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PloS one2024; 19(7); e0301987; doi: 10.1371/journal.pone.0301987

Antibody reactions of horses against various domains of the EHV-1 receptor-binding protein gD1.

Abstract: Equid alphaherpesviruses 1 (EHV-1) and 4 (EHV-4) are closely related and both endemic in horses worldwide. Both viruses replicate in the upper respiratory tract, but EHV-1 may additionally lead to abortion and equine herpesvirus myeloencephalopathy (EHM). We focused on antibody responses in horses against the receptor-binding glycoprotein D of EHV-1 (gD1), which shares a 77% amino acid identity with its counterpart in EHV-4 (gD4). Both antigens give rise to cross-reacting antibodies, including neutralizing antibodies. However, immunity against EHV-4 is not considered protective against EHM. While a diagnostic ELISA to discriminate between EHV-1 and EHV-4 infections is available based on type-specific fragments of glycoprotein G (gG1 and gG4, respectively), the type-specific antibody reaction against gD1 has not yet been sufficiently addressed. Starting from the N-terminus of gD1, we developed luciferase immunoprecipitation system (LIPS) assays, using gD1-fragments of increasing size as antigens, i.e. gD1_83 (comprising the first 83 amino acids), gD1_160, gD1_180, and gD1_402 (the full-length molecule). These assays were then used to analyse panels of horse sera from Switzerland (n = 60) and Iceland (n = 50), the latter of which is considered EHV-1 free. We detected only one true negative horse serum from Iceland, whereas all other sera in both panels were seropositive for both gG4 (ELISA) and gD1 (LIPS against gD1_402). In contrast, seropositivity against gG1 was rather rare (35% Swiss sera; 14% Icelandic sera). Therefore, a high percentage of antibodies against gD1 could be attributed to cross-reaction and due to EHV-4 infections. In contrast, the gD1_83 fragment was able to identify sera with type-specific antibodies against gD1. Interestingly, those sera stemmed almost exclusively from vaccinated horses. Although it is uncertain that the N-terminal epitopes of gD1 addressed in this communication are linked to better protection, we suggest that in future vaccine developments, type-common antigens should be avoided, while a broad range of type-specific antigens should be favored.
Copyright: © 2024 Schramm et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
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Publication Date: 2024-07-12 PubMed ID: 38995916PubMed Central: PMC11244823DOI: 10.1371/journal.pone.0301987Google 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.

The research article focuses on the antibody responses in horses against a specific protein present in Equid alphaherpesviruses-1 (EHV-1). The study uses luciferase immunoprecipitation system (LIPS) assays and also discusses implications of the findings on potential vaccine developments.

Understanding the Research Background

  • Equid alphaherpesviruses 1 (EHV-1) and 4 (EHV-4) are closely related viruses, which cause infections in horses globally. Both these viruses primarily replicate in the upper respiratory tract. However, EHV-1 also results sometimes in abortion and equine herpesvirus myeloencephalopathy (EHM).
  • The study examines the antibody responses elicited in horses against the glycoprotein D of EHV-1 (gD1), which is a protein involved in virus entry into cells.
  • The gD1 protein shares 77% of its amino acid sequence similarity with the equivalent protein found in EHV-4 (gD4). As a result, it is common for both proteins to instigate the production of cross-reacting antibodies, including neutralizing antibodies that can inhibit a virus’s ability to infect cells.
  • Despite an existing immunity against EHV-4, it does not offer protection against the more severe symptoms of EHV-1 infections like EHM.

The Research Methodology and Findings

  • The researchers developed luciferase immunoprecipitation system (LIPS) assays, in which different sizes of the gD1 protein were used as antigens, to understand the antibody reactions against gD1 protein in horses.
  • The LIPS assays were tested on blood serum samples from horses in Switzerland (60 samples) and Iceland (50 samples). Iceland is considered free of EHV-1.
  • All the serum samples, barring one from Iceland, were found to have antibodies against both gD1 and gG4 (another protein in EHV-4), detected through the LIPS and ELISA methods respectively, indicating most horses were previously exposed to an infection with EHV-4.
  • However, only 35% of Swiss samples and 14% of Icelandic samples had antibodies against gG1, implying a high percentage of antibodies against gD1 are likely due to cross-reaction resulting from EHV-4 infections.
  • Interestingly, a smaller fragment of the gD1 protein (gD1_83) was found to be successful in recognizing type-specific antibodies against gD1, and these were mostly found in vaccinated horses.

