1887

Abstract

A full-length DNA clone of a virulent strain of rinderpest virus was constructed with the gene for the enhanced green fluorescent protein (eGFP) inserted as a separate transcription unit between the P and M genes. Rescue of the virus from the modified clone using reverse genetics generated a virus that grew to the same levels as the virus rescued from the unmodified DNA clone in cell culture. The recombinant virus expressed eGFP to a high level and was used to follow virus replication in real-time using live-cell imaging. Cattle infected with both the recombinant wild-type virus and the recombinant eGFP expressing virus developed clinical disease similar to that of the wild-type natural virus isolate. Detection of virus in circulating peripheral blood leukocytes was equivalent to that of the animals infected with the wild-type virus. The high level of expression of soluble eGFP by this virus allowed us to detect viral replication in infected animals by confocal microscopy. Imaging vibrating microtome sections by confocal microscopy provided good preservation of tissue and cellular architecture as well as revealing the sites of replication of the virus in different tissues of infected animals.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.023598-0
2010-12-01
2019-11-14
Loading full text...

Full text loading...

/deliver/fulltext/jgv/91/12/2918.html?itemId=/content/journal/jgv/10.1099/vir.0.023598-0&mimeType=html&fmt=ahah

References

  1. Anderson, J., Barrett, T. & Scott, G. R. ( 1996; ). ). FAO Animal Health Manual -Manual on the Diagnosis of Rinderpest, 2nd edn, pp. 81–91. Rome. : FAO.
    [Google Scholar]
  2. Banyard, A. C., Baron, M. D. & Barrett, T. ( 2005; ). A role for virus promoters in determining the pathogenesis of Rinderpest virus in cattle. J Gen Virol 86, 1083–1092.[CrossRef]
    [Google Scholar]
  3. Banyard, A., Rima, B. & Barrett, T. ( 2006; ). The morbilliviruses. In Rinderpest and peste des petits ruminants virus, pp. 13–26. Edited by Barrett, T., Pastouret, P. P. & Taylor, W. P.. London. : Academic Press, Elsevier.
    [Google Scholar]
  4. Baron, M. D. & Barrett, T. ( 1997; ). Rescue of rinderpest virus from cloned cDNA. J Virol 71, 1265–1271.
    [Google Scholar]
  5. Baron, M. D., Foster-Cuevas, M., Baron, J. & Barrett, T. ( 1999; ). Expression in cattle of epitopes of a heterologous virus using a recombinant rinderpest virus. J Gen Virol 80, 2031–2039.
    [Google Scholar]
  6. Baron, M. D., Banyard, A. C., Parida, S. & Barrett, T. ( 2005; ). The Plowright vaccine strain of rinderpest virus has attenuating mutations in most genes. J Gen Virol 86, 1093–1101.[CrossRef]
    [Google Scholar]
  7. Barrett, T., Banyard, A. C. & Diallo, A. ( 2006; ). Molecular biology of the morbilliviruses. In Rinderpest and peste des petits ruminants virus, pp. 31–56. Edited by Barrett, T., Pastouret, P. P. & Taylor, W. P.. London. : Academic Press, Elsevier.
    [Google Scholar]
  8. Britton, P., Green, P., Kottier, S., Mawditt, K. L., Penzes, P., Cavanagh, D. & Skinner, M. A. ( 1996; ). Expression of bacteriophage T7 RNA polymerase in avian and mammalian cells by a recombinant fowlpox virus. J Gen Virol 77, 963–967.[CrossRef]
    [Google Scholar]
  9. Brown, D. D., Rima, B. K., Allen, I. V., Baron, M. D., Banyard, A. C., Barrett, T. & Duprex, W. P. ( 2005; ). Rational attenuation of a morbillivirus by modulating the activity of the RNA-dependent RNA polymerase. J Virol 79, 14330–14338.[CrossRef]
    [Google Scholar]
