FREE SHIPPING over $40
OVER 10000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
The Journal of general virology1995; 76 ( Pt 9); 2223-2233; doi: 10.1099/0022-1317-76-9-2223

Comparison of equine arteritis virus isolates using neutralizing monoclonal antibodies and identification of sequence changes in GL associated with neutralization resistance.

Abstract: Three murine monoclonal antibodies (MAbs) that neutralize equine arteritis virus (EAV) infectivity were identified and characterized. The antibodies, 93B, 74D(B) and 38F, recognized the major envelope glycoprotein (GL) encoded by open reading frame (ORF) 5 in immunoblots and by immunoprecipitation. All three MAbs were used to compare the Bucyrus isolate of EAV and MAb neutralization-resistant (NR) escape mutants with the vaccine virus and 19 independent field isolates of EAV by virus neutralization. The different abilities of the MAbs to neutralize virus isolates indicated that they recognize non-identical epitopes. Susceptibility to virus neutralization could not be used to distinguish viruses from acutely and persistently infected horses. Comparison of the ORF 5 nucleotide and deduced amino acid sequence from NR and neutralization-sensitive virus isolates revealed amino acid sequence changes at positions 99 and 100 which correlate with the NR phenotype. Additional unique changes in the amino acid sequence of MAb NR viruses at positions 96 and 113 may also contribute to neutralization resistance. The sequence data further showed that the Bucyrus-derived viruses contain one N-glycosylation site, whereas the field isolates DL8 and DL11 possess two sites, both of which are used. Most of the non-conservative amino acid sequence changes were located within the second half of the N-terminal hydrophilic domain. Sequence changes within the first half of the N-terminal ectodomain, the predicted transmembrane domain and the C-terminal hydrophilic domain were mainly silent base substitutions or resulted in conservative amino acid substitutions, suggesting that these regions of the protein are functionally conserved.
Get Full Text
Publication Date: 1995-09-01 PubMed ID: 7561759DOI: 10.1099/0022-1317-76-9-2223Google 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
  • Comparative Study
  • Journal Article

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 discusses the identification and characterization of three murine monoclonal antibodies known for neutralizing equine arteritis virus. It examines the sequence changes that render the virus immune to neutralization and how these changes can’t be used to differentiate viruses from acutely and persistently infected horses.

Identified and Characterized Monoclonal Antibodies

  • The study successfully identified three monoclonal antibodies, named 93B, 74D(B), and 38F, that neutralize the equine arteritis virus (EAV) infectivity.
  • The antibodies were confirmed to recognize the major envelope glycoprotein (GL), which is encoded by open reading frame (ORF) 5. This was established through immunoblots and immunoprecipitation methods.
  • These antibodies were then used to compare various EAV isolates such as the Bucyrus isolate, MAb neutralization-resistant escape mutants, the vaccine virus, and 19 independent field isolates.

Neutralization Resistance and Nucleotide Sequence Changes

  • The research found different capabilities of the antibodies to neutralize the virus isolates, which indicated that the antibodies recognize non-identical epitopes or parts of the virus.
  • Neutralization susceptibility was not found to be effective in distinguishing viruses from acutely and persistively infected horses.
  • Amino acid sequence alterations at positions 99 and 100 were found in neutralization-resistant and neutralization-sensitive virus isolates. These changes correlated with the neutralization-resistant phenotype.
  • Other unique sequence changes at positions 96 and 113 might also contribute to the resistance to neutralization by these antibodies.

Glycosylation Sites and Sequence Changes

  • The Bucyrus-derived virus isolates were found to contain one N-glycosylation site, which is a biochemical process necessary for the functioning of proteins. In contrast, the field isolates DL8 and DL11 contained two such sites, both of which were utilized.
  • Most of the non-conservative amino acid sequence alterations were located within the second half of the N-terminal hydrophilic domain, suggesting that significant changes occur in this region.
  • On the other hand, the sequence changes in the first half of the N-terminal ectodomain, the predicted transmembrane domain, and the C-terminal hydrophilic domain mostly led to silent base substitutions or resulted in conservative amino acid substitutions. This indicates that these regions of the glycoprotein are functionally conserved and maintain their structure and function despite slight changes in the sequence.

