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Flow Cytometric Characterization of Bovine Blood Neutrophil Phagocytosis of Fluorescent Bacteria and Zymosan Particles

Flödescytometrisk karaktärisering av bovina neutrofila leukocyters fagocytos av fluorescerande bakterier och zymosan partiklar

Acta Veterinaria Scandinavica volume 26, pages 289–307 (1985)Cite this article

Abstract

A Flow Cytometric method for the evaluation of the phagocytic capacity of bovine blood neutrophils is described. The neutrophils were isolated from bovine blood by a one step discontinuous gradient of Percoll. By this technique of isolation, 90 ± 2.8 % (mean ± s) of the granulocytes in the whole blood were recovered.

Isolated neutrophils were incubated with FITC labeled S. aureus or zymosan particles in a ratio of 1:20 and 1:10, respectively, and a final serum concentration of 10 %. Phagocytosis was terminated after 15 min and the number of extracellular bacteria or zymosan particles and the percentage of phagocytic granulocytes were registered by Flow Cytometry (FCM). FCM and microscopic studies revealed that eosinophils play a minor role in the phagocytosis of bacteria. The neutrophils were the main population of the granulocytes which were actively phagocytic. Variation among cows in the ability of their blood neutrophils to phagocytize bacteria was evident.

Sammanfattning

En flödescytometrisk metod användes för att studera fagocytos-kapaciteten hos neutrofila leukocyter som isolerades från bovint blod. För isolering av cellerna från blodet användes en enstegs diskontinuerlig Percoll gradient. Med metoden kunde 90 ± 2,8 % (medeltal ± SD) av granulocyterna utvinnas ur blodet.

De neutrofila leukocyterna inkuberades med FITC (fluorecin isothiocyanat) märkta S. aureus eller zymosanpartiklar i proportionerna 1:20 respektive 1:10 i 10 % serum. Fagocytosen avslutades efter 15 minut er och antatet extracellulära bakterier eller zymosanpartiklar och fagocyterande granulocyter mättes medelst flödescytometri (FGM). FGM och mikroskopiska studier visade att de eosinofila leukocyterna fagocyterar bakterier i ringa utsträckning. Den granulocytpopulation som var aktivt fagocyterande utgjordes till allra största delen av neutrofila luekocyter.

De neutrofila leukocyternas förmåga at fagocytera bakterier varierade melian kor.

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Authors and Affiliations

  1. Department of Obstetrics and Gynaecology and Department of Animal Breeding and Genetics, Swedish University of Agricultural Sciences, S-750 07, Uppsala 7, Sweden

    Alaa M. Saad & M. Hageltorn

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  1. Alaa M. Saad
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  2. M. Hageltorn
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Saad, A.M., Hageltorn, M. Flow Cytometric Characterization of Bovine Blood Neutrophil Phagocytosis of Fluorescent Bacteria and Zymosan Particles. Acta Vet Scand 26, 289–307 (1985). https://doi.org/10.1186/BF03546530

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Acta Veterinaria Scandinavica

ISSN: 1751-0147