FREE SHIPPING over $40
OVER 10000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Virus research1995; 39(2-3); 277-288; doi: 10.1016/0168-1702(95)00098-4

Expression cloning and antigenic analysis of the nucleocapsid protein of equine arteritis virus.

Abstract: A series of recombinant fusion proteins derived from equine arteritis virus (EAV) open reading frame (ORF) 7 have been used to define the immunoreactive region of the viral nucleocapsid (N) protein. Reactivities of recombinant N fusion proteins with post-infection equine sera in immunoblots and ELISAs indicate that the major nucleocapsid protein epitope is located within amino acid residues 1-69. In ELISAs two recombinant nucleocapsid fusion proteins containing residues 1-69 (rN1-69) and 1-28 (rN1-28) discriminated between pre- and post-infection, and pre- and post-vaccination serum samples. Additionally rN1-69 and rN1-28 detected seroconversions following vaccination with a killed virus preparation, even in the absence of a detectable virus neutralising response. Although a good correlation existed between virus neutralising antibody and rN1-69 ELISA positive values in post-infection sera, all the rN proteins failed to induce any virus neutralising response in immunised rabbits.
Get Full Text
Publication Date: 1995-12-01 PubMed ID: 8837890PubMed Central: PMC7133929DOI: 10.1016/0168-1702(95)00098-4Google 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.
LinkedInCopy
  • Journal Article
  • Research Support
  • Non-U.S. Gov't

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 focuses on identifying the main immune-recognized region of the nucleocapsid protein of the equine arteritis virus (EAV) through the use of recombinant fusion proteins. The study finds that this region is located within the first 69 amino acid residues of the protein and is able to differentiate between pre- and post-infection as well as vaccination serum samples.

Concept of the Research

  • The study is centered on the equine arteritis virus (EAV), a virus that affects horses. The researchers specifically look at the nucleocapsid (N) protein of the virus. The N protein is a viral protein associated with the genetic material of the virus.
  • The study uses recombinant fusion proteins, which are proteins created through genetic engineering, to determine important aspects of the N protein. In this case, the recombinant proteins are derived from EAV’s open reading frame (ORF) 7, a part of the virus’ genetic material that can be translated into proteins.

Findings of the Research

  • The researchers discovered that the main epitope, or part of the antigen that is recognized by the immune system, of the N protein is located within the first 69 amino acid residues. Amino acids are the building blocks of proteins and an “amino acid residue” refers to a single amino acid in a peptide chain.
  • In enzyme-linked immunosorbent assays (ELISAs), two recombinant N fusion proteins containing the first 69 (rN1-69) and 28 (rN1-28) residues could differentiate between pre- and post-infection and pre- and post-vaccination serum samples. This suggests these proteins could potentially be used as diagnostic tools.
  • The rN1-69 and rN1-28 proteins could also detect a change in the immune response (seroconversion) following vaccination, even if there was no detectable virus-neutralising response.

Significance of the Research

  • Despite successfully identifying the major nucleocapsid protein epitope, the researchers found that all the recombinant N proteins used in the study failed to induce a virus-neutralizing response in vaccinated rabbits. A virus-neutralizing response refers to the immune system’s ability to inhibit virus infectivity.
  • This suggests that while these proteins could potentially serve as useful diagnostic tools, more research is needed to understand how they could be used in a therapeutic or preventative capacity against EAV.

Cite This Article

APA
Chirnside ED, Francis PM, Mumford JA. (1995). Expression cloning and antigenic analysis of the nucleocapsid protein of equine arteritis virus. Virus Res, 39(2-3), 277-288. https://doi.org/10.1016/0168-1702(95)00098-4

Publication

Virus research
ISSN: 0168-1702
NlmUniqueID: 8410979
Country: Netherlands
Language: English
Volume: 39
Issue: 2-3
Pages: 277-288

Researcher Affiliations

Chirnside, E D
  • Centre for Preventive Medicine, Animal Health Trust, Newmarket, Suffolk, UK.
Francis, P M
    Mumford, J A

      MeSH Terms

      • Animals
      • Antigens, Viral / genetics
      • Antigens, Viral / immunology
      • Base Sequence
      • Cloning, Molecular
      • DNA Primers
      • Equartevirus / genetics
      • Equartevirus / immunology
      • Gene Expression Regulation, Viral
      • Horses
      • Immune Sera / immunology
      • Molecular Sequence Data
      • Nucleocapsid / genetics
      • Nucleocapsid / immunology
      • Rabbits
      • Recombinant Fusion Proteins / genetics
      • Recombinant Fusion Proteins / immunology

