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Development and characterization of an infectious cDNA clone of the virulent Bucyrus strain of Equine arteritis virus.
Abstract: Strains of Equine arteritis virus (EAV) differ in the severity of the disease that they induce in horses. Infectious cDNA clones are potentially useful for identification of genetic determinants of EAV virulence; to date, two clones have been derived from a cell culture-adapted variant of the original (Bucyrus) isolate of EAV, and it has previously been shown that recombinant virus derived from one of these (rEAV030) is attenuated in horses. A complete cDNA copy of the genome of the virulent Bucyrus strain of EAV has now been assembled into a plasmid vector. In contrast to rEAV030, recombinant progeny virus derived from this clone caused severe disease in horses, characterized by pyrexia, oedema, leukopenia, high-titre viraemia and substantial nasal shedding of virus. The availability of infectious cDNA clones that produce recombinant viruses of different virulence to horses will facilitate characterization of the virulence determinants of EAV through reverse genetics.
Publication Date: 2007-02-28 PubMed ID: 17325365DOI: 10.1099/vir.0.82415-0Google Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
- Research Support
- Non-U.S. Gov't
- Research Support
- U.S. Gov't
- Non-P.H.S.
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
This research focuses on the development of a comprehensive cDNA copy of the genome for the virulent Bucyrus strain of Equine arteritis virus (EAV). The researchers show that the strain, when replicated in horses, led to severe disease. This project aids in identifying the virulence determinants of this specific strain.
Objectives and Methodology
- The study’s primary goal was to create an infectious cDNA clone of the virulent Bucyrus strain of Equine arteritis virus. cDNA, or complementary DNA, is a form of DNA synthesized from a mature strand of messenger RNA by the enzyme reverse transcriptase. Having a cDNA clone allows researchers to study and manipulate the virus in a laboratory setting.
- Previous efforts in this field have resulted in the creation of two clones derived from a less harmful, cell culture-adapted variant of the original Bucyrus isolate of EAV. The new research aimed at creating a clone from the more harmful strain of the virus to provide more insight into the genetic determinants of EAV virulence.
Main Findings
- The researchers successfully developed a comprehensive cDNA copy of the genome of the virulent Bucyrus EAV strain and assembled this into a plasmid vector.
- Unlike the recombinant virus derived from previous cDNA clones (rEAV030), which was attenuated (reduced in virulence) in horses, the recombinant virus created using this new cDNA clone caused severe disease in horses.
- The disease was characterized by high fever (pyrexia), swelling (oedema), a decrease in white blood cells (leukopenia), high viral load (high-titre viraemia), and a substantial nasal shedding of the virus.
Significance annd Future Reseach
- The successful production of this infectious cDNA clone of a virulent EAV strain allows for detailed study of the determinants of virulence in this virus.
- With cDNA clones representing different virulence levels in horses, the researchers can conduct more comprehensive studies through reverse genetics, a method used to identify the function of a gene by analyzing the phenotypic effects of specific genes sequences.
- This research marks significant progress towards the characterization of the virulence determinants of EAV strains, which could lead to the development of targeted therapies or preventive measures against the disease.
Cite This Article
APA
Balasuriya UBR, Snijder EJ, Heidner HW, Zhang J, Zevenhoven-Dobbe JC, Boone JD, McCollum WH, Timoney PJ, MacLachlan NJ.
(2007).
Development and characterization of an infectious cDNA clone of the virulent Bucyrus strain of Equine arteritis virus.
J Gen Virol, 88(Pt 3), 918-924.
https://doi.org/10.1099/vir.0.82415-0 Publication
Researcher Affiliations
- Department of Pathology, Microbiology and Immunology, School of Veterinary Medicine, University of California, Davis, CA 95616, USA.
- Molecular Virology Laboratory, Department of Medical Microbiology, Leiden University Medical Center, Leiden, The Netherlands.
- Department of Biology, University of Texas at San Antonio, TX 78249, USA.
- Department of Veterinary Science, Gluck Equine Research Center, University of Kentucky, Lexington, KY 40546, USA.
