Phagocytic response of bovine polymorphonuclear leukocytes to different incubation conditions and following exposure to some effectors of phagocytosis and different anticoagulants in vitro

The ability of bovine polymorphonuclear leukocytes (PMN) to phagocytose fluorescent beads in vitro was studied using flow cytometry. The effects of varying laboratory conditions (bead:PMN ratio, length of incubation, and temperature) were first determined, then the effects of lipopolysaccharide (LPS...

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Bibliographic Details
Published in:Canadian journal of veterinary research 2001, Vol.65 (1), p.38-44
Main Authors: Ducusin, R.J.T, Sarashina, T, Uzuka, Y, Tanabe, S, Ohtani, M
Format: Article
Language:English
Subjects:
Animals
anticoagulants
Anticoagulants - pharmacology
Cattle - blood
Cattle - immunology
cell culture
cytochalasin B
Cytochalasin B - pharmacology
dairy cows
dosage
dose response
drug effects
Female
flow cytometry
Flow Cytometry - veterinary
formyl-met-leu-phe
lipopolysaccharides
Lipopolysaccharides - pharmacology
myristic acid
neutrophils
Neutrophils - immunology
oligopeptides
phagocytosis
Phagocytosis - drug effects
phorbol myristate acetate
Temperature
Tetradecanoylphorbol Acetate - pharmacology
Time Factors
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Summary:The ability of bovine polymorphonuclear leukocytes (PMN) to phagocytose fluorescent beads in vitro was studied using flow cytometry. The effects of varying laboratory conditions (bead:PMN ratio, length of incubation, and temperature) were first determined, then the effects of lipopolysaccharide (LPS), phorbol myristate acetate (PMA), cytochalasin B, and formyl-met-leu-phe (fMLP) on phagocytosis were evaluated. The recommended bead:PMN ratio, incubation period, and incubation temperature are 20:1, 30 min, and 38.5 degrees C, respectively. Lipopolysaccharide increased phagocytosis at a relatively high minimum dose; PMA increased phagocytosis even at low doses; cytochalasin B increased and decreased phagocytosis at low and high doses, respectively; and fMLP had no significant effect on phagocytosis. Also, the effects of ethylene diamine tetraacetic acid (EDTA) and acid citrate dextrose (ACD) as anticoagulants were compared with heparin-treated blood PMNs. Both EDTA and ACD decreased phagocytosis. Although there are reports that demonstrated that heparin reduced PMN phagocytosis, at least among the 3 anticoagulants used, heparin remains to be the standard anticoagulant for the study of PMN phagocytosis.
ISSN:0830-9000