Rescue of disabled infectious single-cycle (DISC) equine arteritis virus by using complementing cell lines that express minor structural glycoproteins.
Abstract: Equine arteritis virus (EAV) contains seven structural proteins that are all required to produce infectious progeny. Alphavirus-based expression vectors have been generated for each of these proteins to explore the possibilities for their constitutive expression in cell lines. This approach was successful for minor glycoproteins GP(2b), GP(3) and GP(4) and for the E protein. Subsequently, it was demonstrated that cell lines expressing these proteins could rescue EAV mutants that were disabled in the expression of the corresponding gene, resulting in the production of virus particles carrying the mutant genome. This system was particularly efficient for GP(2b)- and GP(4)-knockout mutants. Upon infection of non-complementing cells with these mutants, a self-limiting single cycle of replication was initiated, resulting in the expression of all but one of the viral proteins. These disabled infectious single-cycle (DISC) arteriviruses can also be used to express foreign sequences and are potentially useful in both fundamental research and vaccine development.
Publication Date: 2004-11-24 PubMed ID: 15557244DOI: 10.1099/vir.0.80443-0Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This research explores the use of cell lines expressing certain viral proteins to produce virus particles of the equine arteritis virus (EAV), specifically those that have random gene mutations. The successful implementation of the experiment could provide a basis for further research and potentially the development of vaccines.
Exploring Structural Proteins
- Equine arteritis virus (EAV) contains seven structural proteins – all of which are essential in producing infectious offspring.
- To further understand these proteins, the researchers used alphavirus-based expression vectors – these are a genetic tool used to make cells express a specific gene.
- By applying this to all seven proteins, it was found that minor glycoproteins GP(2b), GP(3) and GP(4) and the E protein were able to be expressed in cell lines.
Complementing Cell Lines
- The study established that cell lines which constitutively expressed these proteins were able to rescue EAV mutants that couldn’t express the corresponding gene.
- These mutants eventually produced virus particles that contained the mutant genome – a particularly successful result was found with the GP(2b) and GP(4) knockout mutants.
Disabled Infectious Single-Cycle (DISC) Arteriviruses
- When these mutated viruses were factored into non-complementing cells, it triggered a limited single cycle of replication, leading to the expression of all other viral proteins, except for one.
- The researchers identified these versions of the virus as disabled infectious single-cycle (DISC) arteriviruses.
- Due to their unique properties, DISC arteriviruses can serve as a good platform for expressing foreign sequences, which is beneficial for fundamental research.
- Besides, DISC arteriviruses contain the potential to be used in vaccine development due to their ability to produce offsprings with a partial viral genome.
Cite This Article
APA
Zevenhoven-Dobbe JC, Greve S, van Tol H, Spaan WJM, Snijder EJ.
(2004).
Rescue of disabled infectious single-cycle (DISC) equine arteritis virus by using complementing cell lines that express minor structural glycoproteins.
J Gen Virol, 85(Pt 12), 3709-3714.
https://doi.org/10.1099/vir.0.80443-0 Publication
Researcher Affiliations
- Molecular Virology Laboratory, Department of Medical Microbiology, Center of Infectious Diseases, Leiden University Medical Center, LUMC E4-P, Room P4-26, PO Box 9600, 2300 RC Leiden, The Netherlands.
- Molecular Virology Laboratory, Department of Medical Microbiology, Center of Infectious Diseases, Leiden University Medical Center, LUMC E4-P, Room P4-26, PO Box 9600, 2300 RC Leiden, The Netherlands.
- Molecular Virology Laboratory, Department of Medical Microbiology, Center of Infectious Diseases, Leiden University Medical Center, LUMC E4-P, Room P4-26, PO Box 9600, 2300 RC Leiden, The Netherlands.
- Molecular Virology Laboratory, Department of Medical Microbiology, Center of Infectious Diseases, Leiden University Medical Center, LUMC E4-P, Room P4-26, PO Box 9600, 2300 RC Leiden, The Netherlands.
- Molecular Virology Laboratory, Department of Medical Microbiology, Center of Infectious Diseases, Leiden University Medical Center, LUMC E4-P, Room P4-26, PO Box 9600, 2300 RC Leiden, The Netherlands.
MeSH Terms
- Animals
- Cell Line
- Cricetinae
- Defective Viruses / physiology
- Equartevirus / physiology
- Viral Structural Proteins / physiology
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