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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.


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