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 S 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. Immunofluor escence microscopy also showed that rE2 was transported to the plasma membr ane of infected cells and analysis of purified particles demonstrated that dimeric rE2 was incorporated into VSV-E2 virions in approximately 1:10 rati o with respect to the G glycoprotein. BALB/c mice inoculated intranasally w ith VSV-E2 doses of up to 10(7) plaque forming units (pfu) showed no sympto ms of viral-induced disease and developed a specific BVDV neutralizing resp onse that lasted for at least 180 days post inoculation. (C) 2000 Published by Elsevier Science B.V.