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Flow-cytometric Studies of the Phagocytic Capacities of Equine Neutrophils

Flödescytometriska studier av fagocytosförmågan hos ekvina neutrofiler

Acta Veterinaria Scandinavica volume 36, pages 553–562 (1995)Cite this article

Abstract

Methodological aspects of flow-cytometric evaluation of the phagocytic properties of equine neutrophils were elucidated. The kinetics of attachment and ingestion were studied, and the phagocytic process was more rapidly completed when serum-opsonized yeast cells were used than with use of IgG-opsonized yeast cells. Trypan blue was successfully used to quench fluorescence of non-ingested yeast cells. There were only minor differences in the kinetics of phagocytosis between quenched and un-quenched samples, indicating that attachment is rapidly followed by ingestion. Trypan blue quenching caused loss of cells with light scattering properties of granulocytes, although this did not affect the determined frequencies of truly phagocytic neutrophils. Aggregation of yeast cells proved to be a disturbance but not an obstacle to the determination of frequencies of actively phagocytic cells. Flow cytometry is well suited for studies of phagocytosis of yeast cells by equine neutrophils, and the trypan blue quenching provides a means of eliminating false-positive events due to aggregation of yeast cells. The main advantage of the flow-cytometric method is the possibility of rapid processing of a large number of samples, making the method useful for studies of herds.

Sammanfattning

Den fagocyterande förmågan hos hästens neutrofiler studerades med flödescytometri. Kinetiken för adhesion och upptag av jästceller undersöktes. Fagocytosförloppet slutfördes snabbare när serum-opsoniserade jästceller användes än när IgG-opsomgerade jästceller var målpartiklar. Släckning med trypanblått ger möjlighet att eliminera falskt positiva händelser, som beror på jästaggregat utanför neutrofilen. Släckningen orsakade en förlust av celler med neutrofilers ljus spridningsegenskaper, men detta påverkade inte de uppmätta frekvenserna av fagocylerande neutrofiler. Endast smärre skillnader i fagocytoskinetiken kunde ses när släckta och osläckta prover jämfördes. Det konkluderas att flödes-cytometri är väl lämpat för studier av ekvina neutrofilers fagocytos av jästceller. Metodens största fördel är att man snabbt kan analysera stora provserier, vilket möjliggör undersökningar av hela besättningar.

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Acknowledgments

The financial support from the Swedish Horse Race Totalizator Board (ATG) is gratefully acknowledged. The flow cytometer used in this study was financed by a grant from the Knut and Alice Wallenberg foundation.

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Authors and Affiliations

  1. Department of Pathology, Faculty of Veterinary Medicine, Swedish University of Agricultural Sciences, P.O. Box 7028, S-750 07, Uppsala, Sweden

    A. Johannisson

  2. Department of Medicine and Surgery, Faculty of Veterinary Medicine, Swedish University of Agricultural Sciences, Uppsala, Sweden

    G. Gröndahl, S. Demmers & M Jensen-Waern

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  1. A. Johannisson
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  2. G. Gröndahl
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  3. S. Demmers
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  4. M Jensen-Waern
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Johannisson, A., Gröndahl, G., Demmers, S. et al. Flow-cytometric Studies of the Phagocytic Capacities of Equine Neutrophils. Acta Vet Scand 36, 553–562 (1995). https://doi.org/10.1186/BF03547669

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Acta Veterinaria Scandinavica

ISSN: 1751-0147