Expression of bovine viral diarrhoea virus glycoprotein E2 by bovine herpesvirus-1 from a synthetic ORF and incorporation of E2 into recombinant virions
Expression cassettes containing the codons for the pestivirus E rns signal peptide (Sig) followed by a chemically synthesized ORF that encoded the bovine viral diarrhoea virus (BVDV) strain C86 glycoprotein E2, a class I membrane glycoprotein, were constructed with and without a chimeric intron sequence immediately upstream of the translation start codon, and incorporated into the genome of bovine herpesvirus-1 (BHV-1). The resulting recombinants, BHV- 1/SigE2syn and BHV-1/SigE2syn-intron, expressed comparable quantities of glycoprotein E2, and Northern blot hybridizations indicated that the presence of the intron did not increase significantly the steady-state levels of transcripts encompassing the SigE2syn ORF. In BHV-1/SigE2syn- infected cells, the 54 kDa E2 glycoprotein formed a dimer with an apparent molecular mass of 94 kDa, which was further modified to a 101 kDa form found in the envelope of recombinant virus particles. Penetration kinetics and single-step growth curves indicated that the incorporation of the BVDV E2 glycoprotein in the BHV-1 envelope, which apparently did not require BHV-1-specific signals, interfered with entry into target cells and egress of progeny virions. These results demonstrate that a pestivirus glycoprotein can be expressed efficiently by BHV-1 and incorporated into the viral envelope. BHV-1 thus represents a promising tool for the development of efficacious live and inactivated BHV-1-based vector vaccines.
BecherP.,
OrlichM.,
ShannonA. D.,
HornerG.,
KönigM.,
ThielH.-J.1997; Phylogenetic analysis of pestiviruses from domestic and wild ruminants. Journal of General Virology 78:1357–1366
BühlerB.,
KeilG. M.,
WeilandF.,
KoszinowskiU. H.1990; Characterization of the murine cytomegalovirus early transcription unit e1 that is induced by immediate-early proteins. Journal of Virology 64:1907–1919
EngelsM.,
GiulianiC.,
WildP.,
BeckT. M.,
LoepfeE.,
WylerR.1986; The genome of bovine herpesvirus 1 (BHV-1) strains exhibiting a neuropathogenic potential compared to known BHV-1 strains by restriction site mapping and cross- hybridization. Virus Research 6:57–73
FehlerF.,
HerrmannJ. M.,
SaalmüllerA.,
MettenleiterT. C.,
KeilG. M.1992; Glycoprotein IV of bovine herpesvirus 1-expressing cell line complements and rescues a conditionally lethal viral mutant. Journal of Virology 66:831–839
KeilG. M.,
FibiM. R.,
KoszinowskiU. H.1985; Characterization of the major immediate-early polypeptides encoded by murine cytomegalovirus. Journal of Virology 54:422–428
KeilG. M.,
Ebeling-KeilA.,
KoszinowskiU. H.1987; Sequence and structural organization of murine cytomegalovirus immediate-early gene 1. Journal of Virology 61:1901–1908
KeilG. M.,
EngelhardtT.,
KargerA.,
EnzM.1996; Bovine herpesvirus 1 Us open reading frame 4 encodes a glycoproteoglycan. Journal of Virology 70:3032–3038
KitS.,
KitM.,
DiMarchiR. D.,
LittleS. P.,
GaleC.1991b; Modified-live infectious bovine rhinotracheitis virus vaccine expressing monomer and dimer forms of foot-and-mouth disease capsid protein epitopes on surface of hybrid virus particles. Archives of Virology 120:1–17
KühnleG.,
CollinsR. A.,
ScottJ. E.,
KeilG. M.1996; Bovine interleukins 2 and 4 expressed in recombinant bovine herpesvirus 1 are biologically active secreted glycoproteins. Journal of General Virology 77:2231–2240
KühnleG.,
HeinzeA.,
SchmittJ.,
GiesowK.,
TaylorG.,
MorrisonI.,
RijsewijkF. A. M.,
van OirschotJ. T.,
KeilG. M.1998; The class II membrane glycoprotein G of bovine respiratory syncytial virus, expressed from a synthetic open reading frame, is incorporated into virions of recombinant bovine herpesvirus 1. Journal of Virology 72:3804–3811
Leung-TackP.,
AudonnetJ. C.,
RiviereM.1994; The complete DNA sequence and the genetic organization of the short unique region (US) of the bovine herpesvirus type 1 (ST strain). Virology 199:409–421
McLauchlanJ.,
GaffneyD.,
WhittonJ. L.,
ClementsJ. B.1985; The consensus sequence YGTGTTYY located downstream from the AATAAA signal is required for efficient formation of mRNA 3′ termini. Nucleic Acids Research 13:1347–1368
MayfieldJ. E.,
GoodP. J.,