      References

      This article includes 33 references
      1. Balasuriya U.B.R., Rossitto P.V., DeMaula C.D., MacLachlan N.J.. A 29K envelope glycoprotein of equine arteriyis virus expresses neutralisation determinants recognised by murine monoclonal antibodies.. J. Gen. Virol. 1993;74:2525–2529.
        pubmed: 7504076
      2. Balasuriya U.B.R., MacLachlan N.J., de Vries A.A.F., Rossitto P.V., Rottier P.J.M.. Identification of a neutralization site in the major envelope glycoprotein (GL) of equine arteritis virus.. Virology 1995;207:518–527.
        pubmed: 7533965
      3. Cavanagh D., Brien D.A., Brinton M., Enjuanes L., Holmes K.V., Horzinek M.C., Lai M.M.C., Laude H., Plagemann P.G.W., Siddell S.. Revision of the taxonomy of the Coronavirus, Torovirus and Arterivirus genera.. Arch. Virol. 1994;135:227–237.
        pmc: PMC7086850pubmed: 8198447
      4. Chirnside E.D.. Equine arteritis virus: an overview.. Br. Vet. J. 1992;148:181–197.
        pmc: PMC7130166pubmed: 1319787
      5. Chirnside E.D., Wearing C.M., Binns M.M., Mumford J.A.. Comparison of M and N gene sequences distinguishes variation amongst equine arteritis virus isolates.. J. Gen. Virol. 1994;74:1491–1497.
        pubmed: 8207415
      6. Chirnside E.D., de Vries A.A.F., Mumford J.A., Rottier P.J.M.. Equine arteritis virus neutralising antibody in the horse is induced by a determinant on the large envelope glycoprotein (GL). J. Gen. Virol. 1995;76:1989–1998.
        pubmed: 7636479
      7. Chirnside E.D., Franvis P.M., de Vries A.A.F., Sinclair R., Mumford J.A.. Development and evaluation of an ELISA using recombinant fusion protein to detect the presence of host antibody to equine arteritis virus (EAV). J. Virol. Methods. 1995;54:1–13.
        pmc: PMC7119792pubmed: 7559853
      8. Conzelmann K.-K., Visser N., van Woensel P., Thiel H.-J.. Molecular characterisation of porcine reproductive and respiratory syndrome virus, a member of the arterivirus group.. Virology 1993;193:329–339.
        pmc: PMC7131490pubmed: 8438574
      9. De Vries A.A.F.. The molecular biology of equine arteritis virus.. The University of Utrecht; 1994. (PhD thesis).
      10. De Vries A.A.F., Chirnside E.D., Bredenbeek P., Gravestein L.A., Horzinek M.C., Spaan W.J.M.. All subgenomic mRNAs of equine arteritis virus contain a common leader sequence.. Nucl. Acids Res. 1990;18:3241–3247.
        pmc: PMC330929pubmed: 2162519
      11. De Vries A.A.F., Chirnside E.D., Horzinek M.C., Rottier P.J.M.. The structural proteins of equine arteritis virus.. J. Virol. 1992;66:6294–6303.
        pmc: PMC240121pubmed: 1328669
      12. De Vries A.A.F., Raamsman M.J.B., van Dijk H.A., Horzinek M.C., Rottier P.J.M.. The small envelope glycoprotein (Gs) of equine arteritis virus folds into three distinct monomers and a disulfide-linked dimer.. J. Virol. 1995;69:3441–3448.
        pmc: PMC189056pubmed: 7745690
      13. Den Boon J.A., Snijder E.J., Chirnside E.D., de Vries A.A.F., Horzinek M.C., Spaan W.J.M.. Equine arteritis virus is not a togavirus but belongs to the coronavirus-like superfamily.. J. Virol. 1991;65:2910–2920.
        pmc: PMC240924pubmed: 1851863
      14. Deregt D., de Vries A.A.F., Raamsman M.J.B., Elmgren L.D., Rottier P.J.M.. Monoclonal antibodies to equine arteritis virus proteins identity the GL protein as a target for virus neutralisation.. J. Gen. Virol. 1994;75:2439–2444.
        pubmed: 8077945
      15. Doll E.R., Knappenberger R.E., Bryans J.T.. An outbreak of abortion caused by the equine arteritis virus.. Cornell Vet. 1957;47:69–75.
        pubmed: 13397180
      16. Gibson T.J.. Studies on the Epstein-Barr virus genome.. University of Cambridge; England: 1984. (PhD thesis).
      17. Godeny E.K., Chen L., Kumar N., Methven S.L., Koonin E.V., Brinton M.A.. Complete genomic sequence and phylogenetic analysis of the lactate dehydrogenase-elevating virus (LDV). Virology 1993;194:585–596.
        pmc: PMC7173116pubmed: 8389075
      18. Golnik W., Michalska Z., Michalak T.. Natural equine viral arteritis in foals.. Schweiz. Arch. Tierheilkd. 1981;123:523–533.
        pubmed: 6274007
      19. Hyllseth B.. Structural proteins of equine arteritis virus.. Arch. gesamte Virusforsch. 1973;40:177–188.
        pubmed: 4633581
      20. Iwashita O., Harasawa R.. Structural polypeptides of equine arteritis virus.. Jpn. J. Vet. Sci. 1987;49:923–925.
        pubmed: 2824911
      21. Kuo L., Chen Z., Rowland R.R.R., Faaberg K.S., Plagemann P.G.W.. Lactate dehydrogenase-elevating virus (LDV): subgenomic mRNAs, mRNA leader and comparison of 3′-terminal sequences of two LDV isolates.. Virus Res. 1992;23:55–72.
        pmc: PMC7133880pubmed: 1604932
      22. McCollum W.H., Doll E.R., Wilson J.C.. The recovery of virus from horses with experimental cases of equine arteritis using monolayer cell cultures of equine kidney.. Am. J. Vet. Res. 1961;23:465–469.
      23. Meulenberg J.J.M., Hulst M.M., de Meijer E.J., Moonen P.L.J.M., van Besten A., de Kluyver E.P., Wensvoort G., Moorman R.J.M.. Lelystad virus, the causative agent of porcine epidemic abortion and respiratory syndrome (PEARS), is related to LDV and EAV.. Virology 1993;192:62–72.
        pmc: PMC7173055pubmed: 8517032
      24. Meulenberg J.J.M., de Meijer E.J., Moorman R.J.M.. Subgenomic RNAs of Leystad virus contain a conserved leader-body junction sequence.. J. Gen. Virol. 1993;74:1697–1701.
        pubmed: 8345361
      25. Murphy F.A.. Togavirus morphology and morphogenesis.. In: Schlesinger R.W., editor. The Togaviruses: Biology Structure and Replication. Academic Press; London: 1980. pp. 241–316.
      26. Plagemann P.G.W., Moennig V.. Lactate dehydrogenase-elevating virus, equine arteritis virus and simian hemorrhagic fever virus: a new group of positive-strand RNA viruses.. Adv. Virus Res. 1992;41:99–192.
        pmc: PMC7131515pubmed: 1315480
      27. Sambrook J., Fritsch E.F., Maniatis T.. Molecular Cloning: A Laboratory Manual. 2nd ed. Cold Spring Harbor Laboratory; New York: 1989..
      28. Senne D.A., Pearson J.E., Carbrey E.A.. Equine viral arteritis: a standard procedure for the virus neutralisation test and comparison of results of a proficiency test performed at five laboratories.. Proceedings of the 89th Annual Meeting of the United States Animal Health Association Milwaukee, WI; 1985. pp. 29–34.
      29. Smith D.B., Johnson K.S.. Single-step purification of polypeptides expressed in Escherichia coli as fusion proteins with glutathione-S-transferase.. Gene 1988;67:31–40.
        pubmed: 3047011
      30. Timoney P.J., McCollum W.H., Roberts A.W., Murphy T.W.. Demonstration of the carrier state in naturally acquired equine arteritis virus infection in the stallion.. Res. Vet. Sci. 1986;41:279–280.
        pubmed: 3022363
      31. Vaala W.E., Hamir A.N., Dubovi E.J., Timoney P.J., Ruiz B.. Fatal, congenitally acquired infection with equine arteritis virus in a neonatal thoroughbred.. Equine Vet. J. 1992;24:155–158.
        pubmed: 1316264
      32. Van Berlo M.F., Rottier P.J.M., Spaan W.J.M., Horzinek M.C.. Equine arteritis virus (EAV) induced polypeptide synthesis.. J. Gen. Virol. 1986;67:1543–1549.
        pubmed: 2426393
      33. Zeegers J.J.W., van der Zeijst B.A.M., Horzinek M.C.. The structural proteins of equine arteritis virus.. Virology 1976;73:200–205.
        pubmed: 183352

