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Home / Equine Research Bank / J Vis Exp / Development of an in vitro model system for studying the int...
Journal of visualized experiments : JoVE2014; (93); e52222; doi: 10.3791/52222

Development of an in vitro model system for studying the interaction of Equus caballus IgE with its high-affinity receptor FcεRI.

Abstract: The interaction of IgE with its high-affinity Fc receptor (FcεRI) followed by an antigenic challenge is the principal pathway in IgE mediated allergic reactions. As a consequence of the high affinity binding between IgE and FcεRI, along with the continuous production of IgE by B cells, allergies usually persist throughout life, with currently no permanent cure available. Horses, especially race horses, which are commonly inbred, are a species of mammals that are very prone to the development of hypersensitivity responses, which can seriously affect their performance. Physiological responses to allergic sensitization in horses mirror that observed in humans and dogs. In this paper we describe the development of an in situ assay system for the quantitative assessment of the release of mediators of the allergic response pertaining to the equine system. To this end, the gene encoding equine FcεRIα was transfected into and expressed onto the surface of parental Rat Basophil Leukemia (RBL-2H3.1) cells. The gene product of the transfected equine α-chain formed a functional receptor complex with the endogenous rat β- and γ-chains. The resultant assay system facilitated an assessment of the quantity of mediator secreted from equine FcεRIα transfected RBL-2H3.1 cells following sensitization with equine IgE and antigenic challenge using β-hexosaminidase release as a readout. Mediator release peaked at 36.68% ± 4.88% at 100 ng ml(-1) of antigen. This assay was modified from previous assays used to study human and canine allergic responses. We have also shown that this type of assay system has multiple applications for the development of diagnostic tools and the safety assessment of potential therapeutic intervention strategies in allergic disease.
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Publication Date: 2014-11-01 PubMed ID: 25406512PubMed Central: PMC4541457DOI: 10.3791/52222Google 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.

This research focuses on understanding the interaction of IgE antibodies with their receptor, FcεRI, in horses, an issue central to allergic reactions. The scientists developed an in vitro assay system to assess the release of mediators in allergic reactions and propose potential uses for diagnosis and treatment of allergies in horses.

Understanding Equine Allergies

  • This study specifically focuses on IgE mediated allergic reactions in horses. Just like humans and canines, horses often suffer from allergies that are caused due to the interaction of IgE antibodies with their high-affinity Fc receptor, FcεRI with antigenic challenge.
  • Persistent allergies in horses can crucially affect their performance. Therefore, the development of hypersensitivity responses, specifically in race horses, is a matter of concern.

Development of an In Vitro Assay System

  • The researchers transformed Rat Basophil Leukemia (RBL-2H3.1) cells, a commonly used cellular model for studying allergic reactions, to express the horse FcεRIα receptor. This receptor was then used to form a functional complex with the rat β- and γ-chains.
  • The assay system facilitated an assessment of the mediator secreted from the transfected cells after sensitization with horse IgE and antigenic challenge. β-hexosaminidase release was used as a readout to quantify this mediator release.
  • The mediator release peaked at 36.68% ± 4.88% at 100 ng ml(-1) of antigen, indicating a measurable response.

Advancements from Previously Used Assays

  • The assay system developed in this study was a significant modification and advancement over previous assays used to study human and canine allergic responses.
  • It specifically allowed for the study of equine allergic reactions, providing a vital step in understanding the exact dynamics involved in equine allergies.

Implications and Applications

  • The development of this assay system comes with multiple applications. It could be potentially applied for the development of diagnostic tools that are adapted for assessing allergies in horses.
  • Test results from this assay could provide useful insights for the safety assessment of potential therapeutic intervention strategies in allergic disease. This could significantly improve our ability to treat and even prevent allergies in horses, thereby enhancing their performance.

Cite This Article

APA
Sabban S, Ye H, Helm B. (2014). Development of an in vitro model system for studying the interaction of Equus caballus IgE with its high-affinity receptor FcεRI. J Vis Exp(93), e52222. https://doi.org/10.3791/52222

Publication

Journal of visualized experiments : JoVE
ISSN: 1940-087X
NlmUniqueID: 101313252
Country: United States
Language: English
Issue: 93
Pages: e52222
PII: 52222

Researcher Affiliations

Sabban, Sari
  • Biological Department, King Abdulaziz University; sari.sabban@gmail.com.
Ye, Hongtu
  • The Krebs Institute for Biomolecular Research, Department of Molecular Biology and Biotechnology, The University of Sheffield.
Helm, Birgit
  • The Krebs Institute for Biomolecular Research, Department of Molecular Biology and Biotechnology, The University of Sheffield.

MeSH Terms

  • Animals
  • Antigens / immunology
  • Cell Line, Tumor
  • Dogs
  • Horses
  • Humans
  • Hypersensitivity / genetics
  • Hypersensitivity / immunology
  • Immunoglobulin E / immunology
  • Leukemia, Basophilic, Acute / genetics
  • Leukemia, Basophilic, Acute / immunology
  • Rats
  • Receptors, IgE / genetics
  • Receptors, IgE / immunology
  • Transfection
  • beta-N-Acetylhexosaminidases / immunology
  • beta-N-Acetylhexosaminidases / metabolism

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Citations

This article has been cited 3 times.
  1. Sabban SS. Computationally grafting an IgE epitope onto a scaffold: Implications for a pan anti-allergy vaccine design.. Comput Struct Biotechnol J 2021;19:4738-4750.
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  2. Spindel S, Sapsford KE. Evaluation of optical detection platforms for multiplexed detection of proteins and the need for point-of-care biosensors for clinical use.. Sensors (Basel) 2014 Nov 25;14(12):22313-41.
    doi: 10.3390/s141222313pubmed: 25429414google scholar: lookup
  3. Molinari G, Colombo G, Celenza C. Respiratory allergies: a general overview of remedies, delivery systems, and the need to progress.. ISRN Allergy 2014;2014:326980.
    doi: 10.1155/2014/326980pubmed: 25006500google scholar: lookup
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