Monoclonal anti-equine IgE antibodies with specificity for different epitopes on the immunoglobulin heavy chain of native IgE.
Abstract: In this study we describe the generation of monoclonal antibodies (mAbs), which recognize different epitopes of the equine IgE constant heavy chain. Equi-murine recombinant IgE (rIgE), composed of the murine V(H)186.2 heavy chain variable region, linked to the equine IgE constant heavy chain and expressed together with the murine lambda(1) chain in J558L cells was used to immunize BALB/C mice. A total of 17 different mAbs were obtained, which recognized the rIgE heavy chain constant region. None of the mAbs reacted with monoclonal equine isotypes IgM, IgG1 (IgGa), IgG3 (IgG(T)), IgG4 (IgGb) or isolated equine light chains, IgGc and IgA from horse serum, or the native mAb B1-8delta, expressing the same heavy chain variable regions and light chains. One of the mAbs (alphaIgE-132) recognized the recombinant equine IgE, but did not recognize any protein in equine serum, i.e. native IgE. A total of 16 mAbs detected a serum protein of approximately 210,000Da on Western blots, corresponding to the expected MW of native IgE. In addition, one of the mAbs (alphaIgE-176) detected a protein of 76,000Da under reducing conditions, most likely the equine IgE heavy chain. According to binding inhibition studies, the equine IgE specific mAbs recognize at least two different epitopes of the equine IgE. In an ELISA using two anti-IgE mAbs which recognized different epitopes, no significant differences in the concentration of total serum IgE could be detected between adult Icelandic horses with IgE-mediated type I allergy (summer eczema) and healthy control animals. In Icelandic horse foals, no serum IgE could be measured 6 months post partum. All anti-IgE mAbs recognized a small population (1.3+/-0.5%) of leukocytes from adult Icelandic horses by surface immunofluorescence, but no cells could be detected in foal blood. The stained leukocytes from adult horses could be enriched by magnetic cell sorting and contained 32% basophils, 53% monocytes and/or large lymphocytes, 13% small lymphocytes and 2% eosinophils.
Publication Date: 2003-03-12 PubMed ID: 12628763DOI: 10.1016/s0165-2427(03)00007-2Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This research involves the production of specific monoclonal antibodies against different sites on the constant heavy chain part of equine (horse) IgE, a type of immunoglobulin involved in allergic reactions. The study found no significant difference in total serum IgE concentrations between adult Icelandic horses suffering from type I allergy and healthy controls, and detected the presence of IgE on certain white blood cells in adult, but not young, horses.
Background and Methods
- The researchers utilized recombinant IgE (genetically engineered antibodies) made up of the heavy chain variable region from a mouse and the constant heavy chain from a horse IgE. This was expressed along with a particular type of mouse light chain in a mammalian cell line.
- They then vaccinated mice with this material, causing the mice’s immune system to manufacture antibodies against the horse IgE components of the recombinant IgE molecules.
- A total of 17 distinct monoclonal antibodies (mAbs) were produced, each of which recognizes different parts (epitopes) of the equine IgE constant heavy chain.
Findings
- None of these mAbs reacted with other types of horse antibodies (IgM, IgG1, IgG3, IgG4, IgGc, IgA) or the isolated light chains of these other antibodies.
- The researchers were able to use these mAbs to detect both native (natural, non-recombinant) horse IgE and the recombinant horse IgE they created.
- Binding inhibition studies showed that the mAbs recognize at least two different epitopes on the equine IgE heavy chain.
- An enzyme-linked immunoassay (ELISA) test found no significant differences in total IgE levels in the blood between adult horses with summer eczema (a type I allergy) and healthy horses.
- However, they found a small population of leukocytes (white blood cells) in adult horses that are recognized by these mAbs. These cells are believed to contain IgE on their surface.
Significance
- The monocyte and lymphocyte populations that are bound by anti-IgE mAbs could provide an important clue to understanding the role of this immunoglobulin in allergic reactions.
- Understanding the mechanism of IgE responses in horses may help veterinarians to better diagnose and treat equine allergies.
- These antibodies can also be used in the future to further study the structure and function of IgE in horses.
Cite This Article
APA
Wagner B, Radbruch A, Rohwer J, Leibold W.
(2003).
Monoclonal anti-equine IgE antibodies with specificity for different epitopes on the immunoglobulin heavy chain of native IgE.
Vet Immunol Immunopathol, 92(1-2), 45-60.
https://doi.org/10.1016/s0165-2427(03)00007-2 Publication
Researcher Affiliations
- Immunology Unit, Hannover School of Veterinary Medicine, Bischofsholer Damm 15, Germany. bw73@cornell.edu
MeSH Terms
- Animals
- Antibodies, Anti-Idiotypic / immunology
- Antibodies, Monoclonal / immunology
- Antibody Specificity / immunology
- Blotting, Western / veterinary
- Cross Reactions / immunology
- Enzyme-Linked Immunosorbent Assay / veterinary
- Epitopes / immunology
- Female
- Flow Cytometry / veterinary
- Horses / immunology
- Immunoglobulin E / immunology
- Immunoglobulin Heavy Chains / immunology
- Immunoglobulin Isotypes
- Immunomagnetic Separation / veterinary
- Leukocytes / immunology
- Mice
- Mice, Inbred BALB C
- Recombinant Proteins / immunology
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