1887

Abstract

Summary

The behaviour of lysosomal enzymes in poliovirus-infected HeLa S3 cells and vesicular stomatitis virus (VSV)-infected L cells was investigated both biochemically using enzyme assays, and histochemically using acid phosphatase dependent staining. The presence of the enzyme was shown histochemically under the light microscope by its reaction with naphthole-AS-BI-phosphate and a coupling reaction with diazotized pararosaniline. The light absorption of this stain in infected and uninfected cells was measured on a Universal Micro Spectrophotometer (UMSP-1) and recorded on-line as gray value cell images in a PDP-12 computer. These scanned images were analyzed by FORTRAN programs on a UNIVAC1108. The histochemically obtained distributions of the lysosomal enzyme are comparable to the results of the biochemical analysis. Lysosomes of poliovirus-infected cells displayed a release of lysosomal enzymes into the cytoplasm starting at 3 h after infection; VSV infection did not produce this type of effect. This investigation shows that it is possible to extract and demonstrate specific virus dependent changes using computer-aided cytophotometric techniques.

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/content/journal/jgv/10.1099/0022-1317-25-3-359
1974-12-01
2024-04-20
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References

  1. Allison A. C., Sandelin K. 1963; Activation of lysosomal enzymes in virus-infected cells and its possible relationship to cytophatic effect. Journal of Experimental Medicine 117:879–887
    [Google Scholar]
  2. Allison A. C., Malucci L. 1965; Histochemical studies of lysosomes and lysosomal enzymes in virus-infected cell cultures. Journal of Experimental Medicine 121:463–476
    [Google Scholar]
  3. Amako K., Dales S. 1967; Cytopathology of Mengo virus infection. II. Proliferation of membranous cisternae. Virology 32:201–215
    [Google Scholar]
  4. Aus H. M., Meulen V. ter, Kaekell M., Scholz W., Koschel K. 1974; Techniques applicable to computer-aided cytophotometry in virology. The Journal of Histochemistry and Cytochemistry 22:577–582
    [Google Scholar]
  5. Barka T., Anderson P. J. 1962; Histochemical methods for acid phosphatase using hexazonium pararo-saniline as coupler. Journal of Histochemistry and Cytochemistry 10:741–753
    [Google Scholar]
  6. Blackman K. E., Bubel H. C. 1969; Poliovirus induced cellular injury. Journal of Virology 4:203–208
    [Google Scholar]
  7. Bray G. A. 1960; A simple efficient liquid scintillator for counting aqueous solutions in a liquid scintillation counter. Analytical Biochemistry 1:279–285
    [Google Scholar]
  8. Casparsson T., Lomakka G., Svensson G., Saftström R. 1955; A versatile ultramicrospectograph for multiple-line and surface scanning high resolution measurements employing automatized data analysis. Experimental Cell Research (supplement) 340–51
    [Google Scholar]
  9. Defendi V. 1962; Cytopathology of virus infection. Federation Proceedings of the American Societies for Experimental Biology 21:1113–1117
    [Google Scholar]
  10. Dulbecco R. 1952; Production of plaques in monolayer tissue cultures by single particles of an animal virus. Proceedings of the National Academy of Science of the United States of America 38:747–752
    [Google Scholar]
  11. Flanagan J. F. 1966; Hydrolytic enzymes in KB cells infected with poliovirus and herpes simplex virus. Journal of Bacteriology 91:789–797
    [Google Scholar]
  12. Guskey L. E., Smith P. C., Wolff D. A. 1970; Patterns of cytopathology and lysosomal enzyme release in polio-infected HEp-2 cells treated with either 2-(α-hydroxybenzyl)-benzimidazole or guanidine HC1. Journal of General Virology 6:151–161
    [Google Scholar]
  13. Horak L., Jungwirth C., Bodo G. 1971; Poxvirus-specific cytopathic effect in interferon-treated L-cells. Virology 45:456–462
    [Google Scholar]
  14. Killington R. A., Lee D., Stott E. J., Osborne J. A. 1974; Studies on the lysosomes of L 132 cells infected with either rhinovirus type 2 or poliovirus type 1. Journal of General Virology 22:303–307
    [Google Scholar]
  15. Koschel K. 1971; Release of 51Chromium from labelled HeLa cells after infection by poliovirus. Zeitschrift für Naturforschung 26b:929–933
    [Google Scholar]
  16. Lowry H. O., Rosebrough N. J., Farr A. L., Randall R. J. 1951; Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry 193:265–275
    [Google Scholar]
  17. Macieira-Coelho A., Fernandes M. V., Mellman W. J. 1965; Cortisone on in vitro infection. II. Effect on distribution of cytopathic effect and acid phosphatase arrangement in human cells infected with poliovirus. Proceedings of the Society for Experimental Biology and Medicine 119:631–635
    [Google Scholar]
  18. Mosser A. G., Caliguiri L. A., Tamm I. 1972; Incorporation of lipid precursors into membranes of poliovirus-infected HeLa cells. Virology 47:39–47
    [Google Scholar]
  19. Plagemann P. G. W., Cleveland P. H., Shea M. A. 1970; Effect of Mengovirus replication on choline metabolism and membrane formation in Novikoff hepatoma cells. Journal of Virology 6:800–812
    [Google Scholar]
  20. Wolff D. A., Bubel H. C. 1964; The disposition of lysosomal enzymes as related to specific viral cytopathic effects. Virology 24:502–505
    [Google Scholar]
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