A new surface marker on equine peripheral blood lymphocytes. I. Subpopulations of lymphocytes with receptors for Helix pomatia A hemagglutinin (HP).
Abstract: Untreated and neuraminidase-treated equine peripheral blood lymphocytes were analysed for binding of the A hemagglutinin of the snail Helix pomatia (HP). For optimal staining by direct immunofluorescence, the concentration of neuraminidase had to be increased as compared to that needed for other species. Moreover, higher concentrations of HP were required for optimal staining of equine lymphocytes as compared to lymphocytes from other species. Even so, the maximal number of equine lymphocytes exhibiting positive staining was only about 20%. No, or very few, HP-positive lymphocytes were seen when neuraminidase treatment was omitted. However, when the more sensitive method of indirect immunofluorescence was used, approximately 60% of the lymphocytes were HP positive without prior treatment with this enzyme. Neuraminidase treatment significantly increased this figure to about 75%. In all instances, HP binding was specific since it was inhibited by the competitive sugar hapten N-acetyl-D-galactosamine (D-GalNAc) while addition of D-glucose (D-Glc) gave no inhibition. HP binding to neuraminidase-treated lymphocytes was also investigated quantitatively by means of 125I-labeled HP. The number of HP molecules bound per HP-positive cell was approximately 3 X 10(5) and the apparent association constant for the binding of HP to its cellular receptors was approximately 8 X 10(7) 1/mol. No binding of HP to untreated lymphocytes could be recorded in these experiments.
Publication Date: 1985-01-01 PubMed ID: 3976169DOI: 10.1016/0165-2427(85)90108-4Google Scholar: Lookup
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- Comparative Study
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research carries out a study on equine peripheral blood lymphocytes and their affinity for Helix pomatia A hemagglutinin (HP), a compound found in a specific type of snail. Notably, to achieve maximal staining in the immunofluorescence application, higher concentrations of neuraminidase and HP were required compared to other species.
Subject of the Research
- The study revolves around equine peripheral blood lymphocytes, which are white blood cells that play essential roles in the body’s immune system.
- The research investigates the lymphocytes’ interaction with the A hemagglutinin of the snail Helix pomatia (HP), a complex that causes the clumping of cells, especially red blood cells.
Findings of the Research
- The study elucidates that treating equine lymphocytes with neuraminidase, an enzyme involved in breaking down glycosidic bonds, significantly enhances the binding capability of these cells to HP. The optimal staining, by direct immunofluorescence, required this neuraminidase treatment and higher concentrations of HP compared to other species.
- Despite these extensive treatments, only about 20% of equine lymphocytes showed positive staining for HP binding.
- The experiment also discovered that in contrast to the direct method, using indirect immunofluorescence increased the percentage of HP-positive lymphocytes to about 60% even without neuraminidase treatment – and this figure rose to about 75% when neuraminidase treatment was used.
Specificity of HP Binding
- The specificity of HP binding was confirmed as its interaction was inhibited by the competitive sugar hapten N-acetyl-D-galactosamine (D-GalNAc), while the addition of D-glucose (D-Glc) had no inhibitory effect.
- Quantitative analysis was performed using radiolabeled HP (125I-HP) showed an apparent association constant and the number of HP molecules bound per HP-positive cell. This provided critical data regarding the binding affinity and the extent of HP-cellular interaction.
Conclusion
- In conclusion, the research article emphasizes the detailed analysis of HP binding with equine peripheral blood lymphocytes and the significant role that neuraminidase plays in facilitating this binding.
Cite This Article
APA
Broström H, Hellström U, Hammarström S, Obel N, Perlmann P.
(1985).
A new surface marker on equine peripheral blood lymphocytes. I. Subpopulations of lymphocytes with receptors for Helix pomatia A hemagglutinin (HP).
Vet Immunol Immunopathol, 8(1-2), 35-46.
https://doi.org/10.1016/0165-2427(85)90108-4 Publication
Researcher Affiliations
MeSH Terms
- Animals
- Female
- Horses / blood
- Horses / immunology
- Kinetics
- Lectins / immunology
- Lymphocytes / classification
- Lymphocytes / immunology
- Male
- Neuraminidase
- Receptors, Mitogen
Citations
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