Implications and Recommendations

  • While the study does not conclusively establish that the presence of antibodies against specific areas (epitopes) of gD1 provides better protection, it suggests that future vaccine developments should focus on type-specific antigens.
  • Researchers recommend avoiding type-common antigens that could lead to cross-reactivity and lack of specific immunity against EHV-1 or EHV-4 infections.

Cite This Article

APA
Schramm A, Ackermann M, Eichwald C, Aguilar C, Fraefel C, Lechmann J. (2024). Antibody reactions of horses against various domains of the EHV-1 receptor-binding protein gD1. PLoS One, 19(7), e0301987. https://doi.org/10.1371/journal.pone.0301987

Publication

PloS one
ISSN: 1932-6203
NlmUniqueID: 101285081
Country: United States
Language: English
Volume: 19
Issue: 7
Pages: e0301987

Researcher Affiliations

Schramm, Andreina
  • Institute of Virology, Vetsuisse Faculty, University of Zurich, Zurich, Switzerland.
Ackermann, Mathias
  • Institute of Virology, Vetsuisse Faculty, University of Zurich, Zurich, Switzerland.
Eichwald, Catherine
  • Institute of Virology, Vetsuisse Faculty, University of Zurich, Zurich, Switzerland.
Aguilar, Claudio
  • Institute of Virology, Vetsuisse Faculty, University of Zurich, Zurich, Switzerland.
Fraefel, Cornel
  • Institute of Virology, Vetsuisse Faculty, University of Zurich, Zurich, Switzerland.
Lechmann, Julia
  • Institute of Virology, Vetsuisse Faculty, University of Zurich, Zurich, Switzerland.

MeSH Terms

  • Animals
  • Horses / immunology
  • Herpesvirus 1, Equid / immunology
  • Antibodies, Viral / immunology
  • Antibodies, Viral / blood
  • Viral Envelope Proteins / immunology
  • Horse Diseases / virology
  • Horse Diseases / immunology
  • Horse Diseases / prevention & control
  • Herpesvirus 4, Equid / immunology
  • Herpesviridae Infections / veterinary
  • Herpesviridae Infections / immunology
  • Herpesviridae Infections / virology
  • Cross Reactions / immunology
  • Enzyme-Linked Immunosorbent Assay
  • Antibodies, Neutralizing / immunology
  • Antibodies, Neutralizing / blood
  • Protein Domains / immunology

Conflict of Interest Statement

The authors have declared that no competing interests exist.

