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Home / Equine Research Bank / Immunology / Three monoclonal antibodies identifying antigens on all equi...
Immunology1991; 74(2); 251-257;

Three monoclonal antibodies identifying antigens on all equine T lymphocytes, and two mutually exclusive T-lymphocyte subsets.

Abstract: The aim of this study was to produce monoclonal antibodies (mAb) recognizing equine lymphocyte surface antigens. Fusions were conducted using BALB/c mice hyperimmunized with equine thymocytes. Hybridoma supernatants were screened by flow cytometry and positive hybridomas were cloned twice by limiting dilution. These mAb were then characterized for tissue distribution by immunohistology and flow cytometry, and by precipitation and analysis of the lymphocyte antigens which they recognized. Three mAb (CVS5, CVS4 and CVS8) are described which recognize only T lymphocytes in peripheral blood. Two-colour immunofluorescent studies showed that CVS5 recognized all T lymphocytes and that CVS4 and CVS8 recognized two mutually exclusive subsets of CVS5-positive cells. In the thymus there was a large population of CVS4/CVS8 double-positive cells. Immunohistochemical staining with these mAb was restricted to T-lymphocyte areas. CVS4 and CVS5 precipitated molecules of 58,000 and 69,000 MW, respectively, in both reducing and non-reducing conditions. CVS8 precipitated two molecules of 32,000 and 39,000 MW in reducing conditions, and one molecule of 69,000 MW in non-reducing conditions. This evidence suggests that CVS5, CVS4 and CVS8 recognize the equine homologues of CD5, CD4 and CD8, and that the characteristics of these antigens are similar to those of other species.
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Publication Date: 1991-10-01 PubMed ID: 1748472PubMed Central: PMC1384601
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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 study focuses on generating monoclonal antibodies that can identify specific antigens on all equine (horse) T lymphocytes, as well as two distinctive T-lymphocyte subsets. The investigation highlighted the possibility of the monoclonal antibodies recognizing homologues of CD5, CD4 and CD8 in horses, which are common in other species.

Monoclonal Antibody Production

  • The researchers began by producing monoclonal antibodies (mAb) designed to identify equine lymphocyte surface antigens. The process required the use of BALB/c mice that were hyperimmunized with equine thymocytes.
  • Hybridoma supernatants, the cells that produce mAb, were examined by flow cytometry to identify any positive hybridoma clones that were then cloned twice using the limiting dilution method to ensure purity of antibodies.

Antibody Characterization

  • Characterization involved studying tissue distribution through immunohistology and flow cytometry, along with identifying the specific lymphocyte antigens the mAb could recognize through precipitation and analysis.
  • The study yielded three mAb (CVS5, CVS4 and CVS8) that were found to only recognize T lymphocytes floating in the peripheral blood system.

T-Lymphocyte Subsets Identification

  • Immunofluorescent studies using two colors showed that the CVS5 mAb recognized all T lymphocytes, while CVS4 and CVS8 identified two separate subsets of CVS5-positive cells that did not overlap.
  • The thymus of the subject animals showed that there was an abundance of T cells that could be labeled by both CVS4 and CVS8, indicating the evidence of two separate subsets of T-cells.
  • Immunohistochemical staining using these mAb was restricted only to areas where T lymphocytes were found.

Molecule Precipitation

  • During the precipitation process, CVS4 and CVS5 precipitated molecules with a molecular weight of 58,000 and 69,000, respectively, under both reducing and non-reducing conditions.
  • CVS8 precipitated two separate molecules weighing 32,000 and 39,000 under reducing conditions, and just a single molecule weighing 69,000 under non-reducing conditions.
  • The properties of these molecules suggest that they are the equine homologues equivalent to CD5, CD4 and CD8 in other species.

Cite This Article

APA
Lunn DP, Holmes MA, Duffus WP. (1991). Three monoclonal antibodies identifying antigens on all equine T lymphocytes, and two mutually exclusive T-lymphocyte subsets. Immunology, 74(2), 251-257.

Publication

Immunology
ISSN: 0019-2805
NlmUniqueID: 0374672
Country: England
Language: English
Volume: 74
Issue: 2
Pages: 251-257

Researcher Affiliations

Lunn, D P
  • Department of Clinical Veterinary Medicine, University of Cambridge, U.K.
Holmes, M A
    Duffus, W P

      MeSH Terms

      • Animals
      • Antibodies, Monoclonal / biosynthesis
      • Antigens, Surface / analysis
      • Antigens, Surface / chemistry
      • Cell Separation / methods
      • Flow Cytometry
      • Horses / immunology
      • Immunoenzyme Techniques
      • Molecular Weight
      • T-Lymphocyte Subsets / immunology
      • T-Lymphocytes / immunology
      • Thymus Gland / immunology

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      This article has been cited 9 times.
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