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1 Laboratory of Veterinary Virology, Faculty of Veterinary Medicine, University of Ghent, Salisburylaan 133, B-9820 Merelbeke, Belgium
and2 Centro Nacional de Biotecnologia, CSIC, Campus Universidad Autónoma, Canto Blanco, E-28049 Madrid, Spain
The full-length spike (S) gene of porcine respiratory coronavirus (PRCV) was inserted into the genome of human adenovirus type 5 downstream of the early transcription region 3 promoter. The recombinant virus replicated in cultures of the swine testicle ST cell line and directed the synthesis of S antigen with a maximum yield of approximately 26 µg per 106 cells. The antigen was cell-associated except in the late phase of the infection, when a small amount (3.5 µg per 106 cells) was released. The cell-associated antigen consisted of polypeptides of molecular mass 160 kDa and 175 kDa, comigrating with the authentic precursor S' and the mature S protein of PRCV, respectively. The extracellular recombinant antigen corresponded to the 175 kDa mature protein. Some recombinant S protein was exposed on the cell surface and was recognized by neutralization-mediating anti-S monoclonal antibodies. Piglets, inoculated oronasally with the recombinant adenovirus vector developed PRCV-neutralizing serum antibodies and were partially protected against PRCV challenge, demonstrating the potential of live adenovirus as vaccine vector.
* Author for correspondence. Fax +32 9 264 74 95.
Received 18 May 1995;
accepted 29 September 1995.
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