1887

Abstract

Severe acute respiratory syndrome coronavirus (SARS-CoV) contains a single spike (S) protein, which binds to its receptor, angiotensin-converting enzyme 2 (ACE2), induces membrane fusion and serves as a neutralizing antigen. A SARS-CoV-S protein-bearing vesicular stomatitis virus (VSV) pseudotype using the VSVΔG* system was generated. Partial deletion of the SARS-CoV-S protein cytoplasmic domain allowed efficient incorporation into VSV particles and led to the generation of a pseudotype (VSV-SARS-St19) at high titre. Green fluorescent protein expression was demonstrated as early as 7 h after infection of Vero E6 cells with VSV-SARS-St19. VSV-SARS-St19 was neutralized by anti-SARS-CoV antibody and soluble ACE2, and its infection was blocked by treatment of Vero E6 cells with anti-ACE2 antibody. These results indicated that VSV-SARS-St19 infection is mediated by SARS-CoV-S protein in an ACE2-dependent manner. VSV-SARS-St19 will be useful for analysing the function of SARS-CoV-S protein and for developing rapid methods of detecting neutralizing antibodies specific for SARS-CoV infection.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.80955-0
2005-08-01
2019-11-12
Loading full text...

Full text loading...

/deliver/fulltext/jgv/86/8/vir862269.html?itemId=/content/journal/jgv/10.1099/vir.0.80955-0&mimeType=html&fmt=ahah

