This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Boot, H. J.
Right arrow Articles by Gielkens, A. L. J.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Boot, H. J.
Right arrow Articles by Gielkens, A. L. J.

Next Article 

Journal of Virology, August 2000, p. 6701-6711, Vol. 74, No. 15
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Rescue of Very Virulent and Mosaic Infectious Bursal Disease Virus from Cloned cDNA: VP2 Is Not the Sole Determinant of the Very Virulent Phenotype

Hein J. Boot,* A. Agnes H. M. ter Huurne, Arjan J. W. Hoekman, Ben P. H. Peeters, and Arno L. J. Gielkens

Department of Avian Virology, Institute for Animal Science and Health, Lelystad, The Netherlands

Received 17 November 1999/Accepted 18 March 2000

Many recent outbreaks of infectious bursal disease in commercial chicken flocks worldwide are due to the spread of very virulent strains of infectious bursal disease virus (vvIBDV). The molecular determinants for the enhanced virulence of vvIBDV compared to classical IBDV are unknown. The lack of a reverse genetics system to rescue vvIBDV from its cloned cDNA hampers the identification and study of these determinants. In this report we describe, for the first time, the rescue of vvIBDV from its cloned cDNA. Two plasmids containing a T7 promoter and either the full-length A- or B-segment cDNA of vvIBDV (D6948) were cotransfected into QM5 cells expressing T7 polymerase. The presence of vvIBDV could be detected after passage of the transfection supernatant in either primary bursa cells (in vitro) or embryonated eggs (in vivo), but not QM5 cells. Rescued vvIBDV (rD6948) appeared to have the same virulence as the parental isolate, D6948. Segment-reassorted IBDV, in which one of the two genomic segments originated from cDNA of classical attenuated IBDV CEF94 and the other from D6948, could also be rescued by using this system. Segment-reassorted virus containing the A segment of the classical attenuated isolate (CEF94) and the B segment of the very virulent isolate (D6948) is not released until 15 h after an in vitro infection. This indicates a slightly retarded replication, as the first release of CEF94 is already found at 10 h after infection. Next to segment reassortants, we generated and analyzed mosaic IBDVs (mIBDVs). In these mIBDVs we replaced the region of CEF94 encoding one of the viral proteins (pVP2, VP3, or VP4) by the corresponding region of D6948. Analysis of these mIBDV isolates showed that tropism for non-B-lymphoid cells was exclusively determined by the viral capsid protein VP2. However, the very virulent phenotype was not solely determined by this protein, since mosaic virus containing VP2 of vvIBDV induced neither morbidity nor mortality in young chickens.

* Corresponding author. Mailing address: ID-Lelystad, P.O. Box 65, NL-8200 AB Lelystad, The Netherlands. Phone: 31 320 238 695. Fax: 31 320 238 668. E-mail: H.J.Boot{at}id.wag-ur.nl.

Journal of Virology, August 2000, p. 6701-6711, Vol. 74, No. 15
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Hon, C.-C., Lam, T.-Y., Drummond, A., Rambaut, A., Lee, Y.-F., Yip, C.-W., Zeng, F., Lam, P.-Y., Ng, P. T. W., Leung, F. C. C. (2006). Phylogenetic Analysis Reveals a Correlation between the Expansion of Very Virulent Infectious Bursal Disease Virus and Reassortment of Its Genome Segment B.. J. Virol. 80: 8503-8509 [Abstract] [Full Text]  
  • Nouen, C. L., Rivallan, G., Toquin, D., Darlu, P., Morin, Y., Beven, V., de Boisseson, C., Cazaban, C., Comte, S., Gardin, Y., Eterradossi, N. (2006). Very virulent infectious bursal disease virus: reduced pathogenicity in a rare natural segment-B-reassorted isolate. J. Gen. Virol. 87: 209-216 [Abstract] [Full Text]  
  • Boot, H. J., Pritz-Verschuren, S. B. E. (2004). Modifications of the 3'-UTR stem-loop of infectious bursal disease virus are allowed without influencing replication or virulence. Nucleic Acids Res 32: 211-222 [Abstract] [Full Text]  
  • Dieye, Y., Hoekman, A. J. W., Clier, F., Juillard, V., Boot, H. J., Piard, J.-C. (2003). Ability of Lactococcus lactis To Export Viral Capsid Antigens: a Crucial Step for Development of Live Vaccines. Appl. Environ. Microbiol. 69: 7281-7288 [Abstract] [Full Text]  
  • Da Costa, B., Soignier, S., Chevalier, C., Henry, C., Thory, C., Huet, J.-C., Delmas, B. (2002). Blotched Snakehead Virus Is a New Aquatic Birnavirus That Is Slightly More Related to Avibirnavirus Than to Aquabirnavirus. J. Virol. 77: 719-725 [Abstract] [Full Text]  
  • Tacken, M. G. J., Peeters, B. P. H., Thomas, A. A. M., Rottier, P. J. M., Boot, H. J. (2002). Infectious Bursal Disease Virus Capsid Protein VP3 Interacts both with VP1, the RNA-Dependent RNA Polymerase, and with Viral Double-Stranded RNA. J. Virol. 76: 11301-11311 [Abstract] [Full Text]  
  • Boot, H. J., ter Huurne, A. A. H. M., Hoekman, A. J. W., Pol, J. M., Gielkens, A. L. J., Peeters, B. P. H. (2002). Exchange of the C-Terminal Part of VP3 from Very Virulent Infectious Bursal Disease Virus Results in an Attenuated Virus with a Unique Antigenic Structure. J. Virol. 76: 10346-10355 [Abstract] [Full Text]  
  • van Loon, A. A. W. M., de Haas, N., Zeyda, I., Mundt, E. (2002). Alteration of amino acids in VP2 of very virulent infectious bursal disease virus results in tissue culture adaptation and attenuation in chickens. J. Gen. Virol. 83: 121-129 [Abstract] [Full Text]  
  • Brandt, M., Yao, K., Liu, M., Heckert, R. A., Vakharia, V. N. (2001). Molecular Determinants of Virulence, Cell Tropism, and Pathogenic Phenotype of Infectious Bursal Disease Virus. J. Virol. 75: 11974-11982 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»