Monoclonal antibodies to equine CD23 identify the low-affinity receptor for IgE on subpopulations of IgM+ and IgG1+ B-cells in horses.
Abstract: CD23, also called FcεRII, is the low-affinity receptor for IgE and has first been described as a major receptor regulating IgE responses. In addition, CD23 also binds to CD21, integrins and MHC class II molecules and thus has a much wider functional role in immune regulation ranging from involvement in antigen-presentation to multiple cytokine-like functions of soluble CD23. The role of CD23 during immune responses of the horse is less well understood. Here, we expressed equine CD23 in mammalian cells using a novel IL-4 expression system. Expression resulted in high yield of recombinant IL-4/CD23 fusion protein which was purified and used for the generation of monoclonal antibodies (mAbs) to equine CD23. Seven anti-CD23 mAbs were further characterized. The expression of the low-affinity IgE receptor on equine peripheral blood mononuclear cells was analyzed by flow cytometric analysis. Cell surface staining showed that CD23 is mainly expressed by a subpopulation of equine B-cells. Only a very few equine T-cells or monocytes expressed CD23. CD23(+) B-cells were either IgM(+) or IgG1(+) cells. All CD23(+) cells were also positive for cell surface IgE staining suggesting in vivo IgE binding by the receptor. Two of the CD23 mAbs detected either the complete extracellular region of CD23 or a 22kDa cleavage product of CD23 by Western blotting. The new anti-CD23 mAbs provide valuable reagents to further analyze the roles of CD23 during immune responses of the horse in health and disease.
Copyright © 2012 Elsevier B.V. All rights reserved.
Publication Date: 2012-02-18 PubMed ID: 22405681DOI: 10.1016/j.vetimm.2012.02.007Google Scholar: Lookup
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- Journal Article
- Research Support
- U.S. Gov't
- Non-P.H.S.
Summary
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This research explores the less understood role of CD23, a receptor that regulates IgE responses, during immune responses in horses. The study generated monoclonal antibodies to equine CD23 using a novel IL-4 expression system and concluded that these new anti-CD23 antibodies can be useful for further research into the role of CD23 in equine health and disease.
Research Objective and Process
- The research aimed to understand the role of CD23 during immune responses in horses, a role that is not well understood.
- The researchers used a novel Interleukin-4 (IL-4) expression system to express equine CD23 in mammalian cells. Interleukin-4 is a crucial protein involved in immune responses.
- The expression resulted in a high yield of the receptor protein CD23, in fusion with IL-4, which was subsequently purified.
- This purified protein was used to generate monoclonal antibodies towards equine CD23. Monoclonal antibodies are identical immune cells that are clones of a unique parent cell, and they target one type of cell.
- These manufactured monoclonal antibodies were characterized, and seven anti-CD23 antibodies were further studied.
Results and Findings
- The research found that CD23 is mainly expressed by a subpopulation of equine B-cells. B-cells are a type of white blood cell that creates antibodies against antigens.
- A very few T-cells or monocytes, other types of immune cells, expressed CD23.
- CD23 positive B-cells were either Immunoglobulin M (IgM) or Immunoglobulin G (IgG1) cells. IgM and IgG1 are types of antibodies involved in response to infection.
- All CD23 positive cells also tested positive for cell surface Immunoglobulin E (IgE) staining, suggesting in vivo IgE binding by the receptor.
- Two of the CD23 monoclonal antibodies detected either the complete extracellular region of CD23 or a 22 kiloDalton (kDa) cleavage product of CD23 by Western blotting, a common analytical technique used to detect specific proteins in a sample.
Conclusions and Implications
- The findings provide valuable tools to further analyze CD23’s role during immune responses in horses.
- Understanding the role of CD23 could have implications for improving horse health and managing disease.
Cite This Article
APA
Wagner B, Hillegas JM, Babasyan S.
(2012).
Monoclonal antibodies to equine CD23 identify the low-affinity receptor for IgE on subpopulations of IgM+ and IgG1+ B-cells in horses.
Vet Immunol Immunopathol, 146(2), 125-134.
https://doi.org/10.1016/j.vetimm.2012.02.007 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
- Antibodies, Monoclonal / genetics
- Antibodies, Monoclonal / immunology
- B-Lymphocytes / immunology
- Blotting, Western / veterinary
- Cloning, Molecular / methods
- Flow Cytometry / veterinary
- Horses / blood
- Horses / immunology
- Immunoglobulin E / immunology
- Immunoglobulin G / immunology
- Immunoglobulin M / immunology
- Leukocytes, Mononuclear / immunology
- Receptors, IgE / immunology
- Recombinant Fusion Proteins / genetics
- Recombinant Fusion Proteins / immunology
Citations
This article has been cited 17 times.- Simonin EM, Babasyan S, Tarsillo J, Wagner B. IgE+ plasmablasts predict the onset of clinical allergy.. Front Immunol 2023;14:1104609.
