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Virology1999; 260(1); 201-208; doi: 10.1006/viro.1999.9817

Equine arteritis virus derived from an infectious cDNA clone is attenuated and genetically stable in infected stallions.

Abstract: Virus derived from an infectious cDNA clone of equine arteritis virus (EAV030H) was intranasally inoculated into two stallions, neither of which subsequently developed clinical manifestations of equine viral arteritis (EVA). Virus was isolated from nasal swabs and mononuclear cells collected from both stallions
Copyright 1999 Academic Press.
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Publication Date: 1999-07-16 PubMed ID: 10405372DOI: 10.1006/viro.1999.9817Google Scholar: Lookup
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  • 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.

The research is about a form of equine arteritis virus (EAV) that has been genetically modified and is both attenuated and stable. When tested on stallions, the modified virus did not cause any visible signs of illness and remained genetically identical to the original virus.

Study Design and Implementation

  • The study began with the creation of an equine arteritis virus (EAV030H) using a procedure designed to produce an infectious, but weakened (attenuated) virus based on a stable cDNA clone.
  • Two stallions were intranasally inoculated with the genetically engineered virus. The health of the stallions was closely monitored after inoculation.
  • Virus samples were taken from the stallions from their nasal swabs, semen, and mononuclear cells (a type of white blood cell).

Observations and Findings

  • The researchers observed that the horses did not exhibit the symptoms associated with equine viral arteritis (EVA), indicating that the modified virus was indeed attenuated as intended.
  • Isolation and analysis of the virus from different sources (nasal swabs, mononuclear cells, and semen) revealed the virus remained genetically stable.
  • Both horses developed immunity (seroconverted) against EAV and maintained high neutralizing antibody levels. The constant presence of these put the body in a state of readiness to neutralize any active virus entering the body.

Analysis and Interpretation

  • Next, viral RNA was detected using Reverse Transcriptase (RT) nested-PCR, helping to confirm the continued presence of the modified virus without causing disease symptoms.
  • The sequence and restriction digestion analysis established the genetic stability of the virus. The virus found in the nasal swabs, mononuclear cells, and semen was identical to the original virus used for inoculation.
  • The researchers concluded the modified virus stable during replication, a critical finding for potential future applications.

Implications of the research

  • This study reports the first instance where a recombinant virus, derived from an order Nidovirales infectious cDNA, displayed competence to replicate in animals without causing illness.
  • The researchers believed that these findings are significant for the development of new methods to study EAV, particularly the genetic factors which determine virulence (severity) and persistence.

Cite This Article

APA
Balasuriya UB, Snijder EJ, van Dinten LC, Heidner HW, Wilson WD, Hedges JF, Hullinger PJ, MacLachlan NJ. (1999). Equine arteritis virus derived from an infectious cDNA clone is attenuated and genetically stable in infected stallions. Virology, 260(1), 201-208. https://doi.org/10.1006/viro.1999.9817

Publication

Virology
ISSN: 0042-6822
NlmUniqueID: 0110674
Country: United States
Language: English
Volume: 260
Issue: 1
Pages: 201-208

Researcher Affiliations

Balasuriya, U B
  • School of Veterinary Medicine, University of California, One Shields Avenue, Davis, California, 95616, USA.
Snijder, E J
    van Dinten, L C
      Heidner, H W
        Wilson, W D
          Hedges, J F
            Hullinger, P J
              MacLachlan, N J

                MeSH Terms

                • Animals
                • Arterivirus Infections / prevention & control
                • Arterivirus Infections / veterinary
                • Cell Line
                • Cloning, Molecular
                • Cricetinae
                • DNA, Complementary / genetics
                • Equartevirus / genetics
                • Equartevirus / immunology
                • Equartevirus / pathogenicity
                • Horse Diseases / prevention & control
                • Horses
                • Male
                • Polymerase Chain Reaction
                • Rabbits
                • Vaccines, Attenuated
                • Vaccines, DNA