VanOortH. J.,
CampbellA. R.,
ReedD. E.1983; Cloning and cleavage site mapping of DNA from bovine herpesvirus 1 (Cooper strain. Journal of Virology 47:259–264
MesserleM.,
KeilG. M.,
KoszinowskiU. H.1991; Structure and expression of murine cytomegalovirus immediate-early gene 2. Journal of Virology 65:1638–1643
OtsukaH.,
XuanX.1996; Construction of bovine herpesvirus-1 (BHV-1) recombinants which express pseudorabies virus (PRV) glycoproteins gB, gC, gD, and gE. Archives of Virology 141:57–71
PetitclercD.,
AttalJ.,
TheronM. C.,
BearzottiM.,
BolifraudP.,
KannG.,
StinnakreM. G.,
PointuH.,
PuissantC.,
HoudebineL. M.1995; The effect of various introns and transcription terminators on the efficiency of expression vectors in various cultured cell lines and in the mammary gland of transgenic mice. Journal of Biotechnology 40:169–178
RaggoC.,
FitzpatrickD. R.,
BabiukL. A.,
LiangX.1996; Expression of bovine interleukin-1 beta in a bovine herpesvirus-1 vector: in vitro analysis. Virology 221:78–86
SambrookJ.,
FritschE. F.,
ManiatisT.1989Molecular Cloning: A Laboratory Manual, 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
SchmittJ.,
KeilG. M.1996; Identification and characterization of the bovine herpesvirus 1 UL7 gene and gene product which are not essential for virus replication in cell culture. Journal of Virology 70:1091–1099
SchrijverR. S.,
LangedijkJ. P. M.,
KeilG. M.,
MiddelW. G. J.,
Maris-VeldhuisM.,
Van OirschotJ. T.,
RijsewijkF. A. M.1997; Immunization of cattle with a BHV1 vector vaccine or a DNA vaccine both coding for the G protein of BRSV. Vaccine 15:1908–1916
SchröderC.,
LindeG.,
FehlerF.,
KeilG. M.1997; From essential to beneficial: glycoprotein D loses importance for replication of bovine herpesvirus 1 in cell culture. Journal of Virology 71:25–33
ShiuJ. S.,
LiuS. T.,
ChangT. J.,
HoW. C.,
LaiS. S.,
ChangY. S.1997; The presence of RNA splicing signals in the cDNA construct of the E2 gene of classical swine fever virus affected its expression. Journal of Virological Methods 69:223–230
TakashimaY.,
TamuraH.,
XuanX.,
OtsukaH.1999; Identification of the US3 product of BHV-1 as a protein kinase and characterization of BHV-1 mutants of the US3 gene. Virus Research 59:23–34
TaylorG.,
RijsewijkF. A. M.,
ThomasL. H.,
WyldS. G.,
GaddumR. M.,
CookR. S.,
MorrisonW. I.,
HensenE.,
van OirschotJ. T.,
KeilG.1998; Resistance to bovine respiratory syncytial virus (BRSV) induced in calves by a recombinant bovine herpesvirus-1 expressing the attachment glycoprotein of BRSV. Journal of General Virology 79:1759–1767
TikooS. K.,
FitzpatrickD. R.,
BabiukL. A.,
ZambT. J.1990; Molecular cloning, sequencing, and expression of functional bovine herpesvirus 1 glycoprotein gIV in transfected bovine cells. Journal of Virology 64:5132–5142
van OirschotJ. T.,
KaashoekM. J.,
RijsewijkF. A. M.,
StegemanJ. A.1996; The use of marker vaccines in eradication of herpesviruses. Journal of Biotechnology 44:75–81
van ZijlM.,
WensvoortG.,
de KluyverE.,
HulstM.,
van der GuldenH.,
GielkensA.,
BernsA.,
MoormannR.1991; Live attenuated pseudorabies virus expressing envelope glycoprotein E1 of hog cholera virus protects swine against both pseudorabies and hog cholera. Journal of Virology 65:2761–2765
WeilandE.,
ThielH.-J.,
HessG.,
WeilandF.1989; Development of monoclonal neutralizing antibodies against bovine viral diarrhea virus after pretreatment of mice with normal bovine cells and cyclophosphamide. Journal of Virological Methods 24:237–243
WeilandE.,
StarkR.,
HaasB.,
RümenapfT.,
MeyersG.,
ThielH.-J.1990; Pestivirus glycoprotein which induces neutralizing antibodies forms part of a disulfide-linked heterodimer. Journal of Virology 64:3563–3569
WhitbeckJ. C.,
KnappA. C.,
EnquistL. W.,
LawrenceW. C.,
BelloL. J.1996; Synthesis, processing, and oligomerization of bovine herpesvirus 1 gE and gI membrane proteins. Journal of Virology 70:7878–7884
WylerR.,
EngelsM.,
SchwyzerM.1989; Infectious bovine rhinotracheitis/vulvo-vaginitis (BHV-1).. In Herpesvirus Diseases of Cattle, Horses and Pigs: Developments in Veterinary Virology pp. 1–72 Edited by
WittmannG.
Boston: Kluwer;
Expression of bovine viral diarrhoea virus glycoprotein E2 by bovine herpesvirus-1 from a synthetic ORF and incorporation of E2 into recombinant virions