Modulation of phagocytic and oxidative burst activities of bovine neutrophils by human recombinant TNF-α, IL-1-α, IFN-γ, G-CSF, GM-CSF

M. B. Kabbur, N. C. Jain, Thomas B Farver

Research output: Contribution to journalArticle

17 Scopus citations

Abstract

In vitro experiments were conducted to determine the effects of recombinant cytokines on phagocytic and oxidative burst activities of bovine neutrophils. Neutrophils were isolated (purity >91%, viability >97%) from EDTA-anticoagulated blood from healthy Holstein-Friesian heifers. Aliquots of neutrophils (10 × 106 cells/ml) were incubated for 1 h at 37 °C with equal volumes of recombinant human cytokines, namely, tumour necrosis factor-alpha (rhTNF-α, 0.5-1000 ng/ml), interleukin-1-alpha (rhIL-1-α, 0.001-10 ng/ml), interferon-gamma (rhIFN-γ, 0.01-100 ng/ml), granulocyte colony-stimulating factor (rhG-CSF, 25 ng/ml), and granulocyte-macrophage colony-stimulating factor (rhGM-CSF, 10 ng/ml). Then, the percentage phagocytosis and average number of intracellular bacteria per cell were evaluated by flow cytometry and/or fluorescent microscopy using FITC-labelled opsonised bacteria (Escherichia coli 0111:B4). Unlabelled opsonised bacteria and dichlorofluorescin diacetate were used to evaluate H2O2 production, a measure of oxidative burst, by flow cytometry. The results showed that all five cytokines significantly (p <0.05) increased percentage phagocytosis (52.4-86.1%), number of intracellular bacteria per cell (24.9-47.9%), and H2O2 production (31.3-58.2%) when compared to untreated neutrophils. A gradual increase in mean channel fluorescence but not in percentage phagocytosis was consistently seen with increasing concentrations of rhTNF-α, rhIL-1-α, and rhIFN-γ, thereby indicating a concentration-dependent stimulation of phagocytic capacity by these three cytokines.

Original languageEnglish (US)
Pages (from-to)47-55
Number of pages9
JournalComparative Haematology International
Volume5
Issue number1
DOIs
StatePublished - Mar 1995

Keywords

  • Bovine
  • Cytokines
  • Granulocyte colonystimulating factor
  • Granulocyte-macrophage colonystimulating factor
  • Hydrogen peroxide production
  • Interferon-gamma
  • Interleukin-1 alpha
  • Neutrophils
  • Phagocytosis
  • Tumour necrosis factor-alpha

ASJC Scopus subject areas

  • Hematology

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