Serologic response of horses to the structural proteins of equine arteritis virus.
Abstract: Equine arteritis virus (EAV) is the causative agent of equine viral arteritis, an apparently emerging disease of equids. In this study, the antibody response of horses to the structural proteins of EAV was evaluated using gradient-purified EAV virions and baculovirus-expressed recombinant EAV structural proteins (G(L), G(S), M, N) as antigens in a Western immunoblotting assay. Thirty-three sera from horses that previously had been naturally or experimentally infected with EAV were evaluated, including samples from mares, geldings, and both persistently and nonpersistently infected stallions. Sera also were evaluated from 4 horses that had been vaccinated with the commercial modified live EAV vaccine. The data suggest that the serologic response of individual horses to EAV may vary with the infecting virus strain and duration of infection. The M protein was most consistently recognized by the various serum samples, whereas the response to the N and G(L) proteins was variable and the G(S) protein was bound by only 1 serum sample. The immunoblotting assay definitively established the protein specificity of the humoral response of horses to EAV; however, it clearly is less sensitive than the standard serum neutralization (SN) test--2 of the 37 sera that were seropositive by the SN test failed to react in the immunoblot assay with any EAV structural protein. Furthermore, the G(L) protein expresses the known neutralization determinants of EAV, yet only 22 of the 37 sera that had SN antibodies bound the G(L) protein in the immunoblotting assay. Information from this study will assist ongoing efforts to develop improved methods for the serologic diagnosis of EAV infection of horses.
Publication Date: 1998-07-31 PubMed ID: 9683071DOI: 10.1177/104063879801000302Google 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
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The research investigates the structural proteins of Equine Arteritis Virus (EAV), identifying three virus-specific proteins, and adds to the evidence suggesting EAV’s inclusion in the Togaviridae family.
Research Method
- The research team carried out the study using Equine arteritis virus (EAV) cultured in three different kinds of cells – Vero, BHK-21, and RK-13.
- The virus was then purified through a three-step process of pelleting, Sepharose 6B chromatography, and sucrose gradient centrifugation.
- Afterwards, the entire gradients were analyzed by means of polyacrylamide slab gel electrophoresis followed by autoradiography. This revealed a substantial number of proteins in all the fractions.
Findings
- The researchers noted differences between the protein patterns in the virus when grown in the separate cell types. However, three specific proteins appeared to be unique to EAV itself. These were labelled VP1, VP2, and VP3, with molecular weights of 12,000, 14,000, and 21,000 respectively.
- The VP1 protein is described as a phosphorylated core protein. In the world of cell biology, phosphorylation is a process crucial to many cell functions, like cell division and growth.
- VP3, on the other hand, is identified as a glycoprotein, which is a protein that has carbohydrate chains attached to it. Glycoproteins play critical roles in immune response and in functioning of cells.
Implications
- These results, along with previous findings regarding the virus’s morphology and RNA, provide further evidence supporting the placement of EAV in the Togaviridae virus family.
- This underscores a potential correlation to lactic dehydrogenase virus, another member of the Togaviridae family. Lactic dehydrogenase virus is a significant infectious agent in mice, capable of causing serious pathogenic effects.
- If this relation is confirmed, it could provide valuable insights into understanding the EAV and developing effective treatments.
Cite This Article
APA
MacLachlan NJ, Balasuriya UB, Hedges JF, Schweidler TM, McCollum WH, Timoney PJ, Hullinger PJ, Patton JF.
(1998).
Serologic response of horses to the structural proteins of equine arteritis virus.
J Vet Diagn Invest, 10(3), 229-236.
https://doi.org/10.1177/104063879801000302 Publication
Researcher Affiliations
- Department of Pathology, Microbiology, and Immunology, School of Veterinary Medicine, University of California, Davis 95616, USA.
MeSH Terms
- Animals
- Antibodies, Viral / blood
- Antibody Formation
- Antigens, Viral / genetics
- Antigens, Viral / immunology
- Arterivirus Infections / immunology
- Arterivirus Infections / prevention & control
- Arterivirus Infections / veterinary
- DNA Primers
- Equartevirus / genetics
- Equartevirus / immunology
- Horse Diseases / immunology
- Horse Diseases / prevention & control
- Horses
- Open Reading Frames
- Polymerase Chain Reaction
- Recombinant Proteins / immunology
- Viral Structural Proteins / genetics
- Viral Structural Proteins / immunology
- Viral Vaccines
- Virion / immunology
Citations
This article has been cited 21 times.- Mayers J, Westcott D, Steinbach F. Identification of Equine Arteritis Virus Immunodominant Epitopes Using a Peptide Microarray. Viruses 2022 Aug 26;14(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).
- Oudshoorn D, van der Hoeven B, Limpens RW, Beugeling C, Snijder EJ, Bárcena M, Kikkert M. Antiviral Innate Immune Response Interferes with the Formation of Replication-Associated Membrane Structures Induced by a Positive-Strand RNA Virus. mBio 2016 Dec 6;7(6).
- Balasuriya UB, Go YY, MacLachlan NJ. Equine arteritis virus. Vet Microbiol 2013 Nov 29;167(1-2):93-122.
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- 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.
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- 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.
- Zhang J, Stein DA, Timoney PJ, Balasuriya UB. Curing of HeLa cells persistently infected with equine arteritis virus by a peptide-conjugated morpholino oligomer. Virus Res 2010 Jun;150(1-2):138-42.
- Nedialkova DD, Gorbalenya AE, Snijder EJ. Arterivirus Nsp1 modulates the accumulation of minus-strand templates to control the relative abundance of viral mRNAs. PLoS Pathog 2010 Feb 19;6(2):e1000772.
- 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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- Tijms MA, van Dinten LC, Gorbalenya AE, Snijder EJ. A zinc finger-containing papain-like protease couples subgenomic mRNA synthesis to genome translation in a positive-stranded RNA virus. Proc Natl Acad Sci U S A 2001 Feb 13;98(4):1889-94.
- Pasternak AO, Gultyaev AP, Spaan WJ, Snijder EJ. Genetic manipulation of arterivirus alternative mRNA leader-body junction sites reveals tight regulation of structural protein expression. J Virol 2000 Dec;74(24):11642-53.
- 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.
- Weiland E, Bolz S, Weiland F, Herbst W, Raamsman MJ, Rottier PJ, De Vries AA. Monoclonal antibodies directed against conserved epitopes on the nucleocapsid protein and the major envelope glycoprotein of equine arteritis virus. J Clin Microbiol 2000 Jun;38(6):2065-75.
- van Dinten LC, van Tol H, Gorbalenya AE, Snijder EJ. The predicted metal-binding region of the arterivirus helicase protein is involved in subgenomic mRNA synthesis, genome replication, and virion biogenesis. J Virol 2000 Jun;74(11):5213-23.
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