- Sipka A, Babasyan S, Mann S, Freer H, Klaessig S, Wagner B. Development of monoclonal antibodies for quantification of bovine tumor necrosis factor-α.. JDS Commun 2021 Nov;2(6):415-420.
- Mair KH, Crossman AJ, Wagner B, Babasyan S, Noronha L, Boyd P, Zarlenga D, Stadler M, van Dongen KA, Gerner W, Saalmüller A, Lunney JK. The Natural Cytotoxicity Receptor NKp44 (NCR2, CD336) Is Expressed on the Majority of Porcine NK Cells Ex Vivo Without Stimulation.. Front Immunol 2022;13:767530.
- Guarino C, Larson E, Babasyan S, Rollins A, Joshi LR, Laverack M, Parrilla L, Plocharczyk E, Diel DG, Wagner B. Development of a quantitative COVID-19 multiplex assay and its use for serological surveillance in a low SARS-CoV-2 incidence community.. PLoS One 2022;17(1):e0262868.
- Kang H, Bienzle D, Lee GKC, Piché É, Viel L, Odemuyiwa SO, Beeler-Marfisi J. Flow cytometric analysis of equine bronchoalveolar lavage fluid cells in horses with and without severe equine asthma.. Vet Pathol 2022 Jan;59(1):91-99.
- Fasanello DC, Su J, Deng S, Yin R, Colville MJ, Berenson JM, Kelly CM, Freer H, Rollins A, Wagner B, Rivas F, Hall AR, Rahbar E, DeAngelis PL, Paszek MJ, Reesink HL. Hyaluronic acid synthesis, degradation, and crosslinking in equine osteoarthritis: TNF-α-TSG-6-mediated HC-HA formation.. Arthritis Res Ther 2021 Aug 20;23(1):218.
- Watkins A, Fasanello D, Stefanovski D, Schurer S, Caracappa K, D'Agostino A, Costello E, Freer H, Rollins A, Read C, Su J, Colville M, Paszek M, Wagner B, Reesink H. Investigation of synovial fluid lubricants and inflammatory cytokines in the horse: a comparison of recombinant equine interleukin 1 beta-induced synovitis and joint lavage models.. BMC Vet Res 2021 May 12;17(1):189.
- 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.
- Palmer MV, Martins M, Falkenberg S, Buckley A, Caserta LC, Mitchell PK, Cassmann ED, Rollins A, Zylich NC, Renshaw RW, Guarino C, Wagner B, Lager K, Diel DG. Susceptibility of white-tailed deer (Odocoileus virginianus) to SARS-CoV-2.. J Virol 2021 May 10;95(11).
- Patel RS, Tomlinson JE, Divers TJ, Van de Walle GR, Rosenberg BR. Single-cell resolution landscape of equine peripheral blood mononuclear cells reveals diverse cell types including T-bet(+) B cells.. BMC Biol 2021 Jan 22;19(1):13.
- Larson EM, Babasyan S, Wagner B. Phenotype and function of IgE-binding monocytes in equine Culicoides hypersensitivity.. PLoS One 2020;15(5):e0233537.
- Armstrong C, Cassimeris L, Da Silva Santos C, Micoogullari Y, Wagner B, Babasyan S, Brooks S, Galantino-Homer H. The expression of equine keratins K42 and K124 is restricted to the hoof epidermal lamellae of Equus caballus.. PLoS One 2019;14(9):e0219234.
- Wimer CL, Schnabel CL, Perkins G, Babasyan S, Freer H, Stout AE, Rollins A, Osterrieder N, Goodman LB, Glaser A, Wagner B. The deletion of the ORF1 and ORF71 genes reduces virulence of the neuropathogenic EHV-1 strain Ab4 without compromising host immunity in horses.. PLoS One 2018;13(11):e0206679.
- Schnabel CL, Wimer CL, Perkins G, Babasyan S, Freer H, Watts C, Rollins A, Osterrieder N, Wagner B. Deletion of the ORF2 gene of the neuropathogenic equine herpesvirus type 1 strain Ab4 reduces virulence while maintaining strong immunogenicity.. BMC Vet Res 2018 Aug 22;14(1):245.
- Wagner B, Perkins G, Babasyan S, Freer H, Keggan A, Goodman LB, Glaser A, Torsteinsdóttir S, Svansson V, Björnsdóttir S. Neonatal Immunization with a Single IL-4/Antigen Dose Induces Increased Antibody Responses after Challenge Infection with Equine Herpesvirus Type 1 (EHV-1) at Weanling Age.. PLoS One 2017;12(1):e0169072.
- Noronha LE, Harman RM, Wagner B, Antczak DF. Generation and characterization of monoclonal antibodies to equine NKp46.. Vet Immunol Immunopathol 2012 Jun 15;147(1-2):60-8.
- Noronha LE, Harman RM, Wagner B, Antczak DF. Generation and characterization of monoclonal antibodies to equine CD16.. Vet Immunol Immunopathol 2012 Apr 15;146(2):135-42.
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