- Molecular Virology Laboratory, Department of Medical Microbiology, Leiden University Medical Center, Leiden, The Netherlands.
- Department of Pathology, Microbiology and Immunology, School of Veterinary Medicine, University of California, Davis, CA 95616, USA.
- Department of Veterinary Science, Gluck Equine Research Center, University of Kentucky, Lexington, KY 40546, USA.
- Department of Veterinary Science, Gluck Equine Research Center, University of Kentucky, Lexington, KY 40546, USA.
- Department of Pathology, Microbiology and Immunology, School of Veterinary Medicine, University of California, Davis, CA 95616, USA.
MeSH Terms
- Animals
- Arterivirus Infections / physiopathology
- Arterivirus Infections / veterinary
- Arterivirus Infections / virology
- DNA, Complementary
- DNA, Viral / chemistry
- DNA, Viral / genetics
- Equartevirus / genetics
- Equartevirus / pathogenicity
- Equartevirus / physiology
- Genetic Vectors
- Genome, Viral
- Horse Diseases / physiopathology
- Horse Diseases / virology
- Horses
- Molecular Sequence Data
- Plasmids / genetics
- Viremia
- Virus Shedding
Citations
This article has been cited 21 times.- Thieulent CJ, Sarkar S, Carossino M, Bhowmik M, Zhu H, Balasuriya UBR. Cell Surface Vimentin Is an Attachment Factor That Facilitates Equine Arteritis Virus Infection In Vitro. Viruses 2026 Jan 15;18(1).
- Thieulent CJ, Carossino M, Balasuriya UBR, Graves K, Bailey E, Eberth J, Canisso IF, Andrews FM, Keowen ML, Go YY. Development of a TaqMan(®) Allelic Discrimination qPCR Assay for Rapid Detection of Equine CXCL16 Allelic Variants Associated With the Establishment of Long-Term Equine Arteritis Virus Carrier State in Stallions. Front Genet 2022;13:871875.
- Chaudhari J, Nguyen TN, Vu HLX. Identification of Cryptic Promoter Activity in cDNA Sequences Corresponding to PRRSV 5' Untranslated Region and Transcription Regulatory Sequences. Viruses 2022 Feb 15;14(2).
- Carossino M, Dini P, Kalbfleisch TS, Loynachan AT, Canisso IF, Cook RF, Timoney PJ, Balasuriya UBR. Equine arteritis virus long-term persistence is orchestrated by CD8+ T lymphocyte transcription factors, inhibitory receptors, and the CXCL16/CXCR6 axis. PLoS Pathog 2019 Jul;15(7):e1007950.
- Nam B, Mekuria Z, Carossino M, Li G, Zheng Y, Zhang J, Cook RF, Shuck KM, Campos JR, Squires EL, Troedsson MHT, Timoney PJ, Balasuriya UBR. Intrahost Selection Pressure Drives Equine Arteritis Virus Evolution during Persistent Infection in the Stallion Reproductive Tract. J Virol 2019 Jun 15;93(12).
- Carossino M, Dini P, Kalbfleisch TS, Loynachan AT, Canisso IF, Shuck KM, Timoney PJ, Cook RF, Balasuriya UBR. Downregulation of MicroRNA eca-mir-128 in Seminal Exosomes and Enhanced Expression of CXCL16 in the Stallion Reproductive Tract Are Associated with Long-Term Persistence of Equine Arteritis Virus. J Virol 2018 May 1;92(9).
- Carossino M, Wagner B, Loynachan AT, Cook RF, Canisso IF, Chelvarajan L, Edwards CL, Nam B, Timoney JF, Timoney PJ, Balasuriya UBR. Equine Arteritis Virus Elicits a Mucosal Antibody Response in the Reproductive Tract of Persistently Infected Stallions. Clin Vaccine Immunol 2017 Oct;24(10).
