Expression and Immunogenicity of the Spike Glycoprotein of Porcine Respiratory Coronavirus Encoded in the E3 Region of Adenovirus [chapter]

P. Callebaut, M. Pensaert
1995 Advances in Experimental Medicine and Biology  
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 to an amount of approximately 33 J.lg per 10 6 cells, as determined by ELISA. The antigen was cell-associated except in the late phase of the infection, when a low amount (4 J.lg per 10 6
more » ... was released in the culture supernatant. The cell-associated antigen consisted of 2 polypeptides of 160 K and 175 K, respectively. The 160 K polypeptide comigrated with the authentic S' precursor from PRCV-infected cells. The 175 K polypeptide had the same mobility as the authentic mature S protein from PRCV-infected cells and from PRCV released in the supernatant. The extracellular recombinant antigen corresponded with the 175 K mature protein. Immunofluorescent staining gave evidence that some recombinant S protein was exposed on the cell surface; it also showed that the protein was recognized by conformation-specific anti-S monoclonal antibodies. Piglets, immunized oronasally with the recombinant adenovirus vector developed PRCV-neutralizing serum antibodies and were partially protected against PRCV -challenge.
doi:10.1007/978-1-4615-1899-0_42 fatcat:srwumdtqh5ep5imbxnexlweqle