Cite This Article

APA
Glaser AL, de Vries AA, Dubovi EJ. (1995). Comparison of equine arteritis virus isolates using neutralizing monoclonal antibodies and identification of sequence changes in GL associated with neutralization resistance. J Gen Virol, 76 ( Pt 9), 2223-2233. https://doi.org/10.1099/0022-1317-76-9-2223

Publication

The Journal of general virology
ISSN: 0022-1317
NlmUniqueID: 0077340
Country: England
Language: English
Volume: 76 ( Pt 9)
Pages: 2223-2233

Researcher Affiliations

Glaser, A L
  • Diagnostic Laboratory, New York State College of Veterinary Medicine, Cornell University, Ithaca 14852-5786, USA.
de Vries, A A
    Dubovi, E J

      MeSH Terms

      • Amino Acid Sequence
      • Animals
      • Antibodies, Monoclonal / immunology
      • Antibodies, Viral / immunology
      • Base Sequence
      • DNA, Viral / analysis
      • Epitopes / immunology
      • Equartevirus / genetics
      • Equartevirus / immunology
      • Equartevirus / isolation & purification
      • Glycoproteins / genetics
      • Glycoproteins / immunology
      • Glycosylation
      • Mice
      • Mice, Inbred BALB C
      • Molecular Sequence Data
      • Neutralization Tests
      • Open Reading Frames
      • RNA, Viral / analysis
      • Sequence Homology, Amino Acid
      • Sequence Homology, Nucleic Acid
      • Viral Envelope Proteins / genetics
      • Viral Envelope Proteins / immunology