- Carossino M, Loynachan AT, Canisso IF, Cook RF, Campos JR, Nam B, Go YY, Squires EL, Troedsson MHT, Swerczek T, Del Piero F, Bailey E, Timoney PJ, Balasuriya UBR. Equine Arteritis Virus Has Specific Tropism for Stromal Cells and CD8(+) T and CD21(+) B Lymphocytes but Not for Glandular Epithelium at the Primary Site of Persistent Infection in the Stallion Reproductive Tract. J Virol 2017 Jul 1;91(13).
- Gulyaeva A, Dunowska M, Hoogendoorn E, Giles J, Samborskiy D, Gorbalenya AE. Domain Organization and Evolution of the Highly Divergent 5' Coding Region of Genomes of Arteriviruses, Including the Novel Possum Nidovirus. J Virol 2017 Mar 15;91(6).
- Sarkar S, Bailey E, Go YY, Cook RF, Kalbfleisch T, Eberth J, Chelvarajan RL, Shuck KM, Artiushin S, Timoney PJ, Balasuriya UB. Allelic Variation in CXCL16 Determines CD3+ T Lymphocyte Susceptibility to Equine Arteritis Virus Infection and Establishment of Long-Term Carrier State in the Stallion. PLoS Genet 2016 Dec;12(12):e1006467.
- Sarkar S, Chelvarajan L, Go YY, Cook F, Artiushin S, Mondal S, Anderson K, Eberth J, Timoney PJ, Kalbfleisch TS, Bailey E, Balasuriya UB. Equine Arteritis Virus Uses Equine CXCL16 as an Entry Receptor. J Virol 2016 Jan 13;90(7):3366-84.
- Vatter HA, Di H, Donaldson EF, Baric RS, Brinton MA. Each of the eight simian hemorrhagic fever virus minor structural proteins is functionally important. Virology 2014 Aug;462-463:351-62.
- Go YY, Li Y, Chen Z, Han M, Yoo D, Fang Y, Balasuriya UB. Equine arteritis virus does not induce interferon production in equine endothelial cells: identification of nonstructural protein 1 as a main interferon antagonist. Biomed Res Int 2014;2014:420658.
- Balasuriya UB, Zhang J, Go YY, MacLachlan NJ. Experiences with infectious cDNA clones of equine arteritis virus: lessons learned and insights gained. Virology 2014 Aug;462-463:388-403.
- Balasuriya UB, Go YY, MacLachlan NJ. Equine arteritis virus. Vet Microbiol 2013 Nov 29;167(1-2):93-122.
- Zhang J, Go YY, Huang CM, Meade BJ, Lu Z, Snijder EJ, Timoney PJ, Balasuriya UB. Development and characterization of an infectious cDNA clone of the modified live virus vaccine strain of equine arteritis virus. Clin Vaccine Immunol 2012 Aug;19(8):1312-21.
- Firth AE, Zevenhoven-Dobbe JC, Wills NM, Go YY, Balasuriya UBR, Atkins JF, Snijder EJ, Posthuma CC. Discovery of a small arterivirus gene that overlaps the GP5 coding sequence and is important for virus production. J Gen Virol 2011 May;92(Pt 5):1097-1106.
- Go YY, Snijder EJ, Timoney PJ, Balasuriya UB. Characterization of equine humoral antibody response to the nonstructural proteins of equine arteritis virus. Clin Vaccine Immunol 2011 Feb;18(2):268-79.
- Go YY, Zhang J, Timoney PJ, Cook RF, Horohov DW, Balasuriya UB. Complex interactions between the major and minor envelope proteins of equine arteritis virus determine its tropism for equine CD3+ T lymphocytes and CD14+ monocytes. J Virol 2010 May;84(10):4898-911.
- Zhang J, Timoney PJ, MacLachlan NJ, McCollum WH, Balasuriya UB. Persistent equine arteritis virus infection in HeLa cells. J Virol 2008 Sep;82(17):8456-64.
- Go YY, Wong SJ, Branscum AJ, Demarest VL, Shuck KM, Vickers ML, Zhang J, McCollum WH, Timoney PJ, Balasuriya UB. Development of a fluorescent-microsphere immunoassay for detection of antibodies specific to equine arteritis virus and comparison with the virus neutralization test. Clin Vaccine Immunol 2008 Jan;15(1):76-87.
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