Development of a Comprehensive Protein Microarray for Immunoglobulin E Profiling in Horses with Severe Equine Asthma (sEA)

White, S, Moore-Colyer, M, Marti, E, Couëtil, L, Hannant, D, Richard, E and Alcocer, M (2019) Development of a Comprehensive Protein Microarray for Immunoglobulin E Profiling in Horses with Severe Equine Asthma (sEA). Journal of Veterinary and Internal Medicine. ISSN 1939-1676

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Abstract

Abstract: Background - Severe equine asthma (sEA) is a prevalent, performance-limiting disease associated with increased allergen-specific immunoglobulin E (IgE) against a range of environmental aeroallergen. Objective – To develop a protein microarray platform to profile IgE against a range of proven and novel environmental proteins in sEA-affected horses. Animals - Six sEA-affected and six clinically healthy warmblood performance horses. Methods - A protein microarray (n=384) was developed using protein extracts and purified proteins from a large number of families including pollen, bacteria, fungi and arthropods associated with the equine 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 revealing an association between sEA and pollen sensitization. Immunolabeling methods confirmed the accuracy of a commercially available mouse anti-horse IgE 3H10 source (R2=.91). Biological fluid source evaluation demonstrated that sera and bronchoalveolar lavage fluid (BALF) yielded the same specific IgE profile (average R2=.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 (R2=.97), as was specificity via protein inhibition assays. As expected, arthropods, fungi and pollens have shown the greatest discrimination for sEA. Conclusion - Here we have established that protein microarrays can be used for large scale IgE mapping of allergens associated with the equine environment. This technology provides a sound platform for specific diagnosis, management and treatment of sEA.

Item Type: Article
Keywords: severe equine asthma, horse, protein microarray, IgE, allergen
Divisions: Equine Management and Science
Depositing User: Prof Meriel Moore-Colyer
Date Deposited: 10 Jun 2019 15:04
Last Modified: 13 Sep 2019 13:38
URI: http://rau.repository.guildhe.ac.uk/id/eprint/16146

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