                Citations

                This article has been cited 21 times.
                1. 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).
                  doi: 10.3390/v18010113pubmed: 41600875google scholar: lookup
                2. 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.
                  doi: 10.3389/fgene.2022.871875pubmed: 35495124google scholar: lookup
                3. Câmara RJF, Bueno BL, Resende CF, Balasuriya UBR, Sakamoto SM, Reis JKPD. Viral Diseases that Affect Donkeys and Mules. Animals (Basel) 2020 Nov 25;10(12).
                  doi: 10.3390/ani10122203pubmed: 33255568google scholar: lookup
                4. 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.
                  doi: 10.1371/journal.ppat.1007950pubmed: 31356622google scholar: lookup
                5. 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).
                  doi: 10.1128/JVI.00045-19pubmed: 30918077google scholar: lookup
                6. 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).
                  doi: 10.1128/JVI.00015-18pubmed: 29444949google scholar: lookup
                7. 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).
                  doi: 10.1128/CVI.00215-17pubmed: 28814389google scholar: lookup
                8. 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).
                  doi: 10.1128/JVI.00418-17pubmed: 28424285google scholar: lookup
                9. 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.
                  doi: 10.1016/j.virol.2014.04.029pubmed: 24913633google scholar: lookup
                10. Balasuriya UB, Go YY, MacLachlan NJ. Equine arteritis virus. Vet Microbiol 2013 Nov 29;167(1-2):93-122.
                  doi: 10.1016/j.vetmic.2013.06.015pubmed: 23891306google scholar: lookup
                11. 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.
                  doi: 10.1128/CVI.00302-12pubmed: 22739697google scholar: lookup
                12. 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.
                  doi: 10.1099/vir.0.029264-0pubmed: 21307223google scholar: lookup
                13. 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.
                  doi: 10.1128/CVI.00444-10pubmed: 21147938google scholar: lookup
                14. 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.
                  doi: 10.1128/JVI.02743-09pubmed: 20219931google scholar: lookup
                15. Lu Z, Chambers TM, Boliar S, Branscum AJ, Sturgill TL, Timoney PJ, Reedy SE, Tudor LR, Dubovi EJ, Vickers ML, Sells S, Balasuriya UB. Development and evaluation of one-step TaqMan real-time reverse transcription-PCR assays targeting nucleoprotein, matrix, and hemagglutinin genes of equine influenza virus. J Clin Microbiol 2009 Dec;47(12):3907-13.
                  doi: 10.1128/JCM.00598-09pubmed: 19846644google scholar: lookup
                16. 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.
                  doi: 10.1128/JVI.01249-08pubmed: 18579588google scholar: lookup
                17. 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.
                  doi: 10.1128/CVI.00388-07pubmed: 18032597google scholar: lookup
                18. Yoo D, Welch SK, Lee C, Calvert JG. Infectious cDNA clones of porcine reproductive and respiratory syndrome virus and their potential as vaccine vectors. Vet Immunol Immunopathol 2004 Dec 8;102(3):143-54.
                  doi: 10.1016/j.vetimm.2004.09.019pubmed: 15507301google scholar: lookup
                19. Castillo-Olivares J, Wieringa R, Bakonyi T, de Vries AA, Davis-Poynter NJ, Rottier PJ. Generation of a candidate live marker vaccine for equine arteritis virus by deletion of the major virus neutralization domain. J Virol 2003 Aug;77(15):8470-80.
                  doi: 10.1128/jvi.77.15.8470-8480.2003pubmed: 12857916google scholar: lookup
                20. Bonneau KR, Mullens BA, MacLachlan NJ. Occurrence of genetic drift and founder effect during quasispecies evolution of the VP2 and NS3/NS3A genes of bluetongue virus upon passage between sheep, cattle, and Culicoides sonorensis. J Virol 2001 Sep;75(17):8298-305.
                  doi: 10.1128/jvi.75.17.8298-8305.2001pubmed: 11483775google scholar: lookup
                21. Balasuriya UB, Heidner HW, Hedges JF, Williams JC, Davis NL, Johnston RE, MacLachlan NJ. Expression of the two major envelope proteins of equine arteritis virus as a heterodimer is necessary for induction of neutralizing antibodies in mice immunized with recombinant Venezuelan equine encephalitis virus replicon particles. J Virol 2000 Nov;74(22):10623-30.
                  doi: 10.1128/jvi.74.22.10623-10630.2000pubmed: 11044106google scholar: lookup
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