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A Simple Fluorescence Assay for Quantification of Canine Neutrophil Extracellular Trap Release
Published on: November 21, 2016
Evaluation in vitro of canine neutrophil function
S Comazzi1, S Paltrinieri, V Spagnolo
1Dipartimento di Patologia Animale, Igiene e Sanità Pubblica Veterinaria, Sezione di Patologia Generale Veterinaria e Parassitologia, via Celoria 10, 20133 Milano, Italy. stex@mailserver.unimi.it
Veterinary Journal (London, England : 1997)
|October 30, 2001
Summary
This study established normal values for canine polymorphonuclear granulocyte (PMN) phagocytosis tests. Standardized methods were developed for assessing PMN function in dogs, crucial for diagnosing various diseases.
Area of Science:
- Veterinary Immunology
- Canine Hematology
- Cellular Immunology
Background:
- Polymorphonuclear granulocyte (PMN) phagocytosis is vital for immune response.
- Pathological changes can significantly impact PMN function.
- Establishing normal canine PMN phagocytosis values is essential for disease diagnosis.
Purpose of the Study:
- To obtain normal values for canine PMN phagocytosis assays.
- To standardize techniques for evaluating PMN function in dogs.
- To assess various aspects of phagocytosis, including chemotaxis, yeast uptake, superoxide production, and adherence.
Main Methods:
- Isolation of PMNs using discontinuous Percoll gradients.
- Chemotaxis assessed via modified Boyden chamber (leading front method).
- Fluorescinated yeast uptake evaluated on a slide.
- Superoxide (SO) production and adherence measured using microtitre plates.
- Optimized incubation periods and activators (interleukin-8 for chemotaxis, phorbol myristate acetate for SO production) were determined.
Main Results:
- Normal values for canine PMN phagocytosis assays were established.
- Standardized protocols for PMN isolation and functional testing were developed.
- No significant correlation was found between different phagocytic functions tested.
- Optimized incubation times (60 min for chemotaxis, 30 min for SO production) yielded better results.
Conclusions:
- The study successfully standardized and obtained normal values for key canine PMN phagocytosis assays.
- These standardized methods and values are crucial for accurate interpretation of PMN function in clinical veterinary diagnostics.
- The lack of correlation between different phagocytic aspects suggests they measure distinct functional capabilities.
