Expression of bovine herpesvirus 1 glycoprotein gIV by recombinant baculovirus and analysis of its immunogenic properties.
The gene encoding the gIV glycoprotein of bovine herpesvirus 1 has been inserted into the genome of Autographa californica baculovirus in lieu of the coding region of the A. californica baculovirus polyhedrin gene. Recombinant protein was identified by its reactivity with gIV-specific monoclonal antibodies and expressed at high levels (about 85 micrograms per 2.5 x 10(6) cells) in Spodoptera frugiperda (SF9) cells. The recombinant glycoprotein had an apparent molecular mass of 63 kDa, indicating that it was incompletely glycosylated. However, it was transported to and expressed on the cell surface of infected SF9 cells. Furthermore, reactivity with polyclonal and monoclonal antibodies specific for gIV suggested that most epitopes were functionally unaltered on the recombinant gIV. Immunization of cattle with recombinant gIV in crude, partially purified, or pure form resulted in the induction of neutralizing antibodies to BHV-1, which were reactive with authentic gIV. However, the neutralizing antibody titers were lower than those elicited by an equivalent amount of affinity-purified authentic gIV, which appeared to be mainly due to reduced recognition of one of the neutralizing antigenic domains of gIV, designated domain I. The potential use of this recombinant gIV glycoprotein as a vaccine to bovine herpesvirus 1 infection in cattle is discussed.
van Drunen Littel-van den Hurk
S
S
Veterinary Infectious Disease Organization, University of Saskatchewan, Saskatoon, Canada.
Parker
M D
MD
Fitzpatrick
D R
DR
Zamb
T J
TJ
van den Hurk
J V
JV
Campos
M
M
Harland
R
R
Babiuk
L A
LA
D000818:Animals; D002417:Cattle; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D005814:Genes, Viral; D005822:Genetic Vectors; D007242:Herpesvirus 1, Bovine / Q000235:genetics*; D007304:Insect Viruses / Q000235:genetics*; D007700:Kinetics; D011993:Recombinant Fusion Proteins / Q000276:immunology / Q000302:isolation & purification; D011995:Recombination, Genetic; D014162:Transfection; D014764:Viral Proteins / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification
D011993:Recombinant Fusion Proteins; D014764:Viral Proteins; C046998:bovine herpesvirus type-1 glycoproteins