Expression of bovine herpesvirus 1 glycoprotein gIV by recombinant baculovirus and analysis of its immunogenic properties
| Autor: | S. van Drunen Littel-van den Hurk, J.V. van den Hurk, Timothy J. Zamb, Lorne A. Babiuk, Richard Harland, David Fitzpatrick, Michael D. Parker, Manuel Campos |
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| Jazyk: | angličtina |
| Rok vydání: | 1991 |
| Předmět: |
Genes
Viral medicine.drug_class viruses Recombinant Fusion Proteins Immunology Genetic Vectors Fluorescent Antibody Technique Sf9 Enzyme-Linked Immunosorbent Assay Insect Viruses Biology Recombinant virus Monoclonal antibody Transfection Microbiology Epitope law.invention Cell Line Viral Proteins law Virology medicine Animals Neutralizing antibody Herpesvirus 1 Bovine Recombination Genetic fungi biology.organism_classification Flow Cytometry Molecular biology Bovine herpesvirus 1 Kinetics Polyclonal antibodies Insect Science Recombinant DNA biology.protein Cattle Electrophoresis Polyacrylamide Gel Research Article |
| Popis: | The gene encoding the gIV glycoprotein of bovine herpesvirus 1 has been inserted into the genome of Autographa californica baculovirus in lieu of the coding region of the A. californica baculovirus polyhedrin gene. Recombinant protein was identified by its reactivity with gIV-specific monoclonal antibodies and expressed at high levels (about 85 micrograms per 2.5 x 10(6) cells) in Spodoptera frugiperda (SF9) cells. The recombinant glycoprotein had an apparent molecular mass of 63 kDa, indicating that it was incompletely glycosylated. However, it was transported to and expressed on the cell surface of infected SF9 cells. Furthermore, reactivity with polyclonal and monoclonal antibodies specific for gIV suggested that most epitopes were functionally unaltered on the recombinant gIV. Immunization of cattle with recombinant gIV in crude, partially purified, or pure form resulted in the induction of neutralizing antibodies to BHV-1, which were reactive with authentic gIV. However, the neutralizing antibody titers were lower than those elicited by an equivalent amount of affinity-purified authentic gIV, which appeared to be mainly due to reduced recognition of one of the neutralizing antigenic domains of gIV, designated domain I. The potential use of this recombinant gIV glycoprotein as a vaccine to bovine herpesvirus 1 infection in cattle is discussed. |
| Databáze: | OpenAIRE |
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