References

This article includes 42 references
  1. Gatherer D, Depledge DP, Hartley CA, Szpara ML, Vaz PK, Benkő M. ICTV virus taxonomy profile: Herpesviridae 2021. Journal of General Virology 2021;102.
    doi: 10.1099/jgv.0.001673pmc: PMC8604186pubmed: 34704922google scholar: lookup
  2. Kydd JH, Townsend HGG, Hannant D. The equine immune response to equine herpesvirus-1: The virus and its vaccines. Vet Immunol Immunopathol 2006;111: 15–30.
    doi: 10.1016/j.vetimm.2006.01.005pubmed: 16476492google scholar: lookup
  3. Thorsteinsdóttir L, Torfason EG, Torsteinsdóttir S, Svansson V. Genetic diversity of equine gammaherpesviruses (γ-EHV) and isolation of a syncytium forming EHV-2 strain from a horse in Iceland. Res Vet Sci 2013;94: 170–177.
    doi: 10.1016/j.rvsc.2012.07.011pubmed: 22862856google scholar: lookup
  4. Torfason EG, Thorsteinsdóttir L, Torsteinsdóttir S, Svansson V. Study of equid herpesviruses 2 and 5 in Iceland with a type-specific polymerase chain reaction. Res Vet Sci 2008;85: 605–611.
    doi: 10.1016/j.rvsc.2008.01.003pubmed: 18336849google scholar: lookup
  5. Thórhallsdóttir K, Svansson V, Larsen LE. Investigations of equine alphaherpesvirus type 1 and 4 infections in a group of foals and their dams in Iceland. Technical University of Denmark 2013.
  6. Ma G, Azab W, Osterrieder N. Equine herpesviruses type 1 (EHV-1) and 4 (EHV-4) —Masters of co-evolution and a constant threat to equids and beyond. Vet Microbiol 2013;167: 123–134.
    doi: 10.1016/j.vetmic.2013.06.018pubmed: 23890672google scholar: lookup
  7. Patel JR, Heldens J. Equine herpesviruses 1 (EHV-1) and 4 (EHV-4)—Epidemiology, disease and immunoprophylaxis: A brief review. Veterinary Journal 2005;170: 14–23.
    doi: 10.1016/j.tvjl.2004.04.018pubmed: 15993786google scholar: lookup
  8. Khusro A, Aarti C, Rivas-Caceres RR, Barbabosa -Pliego A. Equine Herpesvirus-I Infection in Horses: Recent Updates on its Pathogenicity, Vaccination, and Preventive Management Strategies. J Equine Vet Sci 2020;87: 102923.
    doi: 10.1016/j.jevs.2020.102923pubmed: 32172913google scholar: lookup
  9. Osterrieder N, Van de Walle GR. Pathogenic potential of equine alphaherpesviruses: The importance of the mononuclear cell compartment in disease outcome. Vet Microbiol 2010;143: 21–28.
    doi: 10.1016/j.vetmic.2010.02.010pubmed: 20202764google scholar: lookup
  10. SMITH KC, WHITWELL KE, BINNS MM, DOLBY CA, HANNANT D, MUMFORD JA. Abortion of virologically negative foetuses following experimental challenge of pregnant pony mares with Equid herpesvirus 1. Equine Vet J 1992;24: 256–259.
    doi: 10.1111/j.2042-3306.1992.tb02830.xpubmed: 1323457google scholar: lookup
  11. Wilson WD. Equine Herpesvirus 1 Myeloencephalopathy. Veterinary Clinics of North America: Equine Practice 1997;13: 53–72.
    doi: 10.1016/s0749-0739(17)30255-9pubmed: 9106343google scholar: lookup
  12. Lunn DP, Horohov DW, Osterrieder K, Pusterla N, Townsend HGG. EHV-1 Consensus Statement. J Vet Intern Med 2009;23: 450–461.
    pubmed: 19645832
  13. Nugent J, Birch-Machin I, Smith KC, Mumford JA, Swann Z, Newton JR. Analysis of Equid Herpesvirus 1 Strain Variation Reveals a Point Mutation of the DNA Polymerase Strongly Associated with Neuropathogenic versus Nonneuropathogenic Disease Outbreaks. J Virol 2006;80: 4047–4060.
    doi: 10.1128/JVI.80.8.4047-4060.2006pmc: PMC1440451pubmed: 16571821google scholar: lookup
  14. Marenzoni ML, De Waure C, Timoney PJ. Efficacy of vaccination against equine herpesvirus type 1 (EHV-1) infection: Systematic review and meta-analysis of randomised controlled challenge trials. Equine Vet J 2022.
