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Evaluation of L-selectin expression and assessment of protein tyrosine phosphorylation in bovine polymorphonuclear neutrophil leukocytes around parturition.

Erica Monfardini, Max Paape UGent, Yan WANG, Anthony Capuco UGent, Michael HUSHEEM, Larry WOOD and Christian Burvenich UGent (2002) VETERINARY RESEARCH. 33(3). p.271-281
abstract
Impaired polymorphonuclear neutrophil leukocyte (PMN) function around parturition has been associated with increased clinical mastitis in dairy cows. Rolling and attachment of PMN to the endothelium is the first step in the recruitment process and is accomplished by interaction between L-selectin on PMN and its ligand on endothelial cells. Furthermore, tyrosