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A histamine release assay to identify sensitization to Culicoides allergens in horses with skin hypersensitivity.
Abstract: Skin hypersensitivity is an allergic disease induced in horses by allergens of Culicoides midges. The condition is typically diagnosed by clinical signs and in some horses in combination with allergy testing such as intradermal skin testing or serological allergen-specific IgE determination. Here, we describe an alternative method for allergy testing: a histamine release assay (HRA) that combines the functional aspects of skin testing with the convenience of submitting a blood sample. The assay is based on the principle that crosslinking of allergen-specific IgE bound via high-affinity IgE receptors to the surfaces of mast cells and basophils induces the release of inflammatory mediators. One of these mediators is histamine. The histamine was then detected by a colorimetric enzyme-linked immunosorbent assay. The histamine assay was used to test 33 horses with skin hypersensitivity and 20 clinically healthy control animals for histamine release from their peripheral blood basophils after stimulation with Culicoides allergen extract or monoclonal anti-IgE antibody. An increased histamine release was observed in the horses with skin hypersensitivity compared to the control group after allergen-specific stimulation with Culicoides extract (p=0.023). In contrast, stimulation with anti-IgE induced similar amounts of released histamine in both groups (p=0.46). For further evaluation of the HRA, we prepared a receiver operating-characteristic (ROC) curve and performed a likelihood-ratio analysis for assay interpretation. Our results suggested that the assay is a valuable diagnostic tool to identify sensitization to Culicoides allergens in horses. Because some of the clinically healthy horses also showed sensitization to Culicoides extract, the assay cannot be used to distinguish allergic from non-allergic animals. The observation that sensitization is sometimes detectable in non-affected animals suggested that clinically healthy horses use immune mechanisms to control the reaction to Culicoides allergens that are different or absent in allergic horses.
Publication Date: 2008-09-11 PubMed ID: 18926574DOI: 10.1016/j.vetimm.2008.09.001Google 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.
- Comparative Study
- Journal Article
Summary
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This research describes a new method for diagnosing allergy in horses using a histamine release assay. This assay combines the functionality of skin testing with ease akin to taking a blood sample. Although this test cannot differentiate between allergic and non-allergic animals, it is effective in identifying sensitivity to specific allergens in horses.
What is the research about?
This study focuses on developing a new diagnostic tool, the histamine release assay (HRA), that combines convenience and functionality for identifying sensitization to Culicoides allergens in horses.
- The main concern of the research is a medical condition found in horses, known as skin hypersensitivity, an allergic disease triggered by allergens of Culicoides midges.
- The current diagnostic methods for this disease are by observing clinical signs and some horses are also subjected to allergy testing such as intradermal skin testing or serological allergen-specific IgE determination.
- This research aims to provide a more convenient and effective method to detect allergen-specific sensitivity in horses.
The Histamine Release Assay (HRA)
This method builds upon the principle that when allergen-specific IgE binds to mast cells and basophils via high-affinity IgE receptors, this induces the release of inflammatory mediators such as histamine.
- The histamine release can then be measured using a colorimetric enzyme-linked immunosorbent assay.
- The test subjects included 33 horses with skin hypersensitivity and 20 healthy horses. The peripheral blood basophils of these horses were stimulated with Culicoides allergen extract or monoclonal anti-IgE antibody.
- Increased histamine release was observed in hypersensitive horses compared to the control group after allergen-specific stimulation with Culicoides extract.
Evaluating the Efficacy of HRA
The effectiveness of HRA as a diagnostic tool was evaluated by preparing a receiver operating-characteristic (ROC) curve and performing a likelihood-ratio analysis.
- The results indicated that the assay is an effective diagnostic tool for identifying sensitization to Culicoides allergens in horses.
- However, the assay was unable to differentiate between allergic and non-allergic animals as some clinically healthy horses also showed sensitization towards Culicoides extract.
- This enforces the observation that healthy horses must use different or absent immune mechanisms to control the reaction to Culicoides allergens compared to allergic horses.
Cite This Article
APA
Wagner B, Childs BA, Erb HN.
(2008).