    doi: 10.1111/evj.13870pubmed: 35946376google scholar: lookup
  15. Duvaxyn® EHV 1,4 ad us. vet. Jun 2017 [cited 10 Jul 2023]. Available: https://www.vetpharm.uzh.ch/tak/00000000/00001458.01.
  16. Pusterla N, David Wilson W, Madigan JE, Ferraro GL. Equine herpesvirus-1 myeloencephalopathy: A review of recent developments. Veterinary Journal 2009;180: 279–289.
    doi: 10.1016/j.tvjl.2008.08.004pubmed: 18805030google scholar: lookup
  17. Azab W, Osterrieder N. Glycoproteins D of Equine Herpesvirus Type 1 (EHV-1) and EHV-4 Determine Cellular Tropism Independently of Integrins. J Virol 2012;86: 2031–2044.
    doi: 10.1128/JVI.06555-11pmc: PMC3302398pubmed: 22171258google scholar: lookup
  18. Holz CL, Nelli RK, Eilidh Wilson M, Zarski LM, Azab W, Baumgardner R. Viral genes and cellular markers associated with neurological complications during herpesvirus infections. Journal of General Virology 2017;98: 1439–1454.
    doi: 10.1099/jgv.0.000773pubmed: 28631601google scholar: lookup
  19. Edington N B. One way protection between equid herpesvirus 1 and 4 in vivo. Res Vet Sci 1990;48: 235–239.
    pubmed: 2159176
  20. Telford EAR, Watson MS, Perry J, Cullinane AA, Davison AJ. The DNA sequence of equine herpesvirus-1. Journal of General Virology 1992;79: 1197–1203.
    doi: 10.1016/0042-6822(92)90706-upubmed: 9603335google scholar: lookup
  21. Telford EAR, Watson MS, Perry J, Cullinane AA, Davison AJ. The DNA sequence of equine herpesvirus-4. Journal of General Virology 1998;79: 1197–1203.
    doi: 10.1099/0022-1317-79-5-1197pubmed: 9603335google scholar: lookup
  22. Burbelo PD, Goldman R, Mattson TL. A simplified immunoprecipitation method for quantitatively measuring antibody responses in clinical sera samples by using mammalian-produced Renilla luciferase-antigen fusion proteins. BMC Biotechnol 2005;5.
    doi: 10.1186/1472-6750-5-22pmc: PMC1208859pubmed: 16109166google scholar: lookup
  23. Burbelo PD, Lebovitz EE, Notkins AL. Luciferase immunoprecipitation systems for measuring antibodies in autoimmune and infectious diseases. Translational Research 2015.
    doi: 10.1016/j.trsl.2014.08.006pmc: PMC4306608pubmed: 25241936google scholar: lookup
  24. Li J, Wang G, Yang D, Zhao B, Zhao Y, Liu Y. Development of luciferase-linked antibody capture assay based on luciferase immunoprecipitation systems for antibody detection of porcine reproductive and respiratory syndrome virus. BMC Biotechnol 2018;18.
    doi: 10.1186/s12896-018-0483-5pmc: PMC6240198pubmed: 30445953google scholar: lookup
  25. Hardmeier I, Schoster A. Association of EHV-1 antibody concentrations with shedding of EHV-1 in healthy horses and horses with EHV-1 infections. University of Zürich 2018. Aug.
  26. Crabb BS, Macpherson CM, Reubel GH, Browning GF, Studdert MJ, Drummer HE. A type-specific serological test to distinguish antibodies to equine herpesviruses 4 and 1. Arch Virol 1995.
    doi: 10.1007/BF01309860pubmed: 7710353google scholar: lookup
  27. Crabb BS, Studdert MJ. Epitopes of Glycoprotein G of Equine Herpesviruses 4 and 1 Located near the C Termini Elicit Type-Specific Antibody Responses in the Natural Host. J Virol 1993.
    pmc: PMC238063pubmed: 7690425
  28. Jumper J, Evans R, Pritzel A, Green T, Figurnov M, Ronneberger O. Highly accurate protein structure prediction with AlphaFold. Nature 2021;596: 583–589.
    doi: 10.1038/s41586-021-03819-2pmc: PMC8371605pubmed: 34265844google scholar: lookup
  29. Mirdita M, Schütze K, Moriwaki Y, Heo L, Ovchinnikov S, Steinegger M. ColabFold: making protein folding accessible to all. Nat Methods 2022;19: 679–682.
    doi: 10.1038/s41592-022-01488-1pmc: PMC9184281pubmed: 35637307google scholar: lookup
