Construction and characterization of E3-deleted bovine adenovirus type 3 expressing full-length and truncated form of bovine herpesvirus type 1 glycoprotein gD.
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.
Veterinary Infectious Disease Organization, University of Saskatchewan, Saskatoon, Saskatchewan, S7N 5E3, Canada.
A N
AN
Zakhartchouk
P S
PS
Reddy
M
M
Baxi
M E
ME
Baca-Estrada
M
M
Mehtali
L A
LA
Babiuk
S K
SK
Tikoo
D017503:Adenovirus E3 Proteins / Q000235:genetics; D000281:Administration, Intranasal; D000818:Animals; D000914:Antibodies, Viral / Q000032:analysis / Q000097:blood; D000956:Antigens, Viral / Q000032:analysis; D002417:Cattle; D002460:Cell Line; D003561:Cytarabine / Q000494:pharmacology; D004274:DNA, Recombinant; D004279:DNA, Viral / Q000032:analysis / Q000096:biosynthesis; D017353:Gene Deletion; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics*; D007242:Herpesvirus 1, Bovine / Q000235:genetics / Q000276:immunology*; D008168:Lung / Q000276:immunology; D017921:Mastadenovirus / Q000235:genetics* / Q000276:immunology; D009297:Nasal Mucosa / Q000276:immunology; D019384:Nucleic Acid Synthesis Inhibitors / Q000494:pharmacology; D016205:Sigmodontinae; D014611:Vaccination; D019444:Vaccines, DNA / Q000008:administration & dosage / Q000235:genetics* / Q000276:immunology; D014764:Viral Proteins / Q000235:genetics* / Q000276:immunology; D014765:Viral Vaccines / Q000008:administration & dosage / Q000235:genetics* / Q000276:immunology
D017503:Adenovirus E3 Proteins; D000914:Antibodies, Viral; D000956:Antigens, Viral; D004274:DNA, Recombinant; D004279:DNA, Viral; D019384:Nucleic Acid Synthesis Inhibitors; D019444:Vaccines, DNA; D014764:Viral Proteins; D014765:Viral Vaccines; C046998:bovine herpesvirus type-1 glycoproteins; D003561:Cytarabine