A Quantitative Ultrastructural Study of the Development of Bluetongue Virus in Madin-Darby Bovine Kidney Cells Free

Abstract

SUMMARY

Samples of Madin-Darby bovine kidney cells were taken for electron microscopy at various times after infection with bluetongue virus. A quantitative electron microscopic cell sampling technique was used in observing ultrathin sections of the cell population, and various morphological features were tabulated. The appearance of virus within phagocytic vesicles and lysosomes, and the distension of the rough endoplasmic reticulum, were early ultrastructural changes resulting from the virus infection. Granular inclusions which were usually juxtanuclear and tubular structures were other features observed later in cells infected with bluetongue virus. Progeny virus particles were seen within granular inclusions, amidst tubular structures, and within cytoplasmic vesicles. There was agreement between the infectivity data from plaque counts of cell-associated virus and direct electron microscopic counts of cells containing intracellular virus, granular inclusions, and cytoplasmic tubular structures. The large tubules associated with bluetongue infection had a mean outer diameter of 47.2 nm., more than twice the size of classic cellular microtubules. The mean diameter of bluetongue virus measured in ultrathin sections and negatively stained preparations was 63 nm. Even at late stages of infection, small numbers of intracellular virus particles were observed with the electron microscope. This agreed with the low titres obtained from plaque counts of cell-associated virus. Bluetongue virus did not, therefore, accumulate in the cell and undergo a burst-like release. Instead, it appeared to be extruded from the cell as it was made. There was no evidence for virus release by budding from the plasma membrane, nor for the presence of an envelope around complete particles.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-13-2-229
1971-11-01
2024-03-28
Loading full text...

Full text loading...

/deliver/fulltext/jgv/13/2/JV0130020229.html?itemId=/content/journal/jgv/10.1099/0022-1317-13-2-229&mimeType=html&fmt=ahah

References

  1. Anderson N., Doane F. W. 1966; An electron-microscope study of reovirus type 2 in L cells. Journal of Pathology and Bacteriology 92:433
    [Google Scholar]
  2. Bowne J. O., Jochim M. M. 1967; Cytopathologic changes and development of inclusion bodies in cultured cells infected with bluetongue virus. American Journal of Veterinary Research 28:1091
    [Google Scholar]
  3. Bowne J. G., Tones R. H. 1966; Observations on bluetongue virus in the salivary glands of an insect vector, Culicoides variipennis . Virology 30:127
    [Google Scholar]
  4. Bowne J. G., Ritchie A. E. 1970; Some morphological features of bluetongue virus. Virology 40:903
    [Google Scholar]
  5. Dales S. 1963; Association between the spindle apparatus and reovirus. Proceedings of the National Academy of Sciences of the United States of America 50:268
    [Google Scholar]
  6. Dales S., Gomatos P. J., Hsu K. C. 1965; The uptake and development of reovirus in strain L cells followed with labelled viral RNA and ferritin-antibody conjugates. Virology 25:193
    [Google Scholar]
  7. Dulbecco R., Vogt M. 1954; One-step growth curve of western equine encephalomyelitis virus in chicken embryo cells grown in vitro and analysis of virus yields from single cells. Journal of Experimental Medicine 99:183
    [Google Scholar]
  8. Els H. J., Verwoerd D. W. 1969; Morphology of bluetongue virus. Virology 38:213
    [Google Scholar]
  9. Foster N. M., Jones R. H., Mccrory B. R. 1963; Preliminary investigations on the insect transmission of bluetongue in sheep. American Journal of Veterinary Research 24:1195
    [Google Scholar]
  10. Gomatos P. J., Tamm I., Dales S., Frankun R. M. 1962; Reovirus type 3: Physical characteristics and interaction with L cells. Virology 17:441
    [Google Scholar]
  11. Harford C. H., Hamlin A., Middelkamp J. N., Briggs D. D. Jun 1962; Electron microscopic examination of cells infected with reovirus. Journal of Laboratory and Clinical Medicine 60:179
    [Google Scholar]
  12. Hassan S. A., Rabin E. R., Melnick J. L. 1965; Reovirus myocarditis in mice: An electron-microscopic, immunofluorescent and virus assay study. Experimental Molecular Pathology 4:66
    [Google Scholar]
  13. Jenson A. B., Rabin E. R., Phillips C. A., Melnick J. L. 1965; Reovirus encephalitis in newborn mice: An electron-microscopic and virus assay study. American Journal of Pathology 47:223
    [Google Scholar]
  14. Jochim M. M., Jones R. H. 1966; Multiplication of bluetongue virus in Culicoides variipennis following artificial infection. American Journal of Epidemiology 84:241
    [Google Scholar]
  15. Lecatsas G. 1968a; Electron microscopic study of the formation of bluetongue virus. Onderstepoort Journal of Veterinary Research 35:139
    [Google Scholar]
  16. Lecatsas G. 1968b; Electron microscopic studies on reovirus type 1 in BHK. 21 cells. Onderstepoort Journal of Veterinary Research 35:151
    [Google Scholar]
  17. Loh P. C., Hohl H. R., Soergel M. 1965; Fine structure of reovirus type 2. Journal of Bacteriology 89:1140
    [Google Scholar]
  18. Madin S. H., Darby N. B. 1958; Established kidney cell lines of normal adult bovine and ovine origin. Proceedings of the Society for Experimental Biology and Medicine 98:574
    [Google Scholar]
  19. Mollenhauer H. H. 1964; Plastic embedding mixtures for use in electron microscopy. Stain Technology 39:111
    [Google Scholar]
  20. Owen N. C., Munz E. K. 1966; Observations on a strain of bluetongue virus by electron microscopy. Onderstepoort Journal of Veterinary Research 33:9
    [Google Scholar]
  21. Reynolds E. S. 1963; The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. Journal of Cell Biology 17:208
    [Google Scholar]
  22. Rhim J. S., Melnick J. L. 1961; Plaque formation by reoviruses. Virology 15:80
    [Google Scholar]
  23. Silverstein S. C., Dales S. 1968; The penetration of reovirus RNA and initiation of its genetic function in L-strain fibroblasts. Journal of Cell Biology 36:197
    [Google Scholar]
  24. Slautterback D. B. 1963; Cytoplasmic microtubules, L Hydra. Journal of Cell Biology 18:367
    [Google Scholar]
  25. Studdert M. J. 1965; Sensitivity of bluetongue virus to ether and sodium desoxycholate. Proceedings of the Society for Experimental Biology and Medicine 118:1006
    [Google Scholar]
  26. Studdert M. J., Pangborn J., Addison R. B. 1966; Bluetongue virus structure. Virology 29:480
    [Google Scholar]
  27. Svehag S. E., Leendertsen L., Gorham J. R. 1966; Sensitivity of bluetongue virus to lipid solvents, trypsin, and pH changes and its serological relationship to arboviruses. Journal of Hygiene, Cambridge 64:339
    [Google Scholar]
  28. Tournier P., Plissier M. 1960; Le développement intracellulaire du réovirus observé au microscope électronique. La Presse Médicale 68:683
    [Google Scholar]
  29. Vasquez C., Tournier P. 1962; The morphology of reovirus. Virology 17:503
    [Google Scholar]
  30. Verwoerd D. W. 1969; Purification and characterization of bluetongue virus. Virology 38203
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-13-2-229
Loading
/content/journal/jgv/10.1099/0022-1317-13-2-229
Loading

Data & Media loading...

Most cited Most Cited RSS feed