Archives of Virology

, Volume 92, Issue 3–4, pp 273–291

An electron and immunoelectron microscopic study of dengue-2 virus infection of cultured mosquito cells: Maturation events

  • T. Hase
  • P. L. Summers
  • K. H. Eckels
  • W. B. Baze
Original Papers


The maturation process of dengue-2 virus in C6/36 mosquito cells was studied by electron microscopy at 12, 16, 24, 48, and 78 hours postinoculation (p.i.) and by immunoelectron microscopy at 48 and 78 hours p.i. Maturing virions appeared within cytoplasmic vacuoles and on the surface of infected cells from 24 hours p.i. onward in close topographical relationship to the dense particles that occurred concurrently in the cytoplasm. The dense particles measured 25 to 35 nm in diameter; the mature virions measured 50 to 55 nm in diameter, with a dense core measuring 30 to 35 nm in diameter covered by a 10 nm-thick membrane envelope. The morphological observations indicated that the dense particles were dengue nucleocapsids assembled in the cytoplasm and that they apparently budded into the vacuolar lumens and the extracellular space at the vacuolar and plasma membranes, acquiring membrane envelopes and becoming mature virions in the process. The virions that budded into the vacuolar lumens were released extracellularly by exocytosis. In the samples tested with dengue-2 polyclonal antibodies, intense immunostaining occurred at the sites of virus budding on the cell surface; host cell membrane and cytoplasm adjacent to the budding virions stained less intensely. In the samples tested with a dengue-2 monoclonal antibody specific for the envelope glycoprotein, budding virions stained rather exclusively, with no staining occurring in adjacent host membrane or cytoplasm.


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  1. 1.
    Achison NH, Tamm I (1967) Replication of Semliki Forest virus: an electron microscopic study. Virology 32: 128–143Google Scholar
  2. 2.
    Brown DT (1980) The assembly of alphaviruses. In:Schlesinger RW (ed) The togaviruses. Biology, structure, replication. Academic Press, New York, pp 437–501Google Scholar
  3. 3.
    Cardiff RD, Lund JK (1976) Distribution of dengue-2 antigens by electron immunochemistry. Infect Immun 13: 1699–1709Google Scholar
  4. 4.
    Cardiff RD, Russ SB, Brandt WE, Russell PK (1973) Cytological localization of dengue-2 antigens: an immunological study with ultrastructural correlation. Infect Immun 7: 809–816Google Scholar
  5. 5.
    Catanzaro PJ, Brandt WE, Hogrefe WR, Russell PK (1974) Detection of dengue cell-surface antigens by peroxidase-labeled antibodies and immune cytolysis. Infect Immun 10: 381–388Google Scholar
  6. 6.
    Demsey A, Steere RR, Brandt WE, Veltri BJ (1974) Morphology and development of dengue-2 virus employing the freeze-fracture and thin-section technique. J Ultrastruct Res 46: 103–116Google Scholar
  7. 7.
    Eckels KH, Brandt WE, Harrison VR, McCown JM, Russell PK (1976) Isolation of a temperature-sensitive dengue-2 virus under conditions suitable for vaccine development. Infect Immun 14: 1221–1227Google Scholar
  8. 8.
    Fenner F, McAuslan BR, Mims CA, Sambrook F, White DO (1974) The biology of animal viruses, 2nd edn. Academic Press, New York, pp 24–25Google Scholar
  9. 9.
    Filshie BK, Rehacek J (1968) Studies of the morphology of Murray Valley encephalitis and Japanese encephalitis viruses growing in cultured mosquito cells. Virology 34: 435–443Google Scholar
  10. 10.
    Igarashi A (1978) Isolation of a Singh'sAedes albopictus cell clone sensitive to dengue and Chikungunya viruses. J Gen Virol 40: 531–544Google Scholar
  11. 11.
    Ko KK (1976) Development of dengue virus type 4 in BHK 21 cells. Biken J 19: 43–51Google Scholar
  12. 12.
    Ko KK, Igarashi A, Fukai K (1979) Electron microscopic observation onAedes albopictus cells infected with dengue viruses. Arch Virol 62: 41–52Google Scholar
  13. 13.
    Leary K, Blair CD (1980) Sequential events in the morphogenesis of Japanese encephalitis virus. J Ultrastruc Res 72: 123–129Google Scholar
  14. 14.
    Matsumura T, Shirai K, Sashikata T, Hotta S (1977) Morphogenesis of dengue-1 virus in cultures of a human leukemic leukocyte line (J-111). Microbiol Immunol 21: 329–334Google Scholar
  15. 15.
    Matsumura T, Stoller V, Schlesinger RW (1971) Studies on the nature of dengue viruses. V. Structure and development of dengue virus in vero cells. Virology 46: 344–355Google Scholar
  16. 16.
    Murphy FA (1980) Togavirus morphology and morphogenesis. In:Schlesinger RW (ed) The togaviruses. Biology, structure, replication. Academic Press, New York, pp 241–315Google Scholar
  17. 17.
    Ota E (1965) Electron microscopic study of the development of Japanese encephalitis virus in porcine kidney stable (PS) cells. Virology 25: 372–378Google Scholar
  18. 18.
    Shapiro D, Brandt WE, Russell PK (1972) Changes involving viral membrane glycoprotein during morphogenesis of group B arboviruses. Virology 50: 906–911Google Scholar
  19. 19.
    Singh KRP (1967) Cell cultures derived from larvae ofAedes albopictus (Skuse) andAedes aegypti (L.). Curr Sci 36: 506–508Google Scholar
  20. 20.
    Sriurairatna S, Bhamarapravati N (1977) Replication of dengue-2 virus inAedes albopictus mosquitoes. Am J Trop Med Hyg 26: 1199–1205Google Scholar
  21. 21.
    Sriurairatna S, Bhamarapravati N, Onishi S (1974) Filamentous structures in dengue type 3 virus infected mouse neurons. Biken J 17: 183–191Google Scholar
  22. 22.
    Sriurairatna S, Bhamarapravati N, Phalavadhtna O (1973) Dengue virus infection of mice: morphology and morphogenesis of dengue type-2 virus in suckling mouse neurons. Infect Immun 8: 1017–1028Google Scholar
  23. 23.
    Stohlman SA, Wisseman CL, Eyler OR, Silverman DJ (1975) Dengue virus-induced modification of host cell membranes. J Virol 16: 1017–1026Google Scholar
  24. 24.
    Stoller V (1980) Togaviruses in cultured arthropod cells. In:Schlesinger RW (ed) The togaviruses. Biology, structure, replication. Academic Press, New York, pp 583–621Google Scholar
  25. 25.
    Westaway EG (1980) Replication of flaviviruses. In:Schlesinger RW (ed) The togaviruses. Biology, structure, replication. Academic Press, New York, pp 531–581Google Scholar
  26. 26.
    Whitfield SG, Murphy FA, Sudia WD (1973) St. Louis encephalitis virus: an ultra-structural study of infection in a mosquito vector. Virology 56: 70–87Google Scholar

Copyright information

© Springer-Verlag 1987

Authors and Affiliations

  • T. Hase
    • 1
  • P. L. Summers
    • 2
  • K. H. Eckels
    • 2
  • W. B. Baze
    • 1
  1. 1.Department of Ultrastructural StudiesWalter Reed Army Institute of ResearchWashington, D.C.U.S.A.
  2. 2.Department of Biologies ResearchWalter Reed Army Institute of ResearchWashington, D.C.U.S.A.

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