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.
AndrewM. E.,
BoyleD. B.,
WhitefeldP. L.,
LockettL. J.,
AnthonyI. D.,
BellamyA. R.,
BothG. W.1990; The immunogenicity of VP7, a rotavirus antigen resident in the endoplasmic reticulum is enhanced by cell-surface expression. Journal of Virology 64:4776–4783
CallebautP.,
DebouckP.,
PensaertM.1982; Enzyme-linked immunosorbent assay for the detection of the coronavirus–like agent and its antibodies in pigs with porcine epidemic diarrhea. Veterinary Microbiology 7:295–306
CallebautP.,
CorreaI.,
PensaertM.,
JiménezG.,
EnjuanesL.1988; Antigenic differentiation between transmissible gastroenteritis virus of swine and a related porcine respiratory coronavirus. Journal of General Virology 69:1725–1730
CallebautP.,
PensaertM.,
EnjuanesL.1994; Construction of a recombinant adenovirus for the expression of the glycoprotein S antigen of porcine respiratory coronavirus. Advances in Experimental Medicine and Biology 342:469–470
CharltonK. M.,
ArtoisM.,
PrevecL.,
CampbellJ. B.,
CasayG. A.,
WandelerA. I.,
ArmstrongJ.1992; Oral rabies vaccination of skunks and foxes with a recombinant human adenovirus vaccine. Archives of Virology 123:169–179
CorreaI.,
JiménezG.,
SuneC.,
BullidoM. J.,
EnjuanesL.1988; Antigenic structure of the E2 glycoprotein from transmissible gastroenteritis coronavirus. Virus Research 10:77–94
DelmasB.,
GelfiJ.,
LaudeH.1986; Antigenic structure of transmissible gastroenteritis virus. II. Domains in the peplomer glycoprotein. Journal of General Virology 67:1405–1418
GodetM.,
RaschaertD.,
LaudeH.1991; Processing and antigenicity of entire and anchor-free spike glycoprotein S of coronavirus TGEV expressed by recombinant baculovirus. Virology 185:732–740
PensaertM.,
CallebautP.,
VergoteJ.1986; Isolation of a porcine respiratory, non–enteric coronavirus related to transmissible gastroenteritis. Veterinary Quarterly 8:257–261
RasschaertD.,
DuarteM.,
LaudeH.1990; Porcine respiratory coronavirus differs from transmissible gastroenteritis virus by a few genomic deletions. Journal of General Virology 71:2599–2607
SanchezC. M.,
JimenezG.,
LaviadaM. D.,
CorreaL.,
SunéC.,
BullidoM. J.,
GebauerF.,
SmerdouC.,
CallebautP.,
EscribanoJ. M.,
EnjuanesL.1990; Antigenic homology among coronaviruses related to transmissible gastroenteritis virus. Virology 174:410–417
ToL. T.,
BernardS.,
BottreauE.1992; Transmissible gastroenteritis coronavirus: surface antigens induced by virulent and attenuated strains. Research in Virology 143:241–248
TubolyT.,
NagyE.,
DerbyshireJ. B.1993; Potential viral vectors for the stimulation of mucosal antibody responses against enteric antigens in pigs. Research in Veterinary Science 54:345–350
Van ReethK.,
PensaertM.1994; Porcine respiratory coronavirus-mediated interference against influenza virus replication in the respiratory tract of feeder pigs. American Journal of Veterinary Research 55:1275–1281
WesselingJ. G.,
GodekeG.,
SchijnsV. E. C. J.,
PrevecL.,
GrahamF. L.,
HorzinekM. C.,
RottierP. J. M.1993; Mouse hepatitis virus spike and nucleocapsid proteins expressed by adenovirus vectors protect mice against a lethal infection. Journal of General Virology 74:2061–2069
WoldW. S. M.,
GoodingL. R.1991; Region E3 of adenovirus: a cassette of genes involved in host immunosurveillance and virus–cell interactions. Virology 184:1–8