A histamine release assay to identify sensitization to Culicoides allergens in horses with skin hypersensitivity.
Vet Immunol Immunopathol, 126(3-4), 302-308.
https://doi.org/10.1016/j.vetimm.2008.09.001 Publication
Researcher Affiliations
- Department of Population Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853, USA. bw73@cornell.edu
MeSH Terms
- Animals
- Ceratopogonidae / immunology
- Histamine / immunology
- Horse Diseases / diagnosis
- Horse Diseases / immunology
- Horses
- Hypersensitivity / diagnosis
- Hypersensitivity / immunology
- Hypersensitivity / veterinary
- Likelihood Functions
- ROC Curve
- Skin Diseases / diagnosis
- Skin Diseases / immunology
- Skin Diseases / veterinary
Citations
This article has been cited 12 times.- Simonin EM, Wagner B. IgE-binding monocytes upregulate the coagulation cascade in allergic horses. Genes Immun 2023 Jun;24(3):130-138.
- Simonin EM, Babasyan S, Tarsillo J, Wagner B. IgE+ plasmablasts predict the onset of clinical allergy. Front Immunol 2023;14:1104609.
- Stammwitz V, Honnens Ä, Hochhuth D, Schuberth HJ. Increase of Adverse Events After Intravenous Injection of Gentamicin in Horses Between 2015 and 2017-From Marketing Authorization Holder's Point of View. Front Vet Sci 2021;8:710571.
- Raza F, Babasyan S, Larson EM, Freer HS, Schnabel CL, Wagner B. Peripheral blood basophils are the main source for early interleukin-4 secretion upon in vitro stimulation with Culicoides allergen in allergic horses. PLoS One 2021;16(5):e0252243.
- Larson EM, Babasyan S, Wagner B. IgE-Binding Monocytes Have an Enhanced Ability to Produce IL-8 (CXCL8) in Animals with Naturally Occurring Allergy. J Immunol 2021 May 15;206(10):2312-2321.
- Novotny EN, White SJ, Wilson AD, Stefánsdóttir SB, Tijhaar E, Jonsdóttir S, Frey R, Reiche D, Rose H, Rhyner C, Schüpbach-Regula G, Torsteinsdóttir S, Alcocer M, Marti E. Component-resolved microarray analysis of IgE sensitization profiles to Culicoides recombinant allergens in horses with insect bite hypersensitivity. Allergy 2021 Apr;76(4):1147-1157.
- Raza F, Ivanek R, Freer H, Reiche D, Rose H, Torsteinsdóttir S, Svansson V, Björnsdóttir S, Wagner B. Cul o 2 specific IgG3/5 antibodies predicted Culicoides hypersensitivity in a group imported Icelandic horses. BMC Vet Res 2020 Aug 10;16(1):283.
- Larson EM, Babasyan S, Wagner B. Phenotype and function of IgE-binding monocytes in equine Culicoides hypersensitivity. PLoS One 2020;15(5):e0233537.
- François L, Hoskens H, Velie BD, Stinckens A, Tinel S, Lamberigts C, Peeters L, Savelkoul HFJ, Tijhaar E, Lindgren G, Janssens S, Ducro BJ, Buys N, Schurink AA. Genomic Regions Associated with IgE Levels against Culicoides spp. Antigens in Three Horse Breeds. Genes (Basel) 2019 Aug 8;10(8).
- Torsteinsdottir S, Scheidegger S, Baselgia S, Jonsdottir S, Svansson V, Björnsdottir S, Marti E. A prospective study on insect bite hypersensitivity in horses exported from Iceland into Switzerland. Acta Vet Scand 2018 Nov 3;60(1):69.
- Lavergne SN. In Vitro Research Tools in the Field of Human Immediate Drug Hypersensitivity and Their Present Use in Small Animal Veterinary Medicine. Vet Sci 2016 Dec 22;4(1).
- Simonin EM, Torsteinsdóttir S, Svansson V, Björnsdóttir S, Freer H, Tarsillo J, Wagner B. Early allergen introduction overrides allergy predisposition in offspring of horses with Culicoides hypersensitivity. Front Immunol 2025;16:1654693.
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