      Citations

      This article has been cited 9 times.
      1. 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
      2. Zhao S, Qi T, Guo W, Lu G, Xiang W. Identification of a conserved B-cell epitope in the equine arteritis virus (EAV) N protein using the pepscan technique. Virus Genes 2013 Oct;47(2):292-7.
        doi: 10.1007/s11262-013-0943-xpubmed: 23813249google scholar: lookup
      3. 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
      4. 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
      5. Glaser AL, Chirnside ED, Horzinek MC, de Vries AA. Equine arteritis virus. Theriogenology 1997 Apr 15;47(6):1275-95.
        doi: 10.1016/s0093-691x(97)00107-6pubmed: 16728076google scholar: lookup
      6. Smits SL, Snijder EJ, de Groot RJ. Characterization of a torovirus main proteinase. J Virol 2006 Apr;80(8):4157-67.
        doi: 10.1128/JVI.80.8.4157-4167.2006pubmed: 16571831google scholar: lookup
      7. Jeronimo C, Archambault D. Importance of M-protein C terminus as substrate antigen for serodetection of equine arteritis virus infection. Clin Diagn Lab Immunol 2002 May;9(3):698-703.
        doi: 10.1128/cdli.9.3.698-703.2002pubmed: 11986280google scholar: lookup
      8. 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.
        doi: 10.1128/JCM.38.6.2065-2075.2000pubmed: 10834955google scholar: lookup
      9. Kheyar A, Martin S, St-Laurent G, Timoney PJ, McCollum WH, Archambault D. Expression cloning and humoral immune response to the nucleocapsid and membrane proteins of equine arteritis virus. Clin Diagn Lab Immunol 1997 Nov;4(6):648-52.
        doi: 10.1128/cdli.4.6.648-652.1997pubmed: 9384283google scholar: lookup
      Subscribe to our newsletter
      Join 99,911 horse owners like you who receive our email updates on horse health and nutrition.