  10. Cosby, S., Kai, C. & Yamanouchi, K. ( 2006; ). Immunology of Rinderpest – an immunosuppression but a lifelong vaccine protection. In Rinderpest and peste des petits ruminants virus, pp. 196–216. Edited by Barrett, T., Pastouret, P. P. & Taylor, W. P.. London. : Academic Press, Elsevier.
    [Google Scholar]
  11. Das, S. C., Baron, M. D., Skinner, M. A. & Barrett, T. ( 2000; ). Improved technique for transient expression and negative strand virus rescue using fowlpox T7 recombinant virus in mammalian cells. J Virol Methods 89, 119–127.[CrossRef]
    [Google Scholar]
  12. Das, S. C., Nayak, D., Zhou, Y. & Pattnaik, A. K. ( 2006; ). Visualization of intracellular transport of vesicular stomatitis virus nucleocapsids in living cells. J Virol 80, 6368–6377.[CrossRef]
    [Google Scholar]
  13. de Vries, R. D., Lemon, K., Ludlow, M., McQuaid, S., Yuksel, S., van Amerongen, G., Rennick, L. J., Rima, B. K., Osterhaus, A. D. & other authors ( 2010; ). In vivo tropism of attenuated and pathogenic measles virus expressing green fluorescent protein in macaques. J Virol 84, 4714–4724.[CrossRef]
    [Google Scholar]
  14. Duprex, W. P., McQuaid, S., Hangartner, L., Billiter, M. A. & Rima, B. K. ( 1999; ). Observation of measles virus cell-to-cell spread in astrcytoma cells by using a green fluorescent protein-expressing recombinant virus. J Virol 73, 9568–9575.
    [Google Scholar]
  15. Duprex, W. P., McQuaid, S., Roscic-Mrkic, B., Cattaneo, R., McCallister, C. & Rima, B. K. ( 2000; ). In vitro and in vivo infection of neural cells by a recombinant measles virus expressing enhanced green fluorescent protein. J Virol 74, 7972–7979.[CrossRef]
    [Google Scholar]
  16. Duprex, W. P., Collins, F. M. & Rima, B. K. ( 2002; ). Modulating the function of the measles virus RNA-dependent RNA polymerase by insertion of green fluorescent protein into the open reading frame. J Virol 76, 7322–7328.[CrossRef]
    [Google Scholar]
  17. Forsyth, M. A. & Barrett, T. ( 1995; ). Evaluation of polymerase chain reaction for the detection and characterisation of rinderpest and peste des petits ruminants viruses for epidemiological studies. Virus Res 39, 151–163.[CrossRef]
    [Google Scholar]
  18. Fuerst, T. R., Niles, E. G., Studier, F. W. & Moss, B. ( 1986; ). Eukaryotic transient-expression system based on recombinant vaccinia virus that synthesizes bacteriophage-T7 RNA-polymerase. Proc Natl Acad Sci U S A 83, 8122–8126.[CrossRef]
    [Google Scholar]
  19. Huang, Z., Elankumaran, S., Yunus, A. S. & Samal, S. K. ( 2004; ). A recombinant Newcastle disease virus (NDV) expressing VP2 protein of infectious bursal disease virus (IBDV) protects against NDV and IBDV. J Virol 78, 10054–10063.[CrossRef]
    [Google Scholar]
  20. Kobune, F., Sakata, H., Sugiyama, M. & Sugiura, A. ( 1991; ). B95a, a marmoset lymphoblastoid cell line, as a sensitive host for rinderpest virus. J Gen Virol 72, 687–692.[CrossRef]
    [Google Scholar]
  21. Ohishi, K., Inui, K., Yamanouchi, K. & Barrett, T. ( 1999; ). Cell-mediated immune responses in cattle vaccinated with a vaccinia recombinant expressing the nucleocapsid (N) protein of rinderpest virus. J Gen Virol 80, 1627–1634.
    [Google Scholar]
  22. Panda, A., Huang, Z., Elankumaran, S., Rockemann, D. D. & Samal, S. K. ( 2004; ). Role of fusion protein cleavage site in the virulence of Newcastle disease virus. Microb Pathog 36, 1–10.[CrossRef]
    [Google Scholar]
  23. Parida, S., Mahapatra, M., Kumar, S., Das, S. C., Baron, M. D., Anderson, J. & Barrett, T. ( 2007; ). Rescue of a chimeric rinderpest virus with the nucleocapsid protein derived from peste-des-petits-ruminants virus: use as a marker vaccine. J Gen Virol 88, 2019–2027.[CrossRef]
