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Home / Equine Research Bank / Clin Diagn Lab Immunol / Expression cloning and humoral immune response to the nucleo...
Clinical and diagnostic laboratory immunology1998; 4(6); 648-652; doi: 10.1128/cdli.4.6.648-652.1997

Expression cloning and humoral immune response to the nucleocapsid and membrane proteins of equine arteritis virus.

Abstract: To provide a convenient and sensitive method for the detection of equine arteritis virus (EAV)-specific serum antibodies, we developed an immunoblot assay employing the EAV nucleocapsid (N) and membrane (M) proteins expressed in a procaryotic expression vector (pMAL-c2) for the production of recombinant maltose-binding (MBP) fusion proteins (MBP-N and MBP-M). The antigenic reactivity of the recombinant fusion proteins and their Xa factor cleavage EAV products was confirmed by immunoblot using horse antisera to EAV. Some horse sera, however, showed immune reactivity to the MBP fusion partner protein. Based on a total of 32 horse sera analyzed for the presence of EAV antibodies by immunoblot, using the MBP-N or -M fusion proteins and the Xa factor cleavage EAV products, and in the serum neutralization test, there was 100% concordance between the assays. Sera from horses experimentally infected with EAV were reactive in the immunoblot test with both the MBP-N and the MBP-M fusion proteins by day 14 after EAV exposure. The reactivity continued to the end of the experiment at day 145 after infection. This immune reactivity correlated with the detection of neutralizing antibodies in the serum samples. Based on these findings, the recombinant N and M proteins might be useful for serodetection of EAV-infected animals.
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Publication Date: 1998-01-10 PubMed ID: 9384283PubMed Central: PMC170634DOI: 10.1128/cdli.4.6.648-652.1997Google Scholar: Lookup
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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 paper presents a method for detection of antibodies specific to equine arteritis virus (EAV), using recombinant proteins from the virus. The findings show that these proteins can accurately mirror the virus’s presence in the blood serum of horses, thus making them potentially useful in detecting EAV infections.

Method for Detection of EAV-specific Antibodies

  • The researchers developed an immunoblot assay.
  • They used the nucleocapsid (N) and membrane (M) proteins of the EAV, which they expressed in a bacterial expression vector (pMAL-c2).
  • This vector was used to produce recombinant maltose-binding (MBP) fusion proteins.
  • The recombinant proteins (MBP-N and MBP-M) were paired with Xa factor cleavage EAV products to confirm their antigenic reactivity.
  • The antigenic reactivity was confirmed by performing an immunoblot test with horse antisera to EAV.

Findings

  • Some horse sera showed an immune response to the MBP fusion partner protein.
  • Out of 32 horse sera samples tested for EAV antibodies using the recombinant proteins & Xa factor cleavage EAV products, there was a 100% correspondence between the results of the immunoblot test and the serum neutralization test.
  • Samples obtained from horses that were experimentally infected with EAV reacted in the immunoblot test with both the recombinant proteins by day 14 post EAV exposure. This immune reaction persisted till the end of the study, 145 days post-infection.

Implications

  • The immune response observed correlated with the presence of neutralizing antibodies against the virus in the serum samples.
  • This suggests the recombinant N and M proteins could be useful as diagnostic markers for identifying EAV infections in horses.

Cite This Article

APA
Kheyar A, Martin S, St-Laurent G, Timoney PJ, McCollum WH, Archambault D. (1998). Expression cloning and humoral immune response to the nucleocapsid and membrane proteins of equine arteritis virus. Clin Diagn Lab Immunol, 4(6), 648-652. https://doi.org/10.1128/cdli.4.6.648-652.1997

Publication

Clinical and diagnostic laboratory immunology
ISSN: 1071-412X
NlmUniqueID: 9421292
Country: United States
Language: English
Volume: 4
Issue: 6
Pages: 648-652

Researcher Affiliations

Kheyar, A
  • Département des Sciences Biologiques, Université du Québec à Montréal, Canada.
Martin, S
    St-Laurent, G
      Timoney, P J
        McCollum, W H
          Archambault, D

            MeSH Terms

            • ATP-Binding Cassette Transporters
            • Animals
            • Antibodies, Viral / blood
            • Antibody Formation / drug effects
            • Arterivirus Infections / veterinary
            • Blotting, Western
            • Carrier Proteins / biosynthesis
            • Carrier Proteins / genetics
            • Cloning, Molecular
            • Equartevirus / genetics
            • Equartevirus / immunology
            • Escherichia coli / genetics
            • Escherichia coli / metabolism
            • Escherichia coli Proteins
            • Gene Expression
            • Horse Diseases / blood
            • Horse Diseases / immunology
            • Horses
            • Kinetics
            • Maltose-Binding Proteins
            • Monosaccharide Transport Proteins
            • Neutralization Tests
            • Nucleocapsid Proteins / biosynthesis
            • Nucleocapsid Proteins / genetics
            • Nucleocapsid Proteins / pharmacology
            • Open Reading Frames
            • Viral Fusion Proteins / biosynthesis
            • Viral Fusion Proteins / genetics
            • Viral Matrix Proteins / biosynthesis
            • Viral Matrix Proteins / genetics
            • Viral Matrix Proteins / pharmacology

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            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.
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            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.
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            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. Archambault D, Kheyar A, de Vries AA, Rottier PJ. The intraleader AUG nucleotide sequence context is important for equine arteritis virus replication. Virus Genes 2006 Aug;33(1):59-68.
              doi: 10.1007/s11262-005-0030-zpubmed: 16791420google scholar: lookup
            5. 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
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              pubmed: 11768130
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              doi: 10.1128/JCM.38.6.2065-2075.2000pubmed: 10834955google scholar: lookup
            8. Abed Y, St-Laurent G, Zhang H, Jacobs RM, Archambault D. Development of a Western blot assay for detection of bovine immunodeficiency-like virus using capsid and transmembrane envelope proteins expressed from recombinant baculovirus. Clin Diagn Lab Immunol 1999 Mar;6(2):168-72.
              doi: 10.1128/CDLI.6.2.168-172.1999pubmed: 10066648google scholar: lookup
            9. Kheyar A, St-Laurent G, Diouri M, Archambault D. Nucleotide sequence and genetic analysis of the leader region of Canadian, American and European equine arteritis virus isolates. Can J Vet Res 1998 Jul;62(3):224-30.
              pubmed: 9684053
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