  30. Meng EC, Goddard TD, Pettersen EF, Couch GS, Pearson ZJ, Morris JH. UCSF ChimeraX: Tools for structure building and analysis. Protein Science 2023;32.
    doi: 10.1002/pro.4792pmc: PMC10588335pubmed: 37774136google scholar: lookup
  31. Vetter J, Papa G, Seyffert M, Gunasekera K, De Lorenzo G, Wiesendanger M. Rotavirus Spike Protein VP4 Mediates Viroplasm Assembly by Association to Actin Filaments. J Virol 2022;96.
    doi: 10.1128/jvi.01074-22pmc: PMC9472636pubmed: 35938869google scholar: lookup
  32. Damerval C, De Vienne D, Zivy M, Thiellement H. Technical improvements in two-dimensional electrophoresis increase the level of genetic variation detected in wheat-seedling proteins. Electrophoresis 1986;7: 52–54.
    doi: 10.1002/elps.1150070108google scholar: lookup
  33. Fulton KM, Twine SM. Immunoproteomics. 2nd ed. Fulton K, Twine S, editors. Humana New York, NY; 2020.
  34. Kremling V. Structural insights into ligand-receptor interaction of Equine herpesvirus type 1 and 4. Inaugural-Dissertation 2020.
  35. El Brini Z, Fassi Fihri O, Paillot R, Lotfi C, Amraoui F, El Ouadi H. Seroprevalence of equine herpesvirus 1 (EHV‐1) and equine herpesvirus 4 (EHV‐4) in the northern Moroccan horse populations. Animals 2021;11.
    doi: 10.3390/ani11102851pmc: PMC8532652pubmed: 34679874google scholar: lookup
  36. Gilkerson JR, Whalley JM, Drummer HE, Studdert MJ, Love DN. Epidemiological studies of equine herpesvirus 1 (EHV-1) in Thoroughbred foals: a review of studies conducted in the Hunter Valley of New South Wales between 1995 and 1997. 1997.
    pubmed: 10501158
  37. Ataseven VS, Daǧalp SB, Güzel M, Başaran Z, Tan MT, Geraghty B. Prevalence of equine herpesvirus-1 and equine herpesvirus-4 infections in equidae species in Turkey as determined by ELISA and multiplex nested PCR. Res Vet Sci 2009;86: 339–344.
    doi: 10.1016/j.rvsc.2008.06.001pubmed: 18649902google scholar: lookup
  38. Dunowska M, Gopakumar G, Perrott MR, Kendall AT, Waropastrakul S, Hartley CA. Virological and serological investigation of Equid herpesvirus 1 infection in New Zealand. Vet Microbiol 2015;176: 219–228.
    doi: 10.1016/j.vetmic.2015.01.016pubmed: 25666453google scholar: lookup
  39. Wellington JE, Gooley AA, Love DN, Whalley JM. N-terminal sequence analysis of equine herpesvirus 1 glycoproteins D and B and evidence for internal cleavage of the gene 71 product. Journal of General Virology 1996;77: 75–82.
    doi: 10.1099/0022-1317-77-1-75pubmed: 8558130google scholar: lookup
  40. Wagner B, Goodman LB, Babasyan S, Freer H, Torsteinsdóttir S, Svansson V. Antibody and cellular immune responses of naïve mares to repeated vaccination with an inactivated equine herpesvirus vaccine. Vaccine 2015;33: 5588–5597.
    doi: 10.1016/j.vaccine.2015.09.009pubmed: 26384446google scholar: lookup
  41. Allkofer A GM, RE LR, RM LG, WC VM, CA. Primary vaccination in foals: a comparison of the serological response to equine influenza and equine herpesvirus vaccines administered concurrently or 2 weeks apart. Arch Virol 2021;166: 571–579.
    pubmed: 33410993
  42. Vatti A MDM, PY CC, AJM GME. Original antigenic sin: A comprehensive review. J Autoimmun 2017;83: 12–21.
    doi: 10.1016/j.jaut.2017.04.008pubmed: 28479213google scholar: lookup

Citations

This article has been cited 1 times.
  1. Kambayashi Y, Bannai H, Nemoto M, Kawanishi N, Niwa H, Tsujimura K. Comparative analysis of 3 qPCR primer-probe sets for the detection of equid alphaherpesvirus 1. J Vet Diagn Invest 2026 Jan;38(1):77-83.
    doi: 10.1177/10406387251379857pubmed: 41055561google scholar: lookup
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