Presence of bovine viral diarrhea virus (BVDV) E2 glycoprotein in VSV recombinant particles and induction of neutralizing BVDV antibodies in mice.
We generated a recombinant vesicular stomatitis virus (VSV-E2) encoding the bovine viral diarrhea virus (BVDV) E2 glycoprotein with the VSV-G protein signal peptide. Infection of BHK21 cells with VSV-E2 induced the synthesis of a recombinant E2 (rE2) that comigrated with authentic BVDV-E2 in PAGE-SDS gels. Non-reducing immunoblots showed that rE2 is a disulfide bond-linked homodimer with at least 10-fold higher avidity for conformation-dependent anti-BVDV-E2 antibodies than its reduced monomeric counterpart. Immunofluorescence microscopy also showed that rE2 was transported to the plasma membrane of infected cells and analysis of purified particles demonstrated that dimeric rE2 was incorporated into VSV-E2 virions in approximately 1:10 ratio with respect to the G glycoprotein. BALB/c mice inoculated intranasally with VSV-E2 doses of up to 10(7) plaque forming units (pfu) showed no symptoms of viral-induced disease and developed a specific BVDV neutralizing response that lasted for at least 180 days post inoculation.
Grigera
P R
PR
Centro de Virologia Animal, Serrano 669, 1414 Capital Federal, Buenos Aires, Argentina.
Marzocca
M P
MP
Capozzo
A V
AV
Buonocore
L
L
Donis
R O
RO
Rose
J K
JK
D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral / Q000096:biosynthesis* / Q000097:blood; D001483:Base Sequence; D002417:Cattle; D002460:Cell Line; D002678:Chimera / Q000235:genetics / Q000276:immunology; D006224:Cricetinae; D004274:DNA, Recombinant / Q000235:genetics; D001908:Diarrhea Viruses, Bovine Viral / Q000235:genetics* / Q000276:immunology*; D005260:Female; D008562:Membrane Glycoproteins; D051379:Mice; D008807:Mice, Inbred BALB C; D009500:Neutralization Tests; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D014614:Vaccines, Synthetic / Q000235:genetics; D014721:Vesicular stomatitis Indiana virus / Q000235:genetics*; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000276:immunology*; D014765:Viral Vaccines / Q000235:genetics
D000914:Antibodies, Viral; D004274:DNA, Recombinant; C028391:G protein, vesicular stomatitis virus; D008562:Membrane Glycoproteins; D011994:Recombinant Proteins; D014614:Vaccines, Synthetic; D014759:Viral Envelope Proteins; D014765:Viral Vaccines; C076551:gp53, bovine viral diarrhea virus