Skip to main content
NCBI home page
Journal of Virology logoLink to Journal of Virology
. 1995 Apr;69(4):2667–2673. doi: 10.1128/jvi.69.4.2667-2673.1995

Respiratory syncytial virus matures at the apical surfaces of polarized epithelial cells.

S R Roberts 1, R W Compans 1, G W Wertz 1
PMCID: PMC188952  PMID: 7884920

Abstract

Respiratory syncytial (RS) virus infects the epithelium of the respiratory tract. We examined the replication and maturation of RS virus in two polarized epithelial cell lines, Vero C1008 and MDCK. Electron microscopy of RS virus-infected Vero C1008 cells revealed the presence of pleomorphic viral particles budding exclusively from the apical surface, often in clusters. The predominant type of particle was filamentous, 80 to 100 nm in diameter, and 4 to 8 microns in length, and evidence from filtration studies indicated that the filamentous particles were infectious. Cytopathology produced by RS virus infection of polarized Vero C1008 cells was minimal, and syncytia were not observed, consistent with the maintenance of tight junctions and the exclusively apical maturation of the virus. Infectivity assays with MDCK cells confirmed that in this cell line, RS virus was released into the apical medium but not into the basolateral medium. In addition, the majority of the RS virus transmembrane fusion glycoprotein on the cell surface was localized to the apical surface of the Vero C1008 cells. Taken together, these results demonstrate that RS virus matures at the apical surface of polarized epithelial cell lines.

Full Text

The Full Text of this article is available as a PDF (2.3 MB).