References

  1. Drosten, C., Gunther, S., Preiser, W. & 23 other authors ( 2003; ). Identification of a novel coronavirus in patients with severe acute respiratory syndrome. N Engl J Med 348, 1967–1976.[CrossRef]
    [Google Scholar]
  2. Fouchier, R. A., Kuiken, T., Schutten, M. & 7 other authors ( 2003; ). Aetiology: Koch's postulates fulfilled for SARS virus. Nature 423, 240.[CrossRef]
    [Google Scholar]
  3. Giroglou, T., Cinatl, J., Jr, Rabenau, H., Drosten, C., Schwalbe, H., Doerr, H. W. & von Laer, D. ( 2004; ). Retroviral vectors pseudotyped with severe acute respiratory syndrome coronavirus S protein. J Virol 78, 9007–9015.[CrossRef]
    [Google Scholar]
  4. Hofmann, H., Geier, M., Marzi, A., Krumbiegel, M., Peipp, M., Fey, G. H., Gramberg, T. & Pohlmann, S. ( 2004; ). Susceptibility to SARS coronavirus S protein-driven infection correlates with expression of angiotensin converting enzyme 2 and infection can be blocked by soluble receptor. Biochem Biophys Res Commun 319, 1216–1221.[CrossRef]
    [Google Scholar]
  5. Huang, L., Sexton, D. J., Skogerson, K. & 13 other authors ( 2003; ). Novel peptide inhibitors of angiotensin-converting enzyme 2. J Biol Chem 278, 15532–15540.[CrossRef]
    [Google Scholar]
  6. Komatsu, T., Suzuki, Y., Imai, J., Sugano, S., Hida, M., Tanigami, A., Muroi, S., Yamada, Y. & Hanaoka, K. ( 2002; ). Molecular cloning, mRNA expression and chromosomal localization of mouse angiotensin-converting enzyme-related carboxypeptidase (mACE2). DNA Seq 13, 217–220.
    [Google Scholar]
  7. Ksiazek, T. G., Erdman, D., Goldsmith, C. S. & 23 other authors ( 2003; ). A novel coronavirus associated with severe acute respiratory syndrome. N Engl J Med 348, 1953–1966.[CrossRef]
    [Google Scholar]
  8. Li, W., Moore, M. J., Vasilieva, N. & 9 other authors ( 2003; ). Angiotensin-converting enzyme 2 is a functional receptor for the SARS coronavirus. Nature 426, 450–454.[CrossRef]
    [Google Scholar]
  9. Marra, M. A., Jones, S. J., Astell, C. R. & 56 other authors ( 2003; ). The genome sequence of the SARS-associated coronavirus. Science 300, 1399–1404.[CrossRef]
    [Google Scholar]
  10. Matsuura, Y., Tani, H., Suzuki, K. & 8 other authors ( 2001; ). Characterization of pseudotype VSV possessing HCV envelope proteins. Virology 286, 263–275.[CrossRef]
    [Google Scholar]
  11. Miura, H. S., Nakagaki, K. & Taguchi, F. ( 2004; ). N-terminal domain of the murine coronavirus receptor CEACAM1 is responsible for fusogenic activation and conformational changes of the spike protein. J Virol 78, 216–223.[CrossRef]
    [Google Scholar]
  12. Mizutani, T., Fukushi, S., Saijo, M., Kurane, I. & Morikawa, S. ( 2004; ). Phosphorylation of p38 MAPK and its downstream targets in SARS coronavirus-infected cells. Biochem Biophys Res Commun 319, 1228–1234.[CrossRef]
    [Google Scholar]
  13. Moore, M. J., Dorfman, T., Li, W. & 9 other authors ( 2004; ). Retroviruses pseudotyped with the severe acute respiratory syndrome coronavirus spike protein efficiently infect cells expressing angiotensin-converting enzyme 2. J Virol 78, 10628–10635.[CrossRef]
    [Google Scholar]
  14. Nagata, S., Okamoto, Y., Inoue, T., Ueno, Y., Kurata, T. & Chiba, J. ( 1992; ). Identification of epitopes associated with different biological activities on the glycoprotein of vesicular stomatitis virus by use of monoclonal antibodies. Arch Virol 127, 153–168.[CrossRef]
    [Google Scholar]
  15. Nie, Y., Wang, P., Shi, X. & 13 other authors ( 2004; ). Highly infectious SARS-CoV pseudotyped virus reveals the cell tropism and its correlation with receptor expression. Biochem Biophys Res Commun 321, 994–1000.[CrossRef]
    [Google Scholar]
  16. Ogino, M., Ebihara, H., Lee, B. H., Araki, K., Lundkvist, A., Kawaoka, Y., Yoshimatsu, K. & Arikawa, J. ( 2003; ). Use of vesicular stomatitis virus pseudotypes bearing Hantaan or Seoul virus envelope proteins in a rapid and safe neutralization test. Clin Diagn Lab Immunol 10, 154–160.
    [Google Scholar]
  17. Rota, P. A., Oberste, M. S., Monroe, S. S. & 32 other authors ( 2003; ). Characterization of a novel coronavirus associated with severe acute respiratory syndrome. Science 300, 1394–1399.[CrossRef]
    [Google Scholar]
  18. Saijo, M., Qing, T., Niikura, M., Maeda, A., Ikegami, T., Sakai, K., Prehaud, C., Kurane, I. & Morikawa, S. ( 2002; ). Immunofluorescence technique using HeLa cells expressing recombinant nucleoprotein for detection of immunoglobulin G antibodies to Crimean-Congo hemorrhagic fever virus. J Clin Microbiol 40, 372–375.[CrossRef]
    [Google Scholar]
  19. Simmons, G., Reeves, J. D., Rennekamp, A. J., Amberg, S. M., Piefer, A. J. & Bates, P. ( 2004; ). Characterization of severe acute respiratory syndrome-associated coronavirus (SARS-CoV) spike glycoprotein-mediated viral entry. Proc Natl Acad Sci U S A 101, 4240–4245.[CrossRef]
    [Google Scholar]
  20. Takada, A., Robison, C., Goto, H., Sanchez, A., Murti, K. G., Whitt, M. A. & Kawaoka, Y. ( 1997; ). A system for functional analysis of Ebola virus glycoprotein. Proc Natl Acad Sci U S A 94, 14764–14769.[CrossRef]
    [Google Scholar]
  21. Tatsuo, H., Okuma, K., Tanaka, K., Ono, N., Minagawa, H., Takade, A., Matsuura, Y. & Yanagi, Y. ( 2000; ). Virus entry is a major determinant of cell tropism of Edmonston and wild-type strains of measles virus as revealed by vesicular stomatitis virus pseudotypes bearing their envelope proteins. J Virol 74, 4139–4145.[CrossRef]
    [Google Scholar]
  22. Zelus, B. D., Wessner, D. R., Williams, R. K., Pensiero, M. N., Phibbs, F. T., deSouza, M., Dveksler, G. S. & Holmes, K. V. ( 1998; ). Purified, soluble recombinant mouse hepatitis virus receptor, Bgp1b, and Bgp2 murine coronavirus receptors differ in mouse hepatitis virus binding and neutralizing activities. J Virol 72, 7237–7244.
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/vir.0.80955-0
Loading
/content/journal/jgv/10.1099/vir.0.80955-0
Loading

Data & Media loading...

Supplements

vol. , part 8, pp. 2269 - 2274

Expression of SARS-CoV-S protein. Expression in insect cells of soluble ACE2 with mutations (soACE2-NN) within the putative catalytic domain. [Single PDF file](138 KB)



PDF

Most Cited This Month

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error