Construction and Characterization of E3-Deleted Bovine Adenovirus Type 3 Expressing Full-Length and Truncated Form of Bovine Herpesvirus Type 1 Glycoprotein gD

Abstract

Using the homologous recombination machinery of E. coli, a 1.245-kb deletion was introduced in the E3 region of bovine adenovirus 3 (BAV3) genomic DNA cloned in a plasmid. Transfection of the restriction enzyme-excised, linear E3-deleted BAV3 genomic DNA into primary fetal bovine retina cells produced infectious virus (BAV3. E3d), suggesting that all the E3-specific open reading frames are nonessential for virus replication in vitro. Using a similar approach, we constructed replication-competent (BAV3.E3gD and BAV3. E3gDt) BAV3 recombinant expressing full-length (gD) or truncated (gDt) glycoprotein of bovine herpes virus 1. Recombinant gD and gDt proteins expressed by BAV3.E3gD and BAV3.E3gDt, respectively, were recognized by gD-specific monoclonal antibodies directed against conformational epitopes, suggesting that antigenicity of recombinant gD and gDt was similar to that of the native gD expressed in bovine herpes virus 1-infected cells. Intranasal immunization of cotton rats induced strong gD- and BAV3-specific IgA and IgG immune responses. These results suggest that replication-competent bovine adenovirus 3-based vectors have potential for the delivery of vaccine antigens to the mucosal surfaces of animals.

Optimization of a DNA Vaccine Against SARS

“…Previously, we have shown that intranasal immunization of cotton rats with recombinant adenovirus expressing a soluble form of the BHV-1 gD protein induced a higher antibody response than immunization with the virus expressing the membrane-anchored form of the protein (Zakhartchouk et al, 1998). However, in the present study, we demonstrated that pcDNA-St induced lower immune responses in vaccinated mice than pcDNA-SS.…”

Prevention of rabies virus infection in dogs by a recombinant canine adenovirus type-2 encoding the rabies virus glycoprotein

“…The E3 region of adenovirus is not essential for viral replication in tissue culture, but it is reported that E3 is involved in the regulation of immune response in vivo [34,35]. The deletion of E3a has been reported to enhance pulmonary lymphocyte infiltration when the deleted virus was administrated intranasally in cotton rats [36].…”

Generation of E3-deleted canine adenovirus type 2 expressing the Gc glycoprotein of Seoul virus by gene insertion or deletion of related terminal region sequences

“…Replication-competent recombinant adenoviruses of several different species have been developed by homologous recombination, with insertion of foreign genes into the non-essential E3 region (Morrison et al, 2002;Zakhartchouk et al, 1998). The present study, however, used a different approach.…”