Herpes simplex virus type-1 (HSV-1) amplicon vectors are versatile and useful tools for transferring genes into cells that are capable of stimulating a specific immune response to their expressed antigens. In this work, two HSV-1-derived amplicon vectors were generated. One of these expressed the full-length glycoprotein D (gD) of bovine herpesvirus 1 while the second expressed the truncated form of gD (gDtr) which lacked the trans-membrane region. After evaluating gD expression in the infected cells, the ability of both vectors to induce a specific gD immune response was tested in BALB/c mice that were intramuscularly immunized. Specific serum antibody responses were detected in mice inoculated with both vectors, and the response against truncated gD was higher than the response against full-length gD. These results reinforce previous findings that HSV-1 amplicon vectors can potentially deliver antigens to animals and highlight the prospective use of these vectors for treating infectious bovine rhinotracheitis disease.
Animals ; Antibodies, Viral/blood ; Blotting, Western/veterinary ; Cattle ; Female ; Genetic Vectors/*immunology ; Herpesvirus 1, Bovine/genetics/*immunology ; Herpesvirus 1, Human/genetics/*immunology ; Immunity, Humoral/immunology ; Immunization/methods/veterinary ; Infectious Bovine Rhinotracheitis/*immunology/prevention & control/virology ; Mice ; Mice, Inbred BALB C ; Neutralization Tests/veterinary ; Specific Pathogen-Free Organisms ; Viral Proteins/genetics/*immunology ; Viral Vaccines/immunology

Taking the genome DNA of Infectious Bovine Rhinotracheitis Virus (IBRV) as the template, the gG gene was amplified with PCR and cloned into the T cloning vector pMD18-T. After being identified by restriction digestion and DNA sequencing, the insert was subcloned into the expression vector pGEX-KG. Sodium docecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot assay showed that this gene was expressed as both soluble form and inclusion body by the transformed E. coli BL21 strain (DE3). The fusion protein was purified and used as the coating antigen to develop the indirect Enzyme-Linked Immunosorbent Assay (ELISA). Comparison between this gG-ELISA and commercial IBRV gB-ELISA Kit (IDEXX) was made in the detection of 380 cow serum samples. The results demonstrated an agreement of 92%. By using this novel gG-ELISA, 1248 cow serum samples were tested and the average positive rate of IBRV antibodies for imported cows is 21.7%, while the positive rate ranged greatly from 0.0%-41.5% for Hubei local Chinese Black and White Dairy Cows.
Animals ; Antibodies, Viral ; blood ; immunology ; Antigens, Viral ; genetics ; immunology ; Cattle ; Cloning, Molecular ; Enzyme-Linked Immunosorbent Assay ; methods ; Escherichia coli ; genetics ; metabolism ; Female ; Herpesvirus 1, Bovine ; genetics ; immunology ; Male ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; Sensitivity and Specificity ; Viral Proteins ; biosynthesis ; genetics ; immunology