Production and characterization of a monoclonal antibody recognizing a cytoplasmic antigen of equine mononuclear phagocytes.
Abstract: An IgG1 mouse monoclonal antibody, designated 1.646, is described which recognizes a cytoplasmic antigen of equine mononuclear phagocytes. Indirect fluorescent antibody staining of peripheral blood leukocytes reveals a granular cytoplasmic staining, predominantly in adherent blood mononuclear cells. Indirect fluorescent antibody staining is positive for alveolar and peritoneal macrophages. In some horses, a few neutrophils are also stained. In equine tissue samples stained by immunohistochemistry, the distribution of positive cells is consistent with the distribution of tissue macrophages. The most intense and reliable staining occurs with splenic and lymph node macrophages. Hepatic Kupffer cells also stain with antibody 1.646, although the intensity of that staining is somewhat variable between horses. A granular pattern of staining typical of lipofuscin deposition is also seen in liver sections. There is also pale staining of some biliary and renal tubular epithelium. Equine erythrocytes, platelets and lymphocytes are not recognized by this antibody, and neither are monocyte/macrophages of human, canine or feline origin. Antibody 1.646 recognizes two proteins (150 and 30 kDa) of equine monocyte-derived macrophages when assayed by Western immunoblot. Because of the distribution of staining (tissue mononuclear phagocytes, lipofuscin-containing storage granules, biliary and renal tubular epithelium, and some neutrophils) we hypothesize that antibody 1.646 recognizes a cytoplasmic antigen that is closely associated with lysosomal membranes.
Publication Date: 1993-05-01 PubMed ID: 8333142DOI: 10.1016/0165-2427(93)90027-2Google Scholar: Lookup
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- Journal Article
- Research Support
- U.S. Gov't
- Non-P.H.S.
- Research Support
- U.S. Gov't
- P.H.S.
Summary
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The study details the production of a mouse monoclonal antibody, named 1.646, that successfully recognizes a specific antigen located in the cell component of horse mononuclear phagocytes. This capability was confirmed through fluorescence and observed through unique patterns in different tissue samples. It is suggested that the recognized antigen may be closely connected to lysosomal membranes.
Antibody 1.646 and its Antigen Recognition
- The research generates an IgG1 mouse monoclonal antibody, named 1.646 which identifies a cytoplasmic antigen found in equine mononuclear phagocytes.
- Through indirect fluorescent antibody staining of peripheral blood leukocytes, predominantly observable in adherent blood mononuclear cells, it is confirmed that the antibody successfully recognizes the antigen.
- this antibody-antigen interaction elicits a granular cytoplasmic staining.
Fluorescent Staining and Observations
- The antibody is revealed to positively stain alveolar and peritoneal macrophages through indirect fluorescent antibody staining methods.
- A small number of neutrophils from some horses also showed positive staining.
- Through immunohistochemistry of equine tissue samples, it is observed that the presence of cells displaying positive staining was consistent with the distribution of tissue macrophages.
- Macrophages in splenic and lymph node regions displayed the most intense and reliable staining patterns.
Distribution of Staining and Interpretations
- Hepatic Kupffer cells and variable intensities of liver sections also demonstrated staining with this antibody, indicating that the recognized antigen is present across different tissue types.
- The antibody also recognizes two proteins, of 150 and 30 kDa, in equine monocyte-derived macrophages, when subjected to Western immunoblot assay.
- However, the antibody did not recognize erythrocytes, platelets, lymphocytes from horses, or monocytes/macrophages from humans, dogs and cats, suggesting that the antigen is exclusive to equine mononuclear phagocytes.
- Based on the distribution of staining and antibody-antigen interactions observed, it is hypothesized that antibody 1.646 recognizes a cytoplasmic antigen closely associated with lysosomal membranes.
Cite This Article
APA
Sellon DC, Cullen JM, Whetter LE, Gebhard DH, Coggins L, Fuller FJ.
(1993).
Production and characterization of a monoclonal antibody recognizing a cytoplasmic antigen of equine mononuclear phagocytes.
Vet Immunol Immunopathol, 36(4), 303-318.
https://doi.org/10.1016/0165-2427(93)90027-2 Publication
Researcher Affiliations
- Department of Microbiology, Pathology and Parasitology, North Carolina State University College of Veterinary Medicine, Raleigh 27606.
MeSH Terms
- Animals
- Antibodies, Monoclonal / biosynthesis
- Antibodies, Monoclonal / immunology
- Antibodies, Viral / biosynthesis
- Antibody Specificity
- Antigens, Viral / immunology
- Blotting, Western / veterinary
- Equine Infectious Anemia / etiology
- Equine Infectious Anemia / immunology
- Fluorescent Antibody Technique / veterinary
- Horses / immunology
- Hybridomas
- Immunoglobulin G / biosynthesis
- Immunoglobulin G / immunology
- Leukocytes, Mononuclear / immunology
- Liver / immunology
- Lung / immunology
- Lymph Nodes / immunology
- Macrophages / immunology
- Mice
- Phagocytes / immunology
- Spleen / immunology
Grant Funding
- 1K11-AI00963 / NIAID NIH HHS
- R01-AI24904 / NIAID NIH HHS
Citations
This article has been cited 1 times.- Sellon DC, Walker KM, Russell KE, Perry ST, Covington P, Fuller FJ. Equine infectious anemia virus replication is upregulated during differentiation of blood monocytes from acutely infected horses. J Virol 1996 Jan;70(1):590-4.
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