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 language | English (US) |
|---|---|
| Pages (from-to) | 47-55 |
| Number of pages | 9 |
| Journal | Comparative Haematology International |
| Volume | 5 |
| Issue number | 1 |
| DOIs | |
| State | Published - Mar 1995 |
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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
Cite this
Modulation of phagocytic and oxidative burst activities of bovine neutrophils by human recombinant TNF-α, IL-1-α, IFN-γ, G-CSF, GM-CSF. / Kabbur, M. B.; Jain, N. C.; Farver, Thomas B.
In: Comparative Haematology International, Vol. 5, No. 1, 03.1995, p. 47-55.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Modulation of phagocytic and oxidative burst activities of bovine neutrophils by human recombinant TNF-α, IL-1-α, IFN-γ, G-CSF, GM-CSF
AU - Kabbur, M. B.
AU - Jain, N. C.
AU - Farver, Thomas B
PY - 1995/3
Y1 - 1995/3
N2 - 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.
AB - 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.
KW - Bovine
KW - Cytokines
KW - Granulocyte colonystimulating factor
KW - Granulocyte-macrophage colonystimulating factor
KW - Hydrogen peroxide production
KW - Interferon-gamma
KW - Interleukin-1 alpha
KW - Neutrophils
KW - Phagocytosis
KW - Tumour necrosis factor-alpha
UR - http://www.scopus.com/inward/record.url?scp=0029073680&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0029073680&partnerID=8YFLogxK
U2 - 10.1007/BF00214490
DO - 10.1007/BF00214490
M3 - Article
AN - SCOPUS:0029073680
VL - 5
SP - 47
EP - 55
JO - Comparative Clinical Pathology
JF - Comparative Clinical Pathology
SN - 1618-5641
IS - 1
ER -