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Home / Equine Research Bank / J Vet Intern Med / Development of a comprehensive protein microarray for immuno...
Journal of veterinary internal medicine2019; 33(5); 2327-2335; doi: 10.1111/jvim.15564

Development of a comprehensive protein microarray for immunoglobulin E profiling in horses with severe asthma.

Abstract: Severe asthma in horses, known as severe equine asthma (SEA), is a prevalent, performance-limiting disease associated with increased allergen-specific immunoglobulin E (IgE) against a range of environmental aeroallergens. Objective: To develop a protein microarray platform to profile IgE against a range of proven and novel environmental proteins in SEA-affected horses. Methods: Six SEA-affected and 6 clinically healthy Warmblood performance horses. Methods: Developed a protein microarray (n = 384) using protein extracts and purified proteins from a large number of families including pollen, bacteria, fungi, and arthropods associated with the horses, environment. Conditions were optimized and assessed for printing, incubation, immunolabeling, biological fluid source, concentration techniques, reproducibility, and specificity. Results: This method identified a number of novel allergens, while also identifying an association between SEA and pollen sensitization. Immunolabeling methods confirmed the accuracy of a commercially available mouse anti-horse IgE 3H10 source (R = 0.91). Biological fluid source evaluation indicated that sera and bronchoalveolar lavage fluid (BALF) yielded the same specific IgE profile (average R = 0.75). Amicon centrifugal filters were found to be the most efficient technique for concentrating BALF for IgE analysis at 40-fold. Overnight incubation maintained the same sensitization profile while increasing sensitivity. Reproducibility was demonstrated (R = 0.97), as was specificity using protein inhibition assays. Arthropods, fungi, and pollens showed the greatest discrimination for SEA. Conclusions: We have established that protein microarrays can be used for large-scale IgE mapping of allergens associated with the environment of horses. This technology provides a sound platform for specific diagnosis, management, and treatment of SEA.
© 2019 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of the American College of Veterinary Internal Medicine.
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Publication Date: 2019-08-20 PubMed ID: 31429513PubMed Central: PMC6766494DOI: 10.1111/jvim.15564Google 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 focuses on creating a comprehensive protein microarray platform for analyzing immunoglobulin E (IgE) allergies in horses exhibiting severe asthma or SEA. This method revealed new allergens and established an association between SEA and pollen sensitization in horses.

Research Objective and Methods

The primary goal of the study was to create a protein microarray platform that could profile IgE, which reacts to different environmental proteins in horses suffering from SEA. The researchers utilized protein extracts and purified proteins from diverse allergen families like pollen, bacteria, fungi, and arthropods found in the horses’ environment.

  • Six horses with SEA and six healthy Warmblood performance horses were used for the study.
  • The research team optimized and evaluated several parameters like printing, incubation, immunolabeling, biological fluid source, techniques for concentration, reproducibility, and specificity in the course of the research.

Results of the Research

The protein microarray method developed identified new allergens and linked SEA to pollen sensitization in horses.

  • The team confirmed the accuracy of a readily available mouse anti-horse IgE 3H10 source with immunolabeling methods.
  • Both serum and bronchoalveolar lavage fluid (BALF) gave the same specific IgE profile.
  • Amicon centrifugal filters were determined as the most effective method to concentrate BALF for IgE analysis.
  • An overnight incubation increased the sensitivity while keeping the sensitization profile the same.
  • Protein inhibition assays were used to demonstrate the method’s specificity, and reproducibility was also proven.
  • Arthropods, fungi, and pollens showed the most significant differentiation for SEA.

Conclusion

The study established that protein microarrays could effectively map large-scale IgE allergens associated with horses’ environment. This novel technology can provide a sturdy basis for the specific diagnosis, management, and treatment of SEA in horses.

Cite This Article

APA
White S, Moore-Colyer M, Marti E, Coüetil L, Hannant D, Richard EA, Alcocer M. (2019). Development of a comprehensive protein microarray for immunoglobulin E profiling in horses with severe asthma. J Vet Intern Med, 33(5), 2327-2335. https://doi.org/10.1111/jvim.15564

Publication

Journal of veterinary internal medicine
ISSN: 1939-1676
NlmUniqueID: 8708660
Country: United States
Language: English
Volume: 33
Issue: 5
Pages: 2327-2335

Researcher Affiliations

White, Samuel
  • School of Equine Management and Science, Royal Agricultural University, Gloucestershire, United Kingdom.
  • School of Biosciences, University of Nottingham, Loughborough, United Kingdom.
  • Animal and Equine Science, Nottingham Trent University, Nottinghamshire, United Kingdom.
Moore-Colyer, Meriel
  • School of Equine Management and Science, Royal Agricultural University, Gloucestershire, United Kingdom.
Marti, Eliane
  • Department of Clinical Research and Veterinary Public Health, University of Bern, Bern, Switzerland.
Coüetil, Laurent
  • Veterinary Clinical Sciences, College of Veterinary Medicine, Purdue University, West Lafayette, Indiana, USA.
Hannant, Duncan
  • School of Veterinary Medicine and Science, University of Nottingham, Loughborough, United Kingdom.
Richard, Eric A
  • LABÉO Frank Duncombe, Caen Cedex, France.
  • Normandie University, UniCaen, BIOTARGEN, Saint-Contest, France.
Alcocer, Marcos
  • School of Biosciences, University of Nottingham, Loughborough, United Kingdom.

MeSH Terms

  • Animals
  • Arthropods / immunology
  • Asthma / blood
  • Asthma / immunology
  • Asthma / veterinary
  • Bronchoalveolar Lavage Fluid / immunology
  • Case-Control Studies
  • Fungi / immunology
  • Horse Diseases / blood
  • Horse Diseases / immunology
  • Horses
  • Immunoglobulin E / blood
  • Immunoglobulin E / immunology
  • Mice
  • Pollen / immunology
  • Protein Array Analysis / methods
  • Protein Array Analysis / veterinary

Grant Funding

  • Fred And Marjorie Sainsbury Charitable Trust
  • HAYGAIN
  • The Royal Agricultural University

Conflict of Interest Statement

Authors declare no conflict of interest.

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Citations

This article has been cited 8 times.
  1. Wyler M, Sage SE, Marti E, White S, Gerber V. Protein microarray allergen profiling in bronchoalveolar lavage fluid and serum of horses with asthma. J Vet Intern Med 2023 Jan;37(1):328-337.
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  4. White SJ, Moore-Colyer M, Marti E, Hannant D, Gerber V, Coüetil L, Richard EA, Alcocer M. Antigen array for serological diagnosis and novel allergen identification in severe equine asthma. Sci Rep 2019 Oct 23;9(1):15170.
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  5. Dély S, Gerber V, Peters LM, Sage SE. Association between equine asthma and fungal elements in the tracheal wash: An environment-matched case-control study. PLoS One 2024;19(9):e0309835.
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  6. Wjst VF, Lübke S, Wagner B, Rhyner C, Jentsch MC, Arnold C, Lohmann KL, Schnabel CL. Aspergillus fumigatus antigen-reactive Th17 cells are enriched in bronchoalveolar lavage fluid in severe equine asthma. Front Immunol 2024;15:1367971.
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  7. Schnabel CL, Jentsch MC, Lübke S, Kaiser-Thom S, Gerber V, Vrtala S, Huang HJ, Rhyner C, Wagner B, Hoffmann R, Volke D. Immunoproteomics reveal increased serum IgG3/5 binding to Dermatophagoides and yeast protein antigens in severe equine asthma in a preliminary study. Front Immunol 2023;14:1293684.
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