Assembly of Replication-Incompetent African Horse Sickness Virus Particles: Rational Design of Vaccines for All Serotypes.
Abstract: African horse sickness virus (AHSV), an orbivirus in the Reoviridae family with nine different serotypes, causes devastating disease in equids. The virion particle is composed of seven proteins organized in three concentric layers, an outer layer made of VP2 and VP5, a middle layer made of VP7, and inner layer made of VP3 that encloses a replicase complex of VP1, VP4, and VP6 and a genome of 10 double-stranded RNA segments. In this study, we sought to develop highly efficacious candidate vaccines against all AHSV serotypes, taking into account not only immunogenic and safety properties but also virus productivity and stability parameters, which are essential criteria for vaccine candidates. To achieve this goal, we first established a highly efficient reverse genetics (RG) system for AHSV serotype 1 (AHSV1) and, subsequently, a VP6-defective AHSV1 strain in combination with in trans complementation of VP6. This was then used to generate defective particles of all nine serotypes, which required the exchange of two to five RNA segments to achieve equivalent titers of particles. All reassortant-defective viruses could be amplified and propagated to high titers in cells complemented with VP6 but were totally incompetent in any other cells. Furthermore, these replication-incompetent AHSV particles were demonstrated to be highly protective against homologous virulent virus challenges in type I interferon receptor (IFNAR)-knockout mice. Thus, these defective viruses have the potential to be used for the development of safe and stable vaccine candidates. The RG system also provides a powerful tool for the study of the role of individual AHSV proteins in virus assembly, morphogenesis, and pathogenesis. African horse sickness virus is transmitted by biting midges and causes African horse sickness in equids, with mortality reaching up to 95% in naive horses. Therefore, the development of efficient vaccines is extremely important due to major economic losses in the equine industry. Through the establishment of a highly efficient RG system, replication-deficient viruses of all nine AHSV serotypes were generated. These defective viruses achieved high titers in a cell line complemented with VP6 but failed to propagate in wild-type mammalian or insect cells. Importantly, these candidate vaccine strains showed strong protective efficacy against AHSV infection in an IFNAR(-/-) mouse model.
Copyright © 2016 Lulla et al.
Publication Date: 2016-07-27 PubMed ID: 27279609PubMed Central: PMC4984648DOI: 10.1128/JVI.00548-16Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research article discusses the development of a vaccine for all serotypes of the African Horse Sickness Virus (AHSV), using a highly efficient reverse genetics system and replication-incompetent AHSV particles, which proved to be highly protective against challenges from the virus.
Objective
The objective of the study was to develop a vaccine against all AHSV serotypes. This was prompted by the devastating impact of the disease in equids, with mortality rates of up to 95% in naive horses. The researchers considered not just the immunogenic and safety properties of the vaccine candidates, but also their productivity and stability parameters.
Methodology
- The researchers first established a highly effective reverse genetics (RG) system for AHSV serotype 1 (AHSV1).
- A VP6-defective AHSV1 strain was developed, using a method of in trans complementation of VP6.
- To generate defective particles of all nine serotypes, an exchange of two to five RNA segments was necessary. High titers of particles were achievable through this method.
- All defective viruses produced could be amplified and propagated in cells complemented with VP6, but were not competent in any other cells.
Findings
- The replication-incompetent AHSV particles were proven highly protective against challenges from the virulent virus, in type I interferon receptor (IFNAR)-knockout mice.
- The researchers concluded that these defective viruses could be used in the formulation of safe and stable vaccine candidates.
- Moreover, the RG system used offers a powerful tool for studying the role of individual AHSV proteins in virus assembly, morphogenesis, and pathogenesis.
- The defective viruses achieved high titers in a cell line complemented with VP6 but didn’t propagate in wild-type mammalian or insect cells.
- The potential vaccine strains demonstrated strong protective efficacy against AHSV infection in an IFNAR(-/-) mouse model.
Implications
AHSV is a widespread disease causing significant morbidity and mortality in horses. The development of a safe, effective, and stable vaccine against all serotypes of AHSV could prevent the devastating economic losses incurred by the equine industry due to the disease. The system used for development also provides a new method for studying the role and behavior of virus proteins.
Cite This Article
APA
Lulla V, Lulla A, Wernike K, Aebischer A, Beer M, Roy P.
(2016).
Assembly of Replication-Incompetent African Horse Sickness Virus Particles: Rational Design of Vaccines for All Serotypes.
J Virol, 90(16), 7405-7414.
https://doi.org/10.1128/JVI.00548-16 Publication
Researcher Affiliations
- Department of Pathogen Molecular Biology, Faculty of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine, London, United Kingdom.
- Department of Pathogen Molecular Biology, Faculty of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine, London, United Kingdom.
- Institute of Diagnostic Virology, Friedrich-Loeffler Institut, Greifswald, Germany.
- Institute of Diagnostic Virology, Friedrich-Loeffler Institut, Greifswald, Germany.
- Institute of Diagnostic Virology, Friedrich-Loeffler Institut, Greifswald, Germany.
- Department of Pathogen Molecular Biology, Faculty of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine, London, United Kingdom polly.roy@lshtm.ac.uk.
MeSH Terms
- African Horse Sickness / prevention & control
- African Horse Sickness Virus / genetics
- African Horse Sickness Virus / immunology
- African Horse Sickness Virus / physiology
- Animals
- Defective Viruses / genetics
- Defective Viruses / immunology
- Defective Viruses / physiology
- Disease Models, Animal
- Gene Deletion
- Mice
- Mice, Knockout
- Reverse Genetics
- Serogroup
- Viral Vaccines / administration & dosage
- Viral Vaccines / immunology
- Viral Vaccines / metabolism
- Virion / metabolism
- Virus Assembly
- Virus Replication
Grant Funding
- BB/K015168/1 / Biotechnology and Biological Sciences Research Council
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