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 Widjojoatmodjo, M. N.
Right arrow Articles by Moormann, R. J. M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Widjojoatmodjo, M. N.
Right arrow Articles by Moormann, R. J. M.

 Previous Article  |  Next Article 

Journal of Virology, April 2000, p. 2973-2980, Vol. 74, No. 7
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Classical Swine Fever Virus Erns Deletion Mutants: trans-Complementation and Potential Use as Nontransmissible, Modified, Live-Attenuated Marker Vaccines

M. N. Widjojoatmodjo,dagger H. G. P. van Gennip, A. Bouma, P. A. van Rijn, and R. J. M. Moormann*

Department of Mammalian Virology, DLO-Institute for Animal Science and Health, Lelystad, The Netherlands

Received 30 August 1999/Accepted 21 December 1999

An SK6 cell line (SK6c26) which constitutively expressed the glycoprotein Erns of classical swine fever virus (CSFV) was used to rescue CSFV Erns deletion mutants based on the infectious copy of CSFV strain C. The biochemical properties of Erns from this cell line were indistinguishable from those of CSFV Erns. Two Erns deletion mutants were constructed, virus Flc23 and virus Flc22. Virus Flc23 encoded only the utmost N- and C-terminal amino acids of Erns (deletion of 215 amino acids) to retain the original protease cleavage sites. Virus Flc22 is not recognized by a panel of Erns antibodies, due to a deletion of 66 amino acids in Erns. The Erns deletion mutants Flc22 and Flc23 could be rescued in vitro only on the complementing SK6c26 cells. These rescued viruses could infect and replicate in SK6 cells but did not yield infectious virus. Virus neutralization by Erns-specific antibodies was similar for the wild-type virus and the recombinant viruses, indicating that Erns from SK6c26 cells was incorporated in the viral particles. Pigs vaccinated with Flc22 or Flc23 were protected against a challenge with a lethal dose of CSFV strain Brescia. This is the first demonstration of trans-complementation of defective pestivirus RNA with a pestiviral structural protein and opens new ways to develop nontransmissible modified live pestivirus vaccines. In addition, the absence of (the antigenic part of) Erns in the recombinant viral particles can be used to differentiate between infected and vaccinated animals.


* Corresponding author. Mailing address: Department of Mammalian Virology, DLO-Institute for Animal Science and Health, P.O. Box 65, Lelystad, NL-8200 AB, The Netherlands. Phone: 31-320-238238. Fax: 31-320-238668. E-mail: r.j.m.moormann{at}id.wag-ur.nl.

dagger Present address: RIVM, National Institute for Public Health and the Environment, Bilthoven, The Netherlands.


Journal of Virology, April 2000, p. 2973-2980, Vol. 74, No. 7
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.



This article has been cited by other articles:

  • Wegelt, A., Reimann, I., Zemke, J., Beer, M. (2009). New insights into processing of bovine viral diarrhea virus glycoproteins Erns and E1. J. Gen. Virol. 90: 2462-2467 [Abstract] [Full Text]  
  • Tews, B. A., Schurmann, E.-M., Meyers, G. (2009). Mutation of Cysteine 171 of Pestivirus Erns RNase Prevents Homodimer Formation and Leads to Attenuation of Classical Swine Fever Virus. J. Virol. 83: 4823-4834 [Abstract] [Full Text]  
  • Fang, Y., Christopher-Hennings, J., Brown, E., Liu, H., Chen, Z., Lawson, S. R., Breen, R., Clement, T., Gao, X., Bao, J., Knudsen, D., Daly, R., Nelson, E. (2008). Development of genetic markers in the non-structural protein 2 region of a US type 1 porcine reproductive and respiratory syndrome virus: implications for future recombinant marker vaccine development. J. Gen. Virol. 89: 3086-3096 [Abstract] [Full Text]  
  • Wehrle, F., Renzullo, S., Faust, A., Beer, M., Kaden, V., Hofmann, M. A. (2007). Chimeric pestiviruses: candidates for live-attenuated classical swine fever marker vaccines. J. Gen. Virol. 88: 2247-2258 [Abstract] [Full Text]  
  • Meyers, G., Ege, A., Fetzer, C., von Freyburg, M., Elbers, K., Carr, V., Prentice, H., Charleston, B., Schurmann, E.-M. (2007). Bovine Viral Diarrhea Virus: Prevention of Persistent Fetal Infection by a Combination of Two Mutations Affecting Erns RNase and Npro Protease. J. Virol. 81: 3327-3338 [Abstract] [Full Text]  
  • Ceppi, M., de Bruin, M. G. M., Seuberlich, T., Balmelli, C., Pascolo, S., Ruggli, N., Wienhold, D., Tratschin, J. D., McCullough, K. C., Summerfield, A. (2005). Identification of classical swine fever virus protein E2 as a target for cytotoxic T cells by using mRNA-transfected antigen-presenting cells. J. Gen. Virol. 86: 2525-2534 [Abstract] [Full Text]  
  • Risatti, G. R., Borca, M. V., Kutish, G. F., Lu, Z., Holinka, L. G., French, R. A., Tulman, E. R., Rock, D. L. (2005). The E2 Glycoprotein of Classical Swine Fever Virus Is a Virulence Determinant in Swine. J. Virol. 79: 3787-3796 [Abstract] [Full Text]  
  • van Gennip, H. G. P., Vlot, A. C., Hulst, M. M., de Smit, A. J., Moormann, R. J. M. (2004). Determinants of Virulence of Classical Swine Fever Virus Strain Brescia. J. Virol. 78: 8812-8823 [Abstract] [Full Text]  
  • von Freyburg, M., Ege, A., Saalmuller, A., Meyers, G. (2004). Comparison of the effects of RNase-negative and wild-type classical swine fever virus on peripheral blood cells of infected pigs. J. Gen. Virol. 85: 1899-1908 [Abstract] [Full Text]  
  • Hausmann, Y., Roman-Sosa, G., Thiel, H.-J., Rumenapf, T. (2004). Classical Swine Fever Virus Glycoprotein Erns Is an Endoribonuclease with an Unusual Base Specificity. J. Virol. 78: 5507-5512 [Abstract] [Full Text]  
  • Castillo-Olivares, J., Wieringa, R., Bakonyi, T., de Vries, A. A. F., Davis-Poynter, N. J., Rottier, P. J. M. (2003). Generation of a Candidate Live Marker Vaccine for Equine Arteritis Virus by Deletion of the Major Virus Neutralization Domain. J. Virol. 77: 8470-8480 [Abstract] [Full Text]  
  • Mebatsion, T., Koolen, M. J. M., de Vaan, L. T. C., de Haas, N., Braber, M., Romer-Oberdorfer, A., van den Elzen, P., van der Marel, P. (2002). Newcastle Disease Virus (NDV) Marker Vaccine: an Immunodominant Epitope on the Nucleoprotein Gene of NDV Can Be Deleted or Replaced by a Foreign Epitope. J. Virol. 76: 10138-10146 [Abstract] [Full Text]