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Use of recombinant gp135 to study epitope-specific antibody responses to maedi visna virus
N Carey1, D J Roy, R G Dalziel
1Department of Veterinary Pathology, University of Edinburgh, Summerhall, UK.
Abstract:
The envelope glycoprotein gp135 of the ovine lentivirus maedi visna virus (MVV) is the main target for neutralising antibody in vivo, however little is known about the specific regions of gp135 which elicit this neutralising response. We have used the polymerase chain reaction (PCR) to generate overlapping fragments of the gp135 gene which have been expressed as fusion proteins in the yeast Ty-VLP system. These fusion proteins have been used to analyse the antibody response to gp135 in MVV infected sheep and we are able to identify at least three distinct regions of gp135 to which antibodies are directed. The approach described in this paper provides a rapid and simple method of generating overlapping fusion proteins with which to carry out epitope mapping studies.
Insights
Researchers identified key regions of the maedi visna virus (MVV) envelope glycoprotein gp135 targeted by neutralizing antibodies in sheep. This study utilized polymerase chain reaction (PCR) and yeast expression for rapid epitope mapping.
Area of Science:
- Virology
- Immunology
- Molecular Biology
Background:
- The envelope glycoprotein gp135 of ovine lentivirus maedi visna virus (MVV) is crucial for viral entry and a primary target for neutralizing antibodies.
- However, the specific epitopes on gp135 responsible for eliciting a neutralizing antibody response remain largely uncharacterized.
Purpose of the Study:
- To identify specific regions of the MVV gp135 glycoprotein that are targeted by neutralizing antibodies in infected sheep.
- To develop a rapid and efficient method for epitope mapping of viral glycoproteins.
Main Methods:
- Utilized polymerase chain reaction (PCR) to generate overlapping fragments of the gp135 gene.
- Expressed these fragments as fusion proteins in the yeast Ty-VLP system.
- Analyzed the antibody response in MVV-infected sheep against these fusion proteins.
Main Results:
- Identified at least three distinct regions on the gp135 glycoprotein that elicit antibody responses in MVV-infected sheep.
- Demonstrated the utility of the yeast Ty-VLP system for producing fusion proteins for immunological studies.
Conclusions:
- The study successfully mapped key antigenic regions of the MVV gp135.
- The described method offers a rapid and simple approach for epitope mapping of viral glycoproteins, aiding in vaccine development and diagnostics.