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Phagocytosis of opsonized fluorescent microspheres by equine polymorphonuclear leukocytes
1Institute for Medical Microbiology, Infectious and Epidemic Diseases, Veterinary Faculty, University of Munich, München, FRG.
Abstract:
Equine blood polymorphonuclear leukocytes (PMN) were isolated by buffy coat and hypotonic lysis of residual erythrocytes. A highly reproducible method is described for measuring the uptake of opsonized latex microspheres by equine PMN using flowcytometry. The use of cytochalasin D allowed for differentiation of ingested from attached particles. The kinetics of phagocytosis in vitro is shown for different experimental conditions. We developed an assay for evaluation of phagocytic capacity of PMN which allows the assessment of drugs for their influence on phagocytosis in vivo as well as in vitro.
Insights
Researchers developed a new flow cytometry assay to measure how well equine polymorphonuclear leukocytes (PMN) engulf particles. This method aids in evaluating drugs that affect phagocytic capacity in horses.
Area of Science:
- Veterinary Immunology
- Cellular Immunology
- Equine Health
Background:
- Polymorphonuclear leukocytes (PMN) are crucial immune cells in horses.
- Assessing phagocytic capacity is vital for understanding equine immune function.
- Existing methods for evaluating phagocytosis in equine PMN are limited.
Purpose of the Study:
- To develop and validate a reproducible flow cytometry assay for equine PMN phagocytosis.
- To differentiate between ingested and attached particles using cytochalasin D.
- To assess the phagocytic capacity of equine PMN under various experimental conditions.
Main Methods:
- Isolation of equine PMN from buffy coat with hypotonic lysis.
- Quantification of opsonized latex microsphere uptake via flow cytometry.
- Use of cytochalasin D to distinguish ingested from surface-adhered particles.
Main Results:
- A highly reproducible flow cytometry method for measuring equine PMN phagocytosis was established.
- The assay effectively differentiates ingested particles from attached ones.
- Phagocytosis kinetics were characterized under different in vitro conditions.
Conclusions:
- The developed assay provides a reliable tool for evaluating equine PMN phagocytic capacity.
- This method facilitates the assessment of drug effects on phagocytosis, both in vitro and in vivo.
- The assay supports research into equine immune responses and potential therapeutic interventions.