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Home / Equine Research Bank / Vet Immunol Immunopathol / A monoclonal antibody to equine interleukin-4.
Veterinary immunology and immunopathology2006; 110(3-4); 363-367; doi: 10.1016/j.vetimm.2006.01.001

A monoclonal antibody to equine interleukin-4.

Abstract: Interleukin-4 (IL-4) is secreted by T helper type 2 cells, mast cells, basophils and eosinophils. Detection of IL-4 can contribute the evaluation of cellular immune responses during infectious diseases, immunological disorders or vaccination. We used recombinant equine IL-4 to generate a monoclonal antibody (mAb) to equine IL-4. The mAb detected recombinant IL-4 in mammalian cells transfected with different plasmids containing IL-4 cDNA. After mitogen stimulation of equine peripheral blood mononuclear cells, an intracellular protein was recognized by the new mAb in 1-2% of lymphocytes using flow cytometric analysis. In the presence of the secretion blocker Brefeldin A, the protein accumulated and was detected in 4-8% of lymphocytes stimulated with phorbol 12-myristate 13-acetate and ionomycin. Double staining with the new mAb and T-cell or B-cell markers identified a subpopulation of CD4+ T-cells expressing the protein recognized by the mAb. In addition, the protein was detectable in cell culture supernatants of mitogen stimulated cells by ELISA when using the new mAb for coating of the plates and a polyclonal antiserum to equine IL-4 for detection. In conclusion, the new mAb detects equine IL-4 and can be used for intracellular staining and ELISA to measure this important cytokine.
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Publication Date: 2006-02-09 PubMed ID: 16480777DOI: 10.1016/j.vetimm.2006.01.001Google Scholar: Lookup
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  • Journal Article
  • Research Support
  • Non-U.S. Gov't

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.

The research article describes the creation and testing of a monoclonal antibody for equine Interleukin-4 (IL-4), a protein secreted by certain immune cells. The team tested the antibody’s ability to detect equine IL-4 in different cells and situations, showing its potential use in studying immune responses.

Less Technical Explanation of the Topic

  • Interleukin-4 (IL-4) is a protein that plays a key role in the immune system, being produced by a variety of immune cells. Its level can provide insight into how the immune system is responding in different situations, such as during infections, immunological disorders, or following vaccinations.
  • The researchers in this study generated a monoclonal antibody — a type of lab-made protein designed to interact with a specific substance — for equine, or horse, IL-4.

Creating and Testing the Monoclonal Antibody

  • This new monoclonal antibody (mAb) was produced using a recombinant form of equine IL-4, a lab-modified version of the protein. This mAb was then used to detect IL-4 in different cells that had been manipulated to carry the gene for IL-4.
  • Focusing on one type of cell from the blood, called peripheral blood mononuclear cells, the researchers found that a new protein was marked by the mAb in a small fraction of these cells. This was done using a technique called flow cytometry, which can analyze individual particles, such as cells, in a fluid sample.

Understanding How the Monoclonal Antibody Reacts

  • By blocking protein secretion, they observed an increase in the accumulation of the protein recognized by the mAb. Moreover, using different cell markers, they found that a subset of T-cells, a type of white blood cell, expressed the protein recognized by the mAb.
  • This new antibody was also able to detect the IL-4 protein in the liquid surrounding cultured cells that had been stimulated with certain stimuli, as demonstrated by an enzyme-linked immunosorbent assay (ELISA) — a common lab tool for detecting the presence of a substance in a liquid.

Conclusions and Implications

  • The study concludes that this newly created mAb is capable of detecting equine IL-4 and could be potentially useful in both staining cells for IL-4 detection and performing ELISA. The antibody may be a great tool to study the immune responses in horses, particularly in scenarios of infections, immunological disorders or vaccinations.

Cite This Article

APA
Wagner B, Hillegas JM, Antczak DF. (2006). A monoclonal antibody to equine interleukin-4. Vet Immunol Immunopathol, 110(3-4), 363-367. https://doi.org/10.1016/j.vetimm.2006.01.001

Publication

Veterinary immunology and immunopathology
ISSN: 0165-2427
NlmUniqueID: 8002006
Country: Netherlands
Language: English
Volume: 110
Issue: 3-4
Pages: 363-367

Researcher Affiliations

Wagner, Bettina
  • Baker Institute for Animal Health, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853, USA. bw73@cornell.edu
Hillegas, Julie M
    Antczak, Douglas F

