IgE ELISA using antisera derived from epsilon chain antigenic peptides detects allergen-specific IgE in allergic horses.
Abstract: Equine disease with an allergic etiology is common. Environmental antigens most often implicated as allergens in horses include molds, dusty hay, grass pollen, hay dust mites, and insect saliva. Although intradermal testing with allergen is a useful diagnostic tool for some species, skin testing frequently produces false positive results in horses. Allergen deprivation as a diagnostic tool is often impossible and at best it is ineffective at diagnosing the specific allergic reactivity. Synthesis of IgE after exposure to allergen is the instigator of the allergic process. While IgE exerts its effect after binding strongly to mast cell Fc receptors, the presence of free IgE in the serum can be used to quantify and determine the allergen specificity of the allergic disease. A lack of widely available reagents for detection of equine IgE has limited this approach in horses. We have used the nucleotide sequence of equine IgE to prepare a peptide-based immunogen to elicit equine epsilon chain-specific antisera. Selection of peptides was based on antigenic attributes of the deduced amino acid sequence of the equine epsilon chain. Six peptides were selected for conjugation to carrier molecules and rabbit immunization. Of these, one peptide elicited antisera that was successfully used in enzyme linked immunosorbant assay (ELISA) to screen horse serum from 64 allergic horses for allergen-specific IgE. Twenty-four of the 64 horses showed positive reactivity to one or more of the following allergens: grass, grain mill dust, mosquito, and horsefly. This study demonstrates the usefulness of peptide-based immunogens for development of antisera to rare or difficult to purify antigens such as IgE. Resultant antisera has great usefulness in diagnostic assays for equine allergy and as a research tool.
Publication Date: 2003-05-06 PubMed ID: 12730014DOI: 10.1016/s0165-2427(03)00034-5Google Scholar: Lookup
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- Journal Article
Summary
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This study demonstrates the development and successful use of peptide-based immunogens in creating antisera that can detect allergen-specific IgE in the serum of allergic horses, thus enhancing the diagnostic capabilities for equine allergies.
About the Study
- The research investigates an alternative diagnostic method to better detect and quantify allergic reactions in horses. Traditional skin testing often results in inaccurate readings.
- The researchers created a “peptide-based immunogen” (essentially, a specific triggering agent) using the DNA sequence of equine IgE to generate equine-specific antisera.
- This antisera was then used effectively in an enzyme-linked-immunosorbant assay (ELISA) to test horse serum for the presence of specific allergen-reactive IgEs.
Diagnostic Processes and Tools Used
- The researchers based the selection of peptides for this project on the antigenic characteristics of the amino acid sequence in the equine epsilon chain.
- Out of six peptides selected for conjugation to carrier molecules and rabbit immunization, one peptide triggered the production of useful antisera.
- Equine blood samples from 64 allergic horses were then screened using this antisera in ELISA testing. This particular type of test can identify and quantify substances that produce an immune response in the body.
Outcome of the Study
- The ELISA testing positively identified specific IgE reactions in twenty-four of the sixty-four horse samples tested. Reactive allergens included grass, grain mill dust, mosquito, and horsefly.
- This indicates the presence of these specific allergens in the horses’ environment that are causing an allergic response.
- The study provided proof of concept that this approach can successfully diagnose specific allergens causing health problems for horses. Hence, it could be a better diagnostic tool than the currently widely-used methods such as intradermal testing and allergen deprivation.
Implications of the Study
- This study establishes the usefulness of peptide-based immunogens in creating antisera for diagnostic purposes within veterinary medicine, particularly for detecting allergy-causing antigens.
- The success of this approach has the potential to improve allergen testing in equine veterinary care. It would make it easier to diagnose and then treat specific allergies in the horse population.
- Further, the antisera developed could be a significant tool for allergy research in horses.
Cite This Article
APA
Kalina WV, Pettigrew HD, Gershwin LJ.
(2003).
IgE ELISA using antisera derived from epsilon chain antigenic peptides detects allergen-specific IgE in allergic horses.
Vet Immunol Immunopathol, 92(3-4), 137-147.
https://doi.org/10.1016/s0165-2427(03)00034-5 Publication
Researcher Affiliations
- Department of Pathology, Microbiology, and Immunology, School of Veterinary Medicine, University of California, One Shields Ave., Davis, CA 95616, USA.
MeSH Terms
- Allergens / immunology
- Amino Acid Sequence
- Animals
- Antibodies, Monoclonal
- Antibody Specificity
- Blotting, Western / veterinary
- Enzyme-Linked Immunosorbent Assay / methods
- Enzyme-Linked Immunosorbent Assay / veterinary
- Female
- Horse Diseases / diagnosis
- Horse Diseases / immunology
- Horses
- Immunoglobulin E / blood
- Immunoglobulin E / immunology
- Male
- Molecular Sequence Data
- Peptide Fragments / immunology
- Rabbits
- Respiratory Hypersensitivity / diagnosis
- Respiratory Hypersensitivity / immunology
- Respiratory Hypersensitivity / veterinary
Citations
This article has been cited 2 times.- Mueller RS, Janda J, Jensen-Jarolim E, Rhyner C, Marti E. Allergens in veterinary medicine. Allergy 2016 Jan;71(1):27-35.
- Jensen-Jarolim E, Einhorn L, Herrmann I, Thalhammer JG, Panakova L. Pollen Allergies in Humans and their Dogs, Cats and Horses: Differences and Similarities. Clin Transl Allergy 2015;5:15.
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