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Journal of Virology, August 2003, p. 8470-8480, Vol. 77, No. 15
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.15.8470-8480.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Generation of a Candidate Live Marker Vaccine for Equine Arteritis Virus by Deletion of the Major Virus Neutralization Domain

Javier Castillo-Olivares,1 Roeland Wieringa,2 Tamás Bakonyi,2,{dagger} Antoine A. F. de Vries,2,{ddagger} Nick J. Davis-Poynter,1 and Peter J. M. Rottier2*

Centre for Preventive Medicine, Animal Health Trust, Kentford, Newmarket, Suffolk CB8 7UU, United Kingdom,1 Virology Division, Department of Infectious Diseases and Immunology, Faculty of Veterinary Medicine, and Institute of Biomembranes, Utrecht University, 3584 CL Utrecht, The Netherlands2

Received 10 January 2003/ Accepted 13 May 2003

Equine arteritis virus (EAV) is an enveloped plus-strand RNA virus of the family Arteriviridae (order Nidovirales) that causes respiratory and reproductive disease in equids. Protective, virus-neutralizing antibodies (VNAb) elicited by infection are directed predominantly against an immunodominant region in the membrane-proximal domain of the viral envelope glycoprotein GL, allowing recently the establishment of a sensitive peptide enzyme-linked immunosorbent assay (ELISA) based on this particular domain (J. Nugent et al., J. Virol. Methods 90:167-183, 2000). By using an infectious cDNA we have now generated, in the controlled background of a nonvirulent virus, a mutant EAV from which this immunodominant domain was deleted. This virus, EAV-GL{Delta}, replicated to normal titers in culture cells, although at a slower rate than wild-type EAV, and caused an asymptomatic infection in ponies. The antibodies induced neutralized the mutant virus efficiently in vitro but reacted poorly to wild-type EAV strains. Nevertheless, when inoculated subsequently with virulent EAV, the immunized animals, in contrast to nonvaccinated controls, were fully protected against disease; replication of the challenge virus occurred briefly at low though detectable levels. The levels of protection achieved suggest that an immune effector mechanism other than VNAb plays an important role in protection against infection. As expected, infection with EAV-GL{Delta} did not induce a measurable response in our GL-peptide ELISA while the challenge infection of the animals clearly did. EAV-GL{Delta} or similar mutants are therefore attractive marker vaccine candidates, enabling serological discrimination between vaccinated and wild-type virus-infected animals.

* Corresponding author. Mailing address: Virology Division, Department of Infectious Diseases and Immunology, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 1, 3584 CL Utrecht, The Netherlands. Phone: 31-30-253 2462. Fax: 31-30-253 6723. E-mail: p.rottier{at}vet.uu.nl.

{dagger} Present address: Department of Microbiology and Infectious Diseases, Faculty of Veterinary Sciences, Szent Istvan University, Budapest, Hungary.

{ddagger} Present address: Gene Therapy Section, Department of Molecular Cell Biology, Leiden University Medical Center, 2333 AL Leiden, The Netherlands.

Journal of Virology, August 2003, p. 8470-8480, Vol. 77, No. 15
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.15.8470-8480.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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