Selected References

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

  1. Aroeti B., Henis Y. I. Accumulation of Sendai virus glycoproteins in cell-cell contact regions and its role in cell fusion. J Biol Chem. 1991 Aug 25;266(24):15845–15849. [PubMed] [Google Scholar]
  2. Berthiaume L., Joncas J., Pavilanis V. Comparative structure, morphogenesis and biological characteristics of the respiratory syncytial (RS) virus and the pneumonia virus of mice (PVM). Arch Gesamte Virusforsch. 1974;45(1-2):39–51. doi: 10.1007/BF01240540. [DOI] [PubMed] [Google Scholar]
  3. Bryson D. G., McConnell S., McAliskey M., McNulty M. S. Ultrastructural features of alveolar lesions in induced respiratory syncytial virus pneumonia of calves. Vet Pathol. 1991 Jul;28(4):286–292. doi: 10.1177/030098589102800404. [DOI] [PubMed] [Google Scholar]
  4. Bryson D. G., Platten M. F., McConnell S., McNulty M. S. Ultrastructural features of lesions in bronchiolar epithelium in induced respiratory syncytial virus pneumonia of calves. Vet Pathol. 1991 Jul;28(4):293–299. doi: 10.1177/030098589102800405. [DOI] [PubMed] [Google Scholar]
  5. Bächi T. Direct observation of the budding and fusion of an enveloped virus by video microscopy of viable cells. J Cell Biol. 1988 Nov;107(5):1689–1695. doi: 10.1083/jcb.107.5.1689. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bächi T., Howe C. Morphogenesis and ultrastructure of respiratory syncytial virus. J Virol. 1973 Nov;12(5):1173–1180. doi: 10.1128/jvi.12.5.1173-1180.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chen S. Y., Matsuoka Y., Compans R. W. Assembly and polarized release of Punta Toro virus and effects of brefeldin A. J Virol. 1991 Mar;65(3):1427–1439. doi: 10.1128/jvi.65.3.1427-1439.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Clayson E. T., Brando L. V., Compans R. W. Release of simian virus 40 virions from epithelial cells is polarized and occurs without cell lysis. J Virol. 1989 May;63(5):2278–2288. doi: 10.1128/jvi.63.5.2278-2288.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Compans R. W., Harter D. H., Choppin P. W. Studies on pneumonia virus of mice (PVM) in cell culture. II. Structure and morphogenesis of the virus particle. J Exp Med. 1967 Aug 1;126(2):267–276. doi: 10.1084/jem.126.2.267. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Compans R. W., Srinivas R. V. Protein sorting in polarized epithelial cells. Curr Top Microbiol Immunol. 1991;170:141–181. doi: 10.1007/978-3-642-76389-2_5. [DOI] [PubMed] [Google Scholar]
  11. Faulkner G. P., Shirodaria P. V., Follett E. A., Pringle C. R. Respiratory syncytial virus ts mutants and nuclear immunofluorescence. J Virol. 1976 Nov;20(2):487–500. doi: 10.1128/jvi.20.2.487-500.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fuchs H., Bächi T. Scanning electron microscopical demonstration of respiratory syncytial virus antigens by immunological markers. J Ultrastruct Res. 1975 Jul;52(1):114–119. doi: 10.1016/s0022-5320(75)80026-8. [DOI] [PubMed] [Google Scholar]
  13. Fuller S., von Bonsdorff C. H., Simons K. Vesicular stomatitis virus infects and matures only through the basolateral surface of the polarized epithelial cell line, MDCK. Cell. 1984 Aug;38(1):65–77. doi: 10.1016/0092-8674(84)90527-0. [DOI] [PubMed] [Google Scholar]
  14. Gstraunthaler G. J. Epithelial cells in tissue culture. Ren Physiol Biochem. 1988 Jan-Feb;11(1-2):1–42. doi: 10.1159/000173147. [DOI] [PubMed] [Google Scholar]
  15. Heilman C. A. From the National Institute of Allergy and Infectious Diseases and the World Health Organization. Respiratory syncytial and parainfluenza viruses. J Infect Dis. 1990 Mar;161(3):402–406. doi: 10.1093/infdis/161.3.402. [DOI] [PubMed] [Google Scholar]
  16. Joncas J., Berthiaume L., Pavilanis V. The structure of the respiratory syncytial virus. Virology. 1969 Jul;38(3):493–496. doi: 10.1016/0042-6822(69)90166-4. [DOI] [PubMed] [Google Scholar]
  17. Kalica A. R., Wright P. F., Hetrick F. M., Chanock R. M. Electron microscopic studies of respiratory syncytial temperature-sensitive mutants. Arch Gesamte Virusforsch. 1973;41(3):248–258. doi: 10.1007/BF01252772. [DOI] [PubMed] [Google Scholar]
  18. Norrby E., Marusyk H., Orvell C. Morphogenesis of respiratory syncytial virus in a green monkey kidney cell line (Vero). J Virol. 1970 Aug;6(2):237–242. doi: 10.1128/jvi.6.2.237-242.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Owens R. J., Compans R. W. Expression of the human immunodeficiency virus envelope glycoprotein is restricted to basolateral surfaces of polarized epithelial cells. J Virol. 1989 Feb;63(2):978–982. doi: 10.1128/jvi.63.2.978-982.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Owens R. J., Dubay J. W., Hunter E., Compans R. W. Human immunodeficiency virus envelope protein determines the site of virus release in polarized epithelial cells. Proc Natl Acad Sci U S A. 1991 May 1;88(9):3987–3991. doi: 10.1073/pnas.88.9.3987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Paccaud M. F., Jacquier C. A respiratory syncytial virus of bovine origin. Arch Gesamte Virusforsch. 1970;30(4):327–342. doi: 10.1007/BF01258363. [DOI] [PubMed] [Google Scholar]
  22. Parry J. E., Shirodaria P. V., Pringle C. R. Pneumoviruses: the cell surface of lytically and persistently infected cells. J Gen Virol. 1979 Aug;44(2):479–491. doi: 10.1099/0022-1317-44-2-479. [DOI] [PubMed] [Google Scholar]
  23. Roberts S. R., Lichtenstein D., Ball L. A., Wertz G. W. The membrane-associated and secreted forms of the respiratory syncytial virus attachment glycoprotein G are synthesized from alternative initiation codons. J Virol. 1994 Jul;68(7):4538–4546. doi: 10.1128/jvi.68.7.4538-4546.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Rodriguez Boulan E., Pendergast M. Polarized distribution of viral envelope proteins in the plasma membrane of infected epithelial cells. Cell. 1980 May;20(1):45–54. doi: 10.1016/0092-8674(80)90233-0. [DOI] [PubMed] [Google Scholar]
  25. Rodriguez Boulan E., Sabatini D. D. Asymmetric budding of viruses in epithelial monlayers: a model system for study of epithelial polarity. Proc Natl Acad Sci U S A. 1978 Oct;75(10):5071–5075. doi: 10.1073/pnas.75.10.5071. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Rodriguez-Boulan E., Nelson W. J. Morphogenesis of the polarized epithelial cell phenotype. Science. 1989 Aug 18;245(4919):718–725. doi: 10.1126/science.2672330. [DOI] [PubMed] [Google Scholar]
  27. Roth M. G., Srinivas R. V., Compans R. W. Basolateral maturation of retroviruses in polarized epithelial cells. J Virol. 1983 Mar;45(3):1065–1073. doi: 10.1128/jvi.45.3.1065-1073.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Srinivas R. V., Balachandran N., Alonso-Caplen F. V., Compans R. W. Expression of herpes simplex virus glycoproteins in polarized epithelial cells. J Virol. 1986 May;58(2):689–693. doi: 10.1128/jvi.58.2.689-693.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Tashiro M., Seto J. T., Choosakul S., Yamakawa M., Klenk H. D., Rott R. Budding site of Sendai virus in polarized epithelial cells is one of the determinants for tropism and pathogenicity in mice. Virology. 1992 Apr;187(2):413–422. doi: 10.1016/0042-6822(92)90443-s. [DOI] [PubMed] [Google Scholar]
  30. Tashiro M., Yamakawa M., Tobita K., Klenk H. D., Rott R., Seto J. T. Organ tropism of Sendai virus in mice: proteolytic activation of the fusion glycoprotein in mouse organs and budding site at the bronchial epithelium. J Virol. 1990 Aug;64(8):3627–3634. doi: 10.1128/jvi.64.8.3627-3634.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Tashiro M., Yamakawa M., Tobita K., Klenk H. D., Seto J. T., Rott R. Significance of basolateral domain of polarized MDCK cells for Sendai virus-induced cell fusion. Arch Virol. 1992;125(1-4):129–139. doi: 10.1007/BF01309633. [DOI] [PubMed] [Google Scholar]
  32. Tashiro M., Yamakawa M., Tobita K., Seto J. T., Klenk H. D., Rott R. Altered budding site of a pantropic mutant of Sendai virus, F1-R, in polarized epithelial cells. J Virol. 1990 Oct;64(10):4672–4677. doi: 10.1128/jvi.64.10.4672-4677.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Tucker S. P., Compans R. W. Virus infection of polarized epithelial cells. Adv Virus Res. 1993;42:187–247. doi: 10.1016/S0065-3527(08)60086-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Tucker S. P., Melsen L. R., Compans R. W. Migration of polarized epithelial cells through permeable membrane substrates of defined pore size. Eur J Cell Biol. 1992 Aug;58(2):280–290. [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES