Skip to main content
Infection and Immunity logoLink to Infection and Immunity
. 1973 May;7(5):809–816. doi: 10.1128/iai.7.5.809-816.1973

Cytological Localization of Dengue-2 Antigens: an Immunological Study with Ultrastructural Correlation

R D Cardiff a,1, S B Russ a, W E Brandt a, P K Russell a
PMCID: PMC422765  PMID: 4202963

Abstract

Immunofluorescence and electron microscopy were used to study the development and cytological distribution of dengue-2 virion and nonvirion antigens in monkey kidney cells. The type and combination of fixatives were found to affect the intensity of fluorescence. Paraformaldehyde fixation alone resulted in a low level of fluorescence, but an additional fixation step with ethanol or acetone resulted in maximum staining. No fluorescence was obtained after fixation with glutaraldehyde alone or in combination with other solvents. Methanol caused selective ablation of fluorescence by antibody to purified virions. Dengue-specific fluorescence that was most intense in the perinuclear area radiated in a granular pattern of decreasing intensity into the cytoplasm. The perinuclear fluorescence was associated with nonvirion antigens and the cytoplasmic fluorescence was associated with virion antigens. Electron micrographs of infected cells revealed vesicular bodies with reticular electron-dense centers, clusters of structurally complete virions deep within the endoplasmic reticulum, and single virus particles in vacuoles near the periphery of the cell.

