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-SD...

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Bibliographic Details
Published in:Virus research 2000-08, Vol.69 (1), p.3-15
Main Authors: Grigera, P.R, Marzocca, M.P, Capozzo, A.V.E, Buonocore, L, Donis, R.O, Rose, J.K
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Animals
Antibodies, Viral - biosynthesis
Antibodies, Viral - blood
Base Sequence
BHK21 cell expression
Bovine viral diarrhea virus
BVDV-E2 glycoprotein
Cattle
Cell Line
Chimera - genetics
Chimera - immunology
Cricetinae
Diarrhea Viruses, Bovine Viral - genetics
Diarrhea Viruses, Bovine Viral - immunology
DNA, Recombinant - genetics
Female
Glycoprotein E2
Live vaccines
Membrane Glycoproteins
Mice
Mice, Inbred BALB C
Neutralization Tests
Recombinant Proteins - genetics
Recombinant Proteins - immunology
Recombinant VSV
Vaccines, Synthetic - genetics
Vesicular stomatitis Indiana virus - genetics
Viral Envelope Proteins - genetics
Viral Envelope Proteins - immunology
Viral Vaccines - genetics
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Summary: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.
ISSN:0168-1702
1872-7492
DOI:10.1016/S0168-1702(00)00164-7