      Citations

      This article has been cited 30 times.
      1. Mehdizadeh Gohari I, Brefo-Mensah EK, Palmer M, Boerlin P, Prescott JF. Sialic acid facilitates binding and cytotoxic activity of the pore-forming Clostridium perfringens NetF toxin to host cells. PLoS One 2018;13(11):e0206815.
        doi: 10.1371/journal.pone.0206815pubmed: 30403719google scholar: lookup
      2. 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
      3. Mehdizadeh Gohari I, Parreira VR, Nowell VJ, Nicholson VM, Oliphant K, Prescott JF. A novel pore-forming toxin in type A Clostridium perfringens is associated with both fatal canine hemorrhagic gastroenteritis and fatal foal necrotizing enterocolitis. PLoS One 2015;10(4):e0122684.
        doi: 10.1371/journal.pone.0122684pubmed: 25853427google scholar: lookup
      4. Steinbach F, Westcott DG, McGowan SL, Grierson SS, Frossard JP, Choudhury B. Re-emergence of a genetic outlier strain of equine arteritis virus: Impact on phylogeny. Virus Res 2015 Apr 16;202:144-50.
        doi: 10.1016/j.virusres.2014.12.009pubmed: 25527462google scholar: lookup
      5. 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
      6. 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
      7. 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
      8. 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
      9. 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
      10. 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
      11. Wieringa R, de Vries AA, van der Meulen J, Godeke GJ, Onderwater JJ, van Tol H, Koerten HK, Mommaas AM, Snijder EJ, Rottier PJ. Structural protein requirements in equine arteritis virus assembly. J Virol 2004 Dec;78(23):13019-27.
        doi: 10.1128/JVI.78.23.13019-13027.2004pubmed: 15542653google scholar: lookup
      12. 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
      13. Snijder EJ, Dobbe JC, Spaan WJ. Heterodimerization of the two major envelope proteins is essential for arterivirus infectivity. J Virol 2003 Jan;77(1):97-104.
        doi: 10.1128/jvi.77.1.97-104.2003pubmed: 12477814google scholar: lookup
      14. 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
      15. 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
      16. 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
      17. 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.
        doi: 10.1128/jvi.74.11.5213-5223.2000pubmed: 10799597google scholar: lookup
      18. van Marle G, Dobbe JC, Gultyaev AP, Luytjes W, Spaan WJ, Snijder EJ. Arterivirus discontinuous mRNA transcription is guided by base pairing between sense and antisense transcription-regulating sequences. Proc Natl Acad Sci U S A 1999 Oct 12;96(21):12056-61.
        doi: 10.1073/pnas.96.21.12056pubmed: 10518575google scholar: lookup
      19. Snijder EJ, van Tol H, Pedersen KW, Raamsman MJ, de Vries AA. Identification of a novel structural protein of arteriviruses. J Virol 1999 Aug;73(8):6335-45.
        doi: 10.1128/JVI.73.8.6335-6345.1999pubmed: 10400725google scholar: lookup
      20. van Dinten LC, Rensen S, Gorbalenya AE, Snijder EJ. Proteolytic processing of the open reading frame 1b-encoded part of arterivirus replicase is mediated by nsp4 serine protease and Is essential for virus replication. J Virol 1999 Mar;73(3):2027-37.
        doi: 10.1128/JVI.73.3.2027-2037.1999pubmed: 9971783google scholar: lookup
      21. Pedersen KW, van der Meer Y, Roos N, Snijder EJ. Open reading frame 1a-encoded subunits of the arterivirus replicase induce endoplasmic reticulum-derived double-membrane vesicles which carry the viral replication complex. J Virol 1999 Mar;73(3):2016-26.
        doi: 10.1128/JVI.73.3.2016-2026.1999pubmed: 9971782google scholar: lookup
      22. 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
      23. Pirzadeh B, Gagnon CA, Dea S. Genomic and antigenic variations of porcine reproductive and respiratory syndrome virus major envelope GP5 glycoprotein. Can J Vet Res 1998 Jul;62(3):170-7.
        pubmed: 9684045
      24. van der Meer Y, van Tol H, Locker JK, Snijder EJ. ORF1a-encoded replicase subunits are involved in the membrane association of the arterivirus replication complex. J Virol 1998 Aug;72(8):6689-98.
        doi: 10.1128/JVI.72.8.6689-6698.1998pubmed: 9658116google scholar: lookup
      25. 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
      26. Meulenberg JJ, van Nieuwstadt AP, van Essen-Zandbergen A, Langeveld JP. Posttranslational processing and identification of a neutralization domain of the GP4 protein encoded by ORF4 of Lelystad virus. J Virol 1997 Aug;71(8):6061-7.
        doi: 10.1128/JVI.71.8.6061-6067.1997pubmed: 9223499google scholar: lookup
      27. van Dinten LC, den Boon JA, Wassenaar AL, Spaan WJ, Snijder EJ. An infectious arterivirus cDNA clone: identification of a replicase point mutation that abolishes discontinuous mRNA transcription. Proc Natl Acad Sci U S A 1997 Feb 4;94(3):991-6.
        doi: 10.1073/pnas.94.3.991pubmed: 9023370google scholar: lookup
      28. St-Laurent G, Lepage N, Carman S, Archambault D. Genetic and amino acid analysis of the GL protein of Canadian, American and European equine arteritis virus isolates. Can J Vet Res 1997 Jan;61(1):72-6.
        pubmed: 9008807
      29. Lepage N, St-Laurent G, Carman S, Archambault D. Comparison of nucleic and amino acid sequences and phylogenetic analysis of the Gs protein of various equine arteritis virus isolates. Virus Genes 1996;13(1):87-91.
        doi: 10.1007/BF00576983pubmed: 8938984google scholar: lookup
      30. Hedges JF, Balasuriya UB, Timoney PJ, McCollum WH, MacLachlan NJ. Genetic variation in open reading frame 2 of field isolates and laboratory strains of equine arteritis virus. Virus Res 1996 Jun;42(1-2):41-52.
        doi: 10.1016/0168-1702(96)01294-4pubmed: 8806173google scholar: lookup
      Subscribe to our newsletter
      Join 99,727 horse owners like you who receive our email updates on horse health and nutrition.