      MeSH Terms

      • Animals
      • Antibodies, Monoclonal / immunology
      • Cells, Cultured
      • Enzyme-Linked Immunosorbent Assay
      • Flow Cytometry
      • Horses / immunology
      • Interleukin-4 / immunology
      • Leukocytes, Mononuclear / immunology
      • Mice
      • Recombinant Proteins

      Citations

      This article has been cited 14 times.
      1. Keller LE, Tait Wojno ED, Begum L, Fortier LA. Interleukin-6 neutralization and regulatory T cells are additive in chondroprotection from IL-1β-induced inflammation. J Orthop Res 2023 May;41(5):942-950.
        doi: 10.1002/jor.25453pubmed: 36205183google scholar: lookup
      2. Gressler AE, Lübke S, Wagner B, Arnold C, Lohmann KL, Schnabel CL. Comprehensive Flow Cytometric Characterization of Bronchoalveolar Lavage Cells Indicates Comparable Phenotypes Between Asthmatic and Healthy Horses But Functional Lymphocyte Differences. Front Immunol 2022;13:896255.
        doi: 10.3389/fimmu.2022.896255pubmed: 35874777google scholar: lookup
      3. Schnabel CL, Fletemeyer B, Lübke S, Marti E, Wagner B, Alber G. CD154 Expression Indicates T Cell Activation Following Tetanus Toxoid Vaccination of Horses. Front Immunol 2022;13:805026.
        doi: 10.3389/fimmu.2022.805026pubmed: 35493462google scholar: lookup
      4. 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.
        doi: 10.1371/journal.pone.0252243pubmed: 34038479google scholar: lookup
      5. 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.
        doi: 10.4049/jimmunol.2001354pubmed: 33952617google scholar: lookup
      6. 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.
        doi: 10.1016/j.vetimm.2012.04.003pubmed: 22551980google scholar: lookup
      7. 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.
        doi: 10.1016/j.vetimm.2012.02.006pubmed: 22424928google scholar: lookup
      8. Wagner B, Burton A, Ainsworth D. Interferon-gamma, interleukin-4 and interleukin-10 production by T helper cells reveals intact Th1 and regulatory TR1 cell activation and a delay of the Th2 cell response in equine neonates and foals. Vet Res 2010 Jul-Aug;41(4):47.
        doi: 10.1051/vetres/2010019pubmed: 20374696google scholar: lookup
      9. de Mestre A, Noronha L, Wagner B, Antczak DF. Split immunological tolerance to trophoblast. Int J Dev Biol 2010;54(2-3):445-55.
        doi: 10.1387/ijdb.082795adpubmed: 19876828google scholar: lookup
      10. Butt SL, de Oliveira PSB, Rani R, Nooruzzaman M, Diaz AN, Glover S, Young AJ, Sharma B, Diel DG. Novel recombinant H5-based vaccine provides effective protection against H5N1 influenza virus in cats. NPJ Vaccines 2026 Jan 12;11(1):49.
        doi: 10.1038/s41541-025-01369-6pubmed: 41526385google scholar: lookup
      11. Wjst VF, Lübke S, Wagner B, Rhyner C, Jentsch MC, Arnold C, Lohmann KL, Schnabel CL. Aspergillus fumigatus antigen-reactive Th17 cells are enriched in bronchoalveolar lavage fluid in severe equine asthma. Front Immunol 2024;15:1367971.
        doi: 10.3389/fimmu.2024.1367971pubmed: 39229267google scholar: lookup
      12. Brooks MB, Brooks JC, Catalfamo J, Zhu Y, Goggs R, Babasyan S, Wagner B, LeVine DN. Plasma concentration of thrombopoietin in dogs with immune thrombocytopenia. J Vet Intern Med 2024 Sep-Oct;38(5):2507-2517.
        doi: 10.1111/jvim.17152pubmed: 39143652google scholar: lookup
      13. Holmes CM, Babasyan S, Wagner B. Neonatal and maternal upregulation of antileukoproteinase in horses. Front Immunol 2024;15:1395030.
        doi: 10.3389/fimmu.2024.1395030pubmed: 38736885google scholar: lookup
      14. Wang Y, Yang H, Hu J, Jiang Y, Ma W, Gao S, Chen D. Preparation and application of fluorescent monoclonal antibodies recognizing goat CD4(+)CD25(+) regulatory T cells. Appl Microbiol Biotechnol 2024 May 8;108(1):327.
        doi: 10.1007/s00253-024-13115-4pubmed: 38717623google scholar: lookup
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