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Effect of extracellular ionic calcium and magnesium on opsonic and non-opsonic phagocytosis of Escherichia coli bovine blood polymorphonuclear leucocytes.

Hilde Dosogne, Christian Burvenich UGent and Max Paape UGent (1998) COMPARATIVE HAEMATOLOGY INTERNATIONAL. 8(2). p.82-86
abstract
The effects of extracellular Ca2+ and Mg2+ concentrations on opsonic and non-opsonic phagocytosis of Escherichia call by bovine polymorphonuclear leucocytes (PMN) isolated from blood were evaluated by flow cytometry. Eight cows were used as blood donors. The green fluorescence of blood PMN selectively gated in the forward scatter (FS)- side scatter (SS) dot plot after incubation with fluorescein isothiocyanate (FITC) - labelled E. coli was used to characterise phagocytosis. Parameters for phagocytosis were percentage fluorescent PMN (% phagocytosis) and mean fluorescence intensity (MFI). The fluorescence of adherent bacteria was quenched with trypan blue to distinguish between adherence and ingestion. Nonopsonic and opsonic phagocytosis were decreased in the absence of extracellular ionic Ca2+ and Mg2+ compared to physiological levels. Addition of 10 mM EGTA to the incubation medium was necessary to block all extracellular Ca2+ and resulted in a significant decrease of opsonic phagocytosis, with only 5% phagocytic PMN after quenching. Increasing Ca2+ concentrations resulted in a gradual increase in percentage opsonic and non-opsonic phagocytosis and in MFI for opsonic phagocytosis. Ionic calcium plays an important role in phagocytosis (attachment as well as ingestion) by bovine blood PMN in the presence of opsonins, whereas non-opsonic phagocytosis appeared to be less dependent on Ca2+. However, reduced serum or milk calcium levels in cows are unlikely to cause a substantial reduction of PMN phagocytosis in vivo.
Please use this url to cite or link to this publication:
author
organization
year
type
journalArticle (original)
publication status
published
subject
keyword
ZYMOSAN PARTICLES, ALPHA-SUBUNIT, NEUTROPHIL PHAGOCYTOSIS, PERIPARTURIENT DISORDERS, COMPLEMENT RECEPTOR TYPE-3, phagocytosis, coliform mastitis, flow cytometry, HYPOCALCEMIA, LEUKOCYTES, MASTITIS, ADHESION, COWS
journal title
COMPARATIVE HAEMATOLOGY INTERNATIONAL
Comp. Haematol. Int.
volume
8
issue
2
pages
82-86 pages
Web of Science type
Article
Web of Science id
000074965900005
ISSN
0938-7714
DOI
10.1007/BF02642496
language
English
UGent publication?
yes
classification
A1
copyright statement
I have transferred the copyright for this publication to the publisher
id
179185
handle
http://hdl.handle.net/1854/LU-179185
date created
2004-01-14 13:41:00
date last changed
2016-12-19 15:38:09
@article{179185,
  abstract     = {The effects of extracellular Ca2+ and Mg2+ concentrations on opsonic and non-opsonic phagocytosis of Escherichia call by bovine polymorphonuclear leucocytes (PMN) isolated from blood were evaluated by flow cytometry. Eight cows were used as blood donors. The green fluorescence of blood PMN selectively gated in the forward scatter (FS)- side scatter (SS) dot plot after incubation with fluorescein isothiocyanate (FITC) - labelled E. coli was used to characterise phagocytosis. Parameters for phagocytosis were percentage fluorescent PMN (\% phagocytosis) and mean fluorescence intensity (MFI). The fluorescence of adherent bacteria was quenched with trypan blue to distinguish between adherence and ingestion. Nonopsonic and opsonic phagocytosis were decreased in the absence of extracellular ionic Ca2+ and Mg2+ compared to physiological levels. Addition of 10 mM EGTA to the incubation medium was necessary to block all extracellular Ca2+ and resulted in a significant decrease of opsonic phagocytosis, with only 5\% phagocytic PMN after quenching. Increasing Ca2+ concentrations resulted in a gradual increase in percentage opsonic and non-opsonic phagocytosis and in MFI for opsonic phagocytosis. Ionic calcium plays an important role in phagocytosis (attachment as well as ingestion) by bovine blood PMN in the presence of opsonins, whereas non-opsonic phagocytosis appeared to be less dependent on Ca2+. However, reduced serum or milk calcium levels in cows are unlikely to cause a substantial reduction of PMN phagocytosis in vivo.},
  author       = {Dosogne, Hilde and Burvenich, Christian and Paape, Max},
  issn         = {0938-7714},
  journal      = {COMPARATIVE HAEMATOLOGY INTERNATIONAL},
  keyword      = {ZYMOSAN PARTICLES,ALPHA-SUBUNIT,NEUTROPHIL PHAGOCYTOSIS,PERIPARTURIENT DISORDERS,COMPLEMENT RECEPTOR TYPE-3,phagocytosis,coliform mastitis,flow cytometry,HYPOCALCEMIA,LEUKOCYTES,MASTITIS,ADHESION,COWS},
  language     = {eng},
  number       = {2},
  pages        = {82--86},
  title        = {Effect of extracellular ionic calcium and magnesium on opsonic and non-opsonic phagocytosis of Escherichia coli bovine blood polymorphonuclear leucocytes.},
  url          = {http://dx.doi.org/10.1007/BF02642496},
  volume       = {8},
  year         = {1998},
}

Chicago
Dosogne, Hilde, Christian Burvenich, and Max Paape. 1998. “Effect of Extracellular Ionic Calcium and Magnesium on Opsonic and Non-opsonic Phagocytosis of Escherichia Coli Bovine Blood Polymorphonuclear Leucocytes.” Comparative Haematology International 8 (2): 82–86.
APA
Dosogne, Hilde, Burvenich, C., & Paape, M. (1998). Effect of extracellular ionic calcium and magnesium on opsonic and non-opsonic phagocytosis of Escherichia coli bovine blood polymorphonuclear leucocytes. COMPARATIVE HAEMATOLOGY INTERNATIONAL, 8(2), 82–86.
Vancouver
1.
Dosogne H, Burvenich C, Paape M. Effect of extracellular ionic calcium and magnesium on opsonic and non-opsonic phagocytosis of Escherichia coli bovine blood polymorphonuclear leucocytes. COMPARATIVE HAEMATOLOGY INTERNATIONAL. 1998;8(2):82–6.
MLA
Dosogne, Hilde, Christian Burvenich, and Max Paape. “Effect of Extracellular Ionic Calcium and Magnesium on Opsonic and Non-opsonic Phagocytosis of Escherichia Coli Bovine Blood Polymorphonuclear Leucocytes.” COMPARATIVE HAEMATOLOGY INTERNATIONAL 8.2 (1998): 82–86. Print.