    [Google Scholar]
  24. Plattet, P., Zweifel, C., Wiederkehr, C., Belloy, L., Cherpillod, P., Zurbriggen, A. & Wittek, R. ( 2004; ). Recovery of a persistent Canine distemper virus expressing the enhanced green fluorescent protein from cloned cDNA. Virus Res 101, 147–153.[CrossRef]
    [Google Scholar]
  25. Reed, L. J. & Muench, H. ( 1938; ). A simple method of estimating fifty per cent endpoints. Am J Hyg 27, 493–497.
    [Google Scholar]
  26. Silin, D., Lyubomska, O., Ludlow, M., Duprex, W. P. & Rima, B. K. ( 2007; ). Development of a challenge-protective vaccine concept by modification of the viral RNA-dependent RNA polymerase of canine distemper virus. J Virol 81, 13649–13658.[CrossRef]
    [Google Scholar]
  27. Sinnathamby, G., Seth, S., Nayak, R. & Shaila, M. S. ( 2004; ). Cytotoxic T cell epitope in cattle from the attachment glycoproteins of rinderpest and peste des petits ruminants viruses. Viral Immuno 17, 401–410.[CrossRef]
    [Google Scholar]
  28. Taylor, W. P. ( 1986; ). Epidemiology and control of rinderpest. Rev Off Int Epizoot 5, 407–410.
    [Google Scholar]
  29. Taylor, W. P., Plowright, W., Pillinger, R., Rampton, C. S. & Staple, R. F. ( 1965; ). Studies on the pathogenesis of rinderpest in experimental cattle. IV. Proliferation of the virus following contact infection. J Hyg (Lond) 63, 497–506.[CrossRef]
    [Google Scholar]
  30. von Messling, V., Springfeld, C., Devaux, P. & Cattaneo, R. ( 2003; ). A ferret model of canine distemper virus virulence and immunosuppression. J Virol 77, 12579–12591.[CrossRef]
    [Google Scholar]
  31. Walsh, E. P., Baron, M. D., Anderson, J. & Barrett, T. ( 2000a; ). Development of a genetically marked recombinant rinderpest vaccine expressing green fluorescent protein. J Gen Virol 81, 709–718.
    [Google Scholar]
  32. Walsh, E. P., Baron, M. D., Rennie, L. F., Monaghan, P., Anderson, J. & Barrett, T. ( 2000b; ). Recombinant rinderpest vaccines expressing membrane-anchored proteins as genetic markers: evidence of exclusion of marker protein from the virus envelope. J Virol 74, 10165–10175.[CrossRef]
    [Google Scholar]
  33. Wamwayi, H. M., Fleming, M. & Barrett, T. ( 1995; ). Characterisation of African isolates of rinderpest virus. Vet Microbiol 44, 151–163.[CrossRef]
    [Google Scholar]
  34. Wertz, G. W., Moudy, R. & Ball, L. A. ( 2002; ). Adding genes to the RNA genome of VSV: positional effects on stability of expression. J Virol 76, 7642–7650.[CrossRef]
    [Google Scholar]
  35. Wertz, G. W., Perepelitsa, V. P. & Ball, L. A. ( 1998; ). Gene rearrangement attenuates expression and lethality of a nonsegmented negative strand RNA virus. Proc Natl Acad Sci U S A 95, 3501–3506.[CrossRef]
    [Google Scholar]
  36. Wohlsein, P., Wamwayi, H. M., Trautwein, G., Pohlenz, J., Liess, B. & Barrett, T. ( 1995; ). Pathomorphological and immunohistological findings in cattle experimentally infected with rinderpest virus isolates of different pathogenicity. Vet Microbiol 44, 141–149.[CrossRef]
    [Google Scholar]
  37. Yoneda, M., Bandyopadhyay, S. K., Shiotani, M., Fujita, K., Nuntaprasert, A., Miura, R., Baron, M. D., Barrett, T. & Kai, C. ( 2002; ). Rinderpest virus H protein: role in determining host range in rabbits. J Gen Virol 83, 1457–1463.
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/vir.0.023598-0
Loading
/content/journal/jgv/10.1099/vir.0.023598-0
Loading

Data & Media loading...

Supplements

rKOIns–eGFP B95a infection of B95a cells; 27–42 h p.i.; 512x512 resolution, magnification x40 [avi](36 MB)

[avi] 

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