedings of the Xllth International Congress for Electron Microscopy 1990168 (Abstract)
    [Google Scholar]
  11. Cover T. L., Puryear W., Perez-Perez G. I., Blaser M. J. Effect of urease on HeLa cell vacuolation induced by Helicobacter pylori cytotoxin. Infect Immun 1991; 59:1264–1270
    [Google Scholar]
  12. Dunn W. A. Studies on the mechanisms of autophagy: formation of the autophagic vacuole. J Cell Biol 1990; 110:1923–1933
    [Google Scholar]
  13. Dunn W. A. Studies on the mechanisms of autophagy: maturation of the autophagic vacuole. J Cell Biol 1990; 110:1935–1945
    [Google Scholar]
  14. Arstila A. U., Trump B. F. Studies on cellular autophagocytosis. The formation of autophagic vacuoles in the liver after glucagon administration. Am J Pathol 1968; 53:687–733
    [Google Scholar]
  15. Hubbard A. L. Endocytosis. Curr Opin Cell Biol 1989; 1:675–683
    [Google Scholar]
  16. Kornfeld S. Trafficking of lysosomal enzymes. FASEB J 1987; 1:462–468
    [Google Scholar]
  17. DeDuve C., Wattiaux R. Functions of lysosomes. Annu Rev Physiol 1966; 28:435–492
    [Google Scholar]
  18. Matteoni R., Kreis T. E. Translocation and clustering of endo-somes and lysosomes depends on microtubules. J Cell Biol 1987; 105:1253–1265
    [Google Scholar]
  19. Plant A. L., Benson D. M., Smith L. C. Cellular uptake and intracellular localization of benzo (a)pyrene by digital fluorescence imaging microscopy. J Cell Biol 1985; 100:1295–1308
    [Google Scholar]
  20. Griffiths G. W. Transport of glial cell acid phosphatase by endoplasmic reticulum into damaged axons. J Cell Sci 1979; 36:361–389
    [Google Scholar]
  21. Griffiths G., Quinn P., Warren G. Dissection of the golgi complex. 1. Monensin inhibits the transport of viral membrane proteins from medial to trans golgi cistemae in baby hamster kidney cells infected with Semliki forest virus. J Cell Biol 1983; 96:835–850
    [Google Scholar]
  22. Spurr A. R. A low-viscosity expoxy resin embedding medium for electron microscopy. J Ultrastruct Res 1969; 26:31–43
    [Google Scholar]
  23. Mitchener J. S., Shelburne J. D., Bradford W. D., Hawkins H. K. Cellular autophagocytosis induced by deprivation of serum and amino acids in HeLa cells. Am J Pathol 1976; 83:485–498
    [Google Scholar]
  24. Woods D. E., Hwang W. S., Shahrabadi M. S., Que J. U. Alteration of pulmonary structure by Pseudomonas aeruginosa exoenzyme, S. J Med Microbiol 1988; 26:133–141
    [Google Scholar]
  25. Tsien R. Y. Fluorescent indicators of ion concentrations. In Taylor D. L., Wang Y. -L. (eds) Methods in cell biology vol 30 Academic Press, Inc; 1989127–156
    [Google Scholar]
  26. Cain C. C., Murphy R. F. Growth inhibition of 3T3 fibroblasts by lysosomotropic amines: correlation with effects on intravesicular pH but not vacuolation. J Cell Physiol 1986; 129:65–70
    [Google Scholar]
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