Full text

PDF
809

Images in this article

Selected References

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

  1. Acheson N. H., Tamm I. Replication of Semliki Forest virus: an electron microscopic study. Virology. 1967 May;32(1):128–143. doi: 10.1016/0042-6822(67)90261-9. [DOI] [PubMed] [Google Scholar]
  2. BHAMARAPRAVATI N., HALSTEAD S. B., SOOKAVACHANA P., BOONYAPAKNAVIK V. STUDIES ON DENGUE VIRUS INFECTION. 1. IMMUNOFLUORESCENT LOCALIZATION OF VIRUS IN MOUSE TISSUE. Arch Pathol. 1964 May;77:538–543. [PubMed] [Google Scholar]
  3. Brandt W. E., Cardiff R. D., Russell P. K. Dengue virions and antigens in brain and serum of infected mice. J Virol. 1970 Oct;6(4):500–506. doi: 10.1128/jvi.6.4.500-506.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brandt W. E., Chiewslip D., Harris D. L., Russell P. K. Partial purification and characterization of a dengue virus soluble complement-fixing antigen. J Immunol. 1970 Dec;105(6):1565–1568. [PubMed] [Google Scholar]
  5. Bykovsky A. F., Yershov F. I., Zhdanov V. M. Morphogenesis of Venezuelan equine encephalomyelitis virus. J Virol. 1969 Oct;4(4):496–504. doi: 10.1128/jvi.4.4.496-504.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cardiff R. D., Brandt W. E., McCloud T. G., Shapiro D., Russell P. K. Immunological and biophysical separation of dengue-2 antigens. J Virol. 1971 Jan;7(1):15–23. doi: 10.1128/jvi.7.1.15-23.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Eldadah A. H., Nathanson N. Pathogenesis of West Nile Virus encepahlitis in mice and rats. II. Virus multiplication, evolution of immunofluorescence, and development of histological lesions in the brain. Am J Epidemiol. 1967 Nov;86(3):776–790. doi: 10.1093/oxfordjournals.aje.a120786. [DOI] [PubMed] [Google Scholar]
  8. Filshie B. K., Rehacek J. Studies of the morphology of Murray Valley encephalitis and Japanese encephalitis viruses growing in cultured mosquito cells. Virology. 1968 Mar;34(3):435–443. doi: 10.1016/0042-6822(68)90063-9. [DOI] [PubMed] [Google Scholar]
  9. Grimley P. M., Berezesky I. K., Friedman R. M. Cytoplasmic structures associated with an arbovirus infection: loci of viral ribonucleic acid synthesis. J Virol. 1968 Nov;2(11):1326–1338. doi: 10.1128/jvi.2.11.1326-1338.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hahon N. Cell surface antigen induced by venezuelan equine encephalomyelitis virus. Infect Immun. 1970 Dec;2(6):713–715. doi: 10.21236/ad0872045. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Matsumura T., Stollar V., Schlesinger R. W. Studies on the nature of dengue viruses. V. Structure and development of dengue virus in Vero cells. Virology. 1971 Nov;46(2):344–355. doi: 10.1016/0042-6822(71)90036-5. [DOI] [PubMed] [Google Scholar]
  12. McCloud T. G., Brandt W. E., Russell P. K. Molecular size and charge relationships of the soluble complement-fixing antigens of dengue viruses. Virology. 1970 Jul;41(3):569–572. doi: 10.1016/0042-6822(70)90180-7. [DOI] [PubMed] [Google Scholar]
  13. Murphy F. A., Harrison A. K., Gary G. W., Jr, Whitfield S. G., Forrester F. T. St. Louis encephalitis virus infection in mice. Electron microscopic studies of central nervous system. Lab Invest. 1968 Dec;19(6):652–662. [PubMed] [Google Scholar]
  14. OTA Z. ELECTRON MICROSCOPE STUDY OF THE DEVELOPMENT OF JAPANESE B ENCEPHALITIS VIRUS IN PORCINE KIDNEY STABLE (PS) CELLS. Virology. 1965 Mar;25:372–378. doi: 10.1016/0042-6822(65)90057-7. [DOI] [PubMed] [Google Scholar]
  15. Robb J. A., Martin R. G. Genetic analysis of simian virus 40. I. Description of microtitration and replica-plating techniques for virus. Virology. 1970 Aug;41(4):751–760. doi: 10.1016/0042-6822(70)90439-3. [DOI] [PubMed] [Google Scholar]
  16. Russell P. K., Chiewsilp D., Brandt W. E. Immunoprecipitation analysis of soluble complement-fixing antigens of dengue viruses. J Immunol. 1970 Oct;105(4):838–845. [PubMed] [Google Scholar]
  17. Russell P. K., Nisalak A., Sukhavachana P., Vivona S. A plaque reduction test for dengue virus neutralizing antibodies. J Immunol. 1967 Aug;99(2):285–290. [PubMed] [Google Scholar]
  18. Shapiro D., Brandt W. E., Cardiff R. D., Russell P. K. The proteins of Japanese encephalitis virus. Virology. 1971 Apr;44(1):108–124. doi: 10.1016/0042-6822(71)90158-9. [DOI] [PubMed] [Google Scholar]
  19. Shapiro D., Brandt W. E., Russell P. K. Change involving a viral membrane glycoprotein during morphogenesis of group B arboviruses. Virology. 1972 Dec;50(3):906–911. doi: 10.1016/0042-6822(72)90445-x. [DOI] [PubMed] [Google Scholar]
  20. Shapiro D., Kos K., Brandt W. E., Russell P. K. Membrane-bound proteins of Japanese encephalitis virus-infected chick embryo cells. Virology. 1972 May;48(2):360–372. doi: 10.1016/0042-6822(72)90046-3. [DOI] [PubMed] [Google Scholar]
  21. Smith T. J., Brandt W. E., Swanson J. L., McCown J. M., Buescher E. L. Physical and biological properties of dengue-2 virus and associated antigens. J Virol. 1970 Apr;5(4):524–532. doi: 10.1128/jvi.5.4.524-532.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Stollar V. Studies on the nature of dengue viruses. IV. The structural proteins of type 2 dengue virus. Virology. 1969 Nov;39(3):426–438. doi: 10.1016/0042-6822(69)90091-9. [DOI] [PubMed] [Google Scholar]

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

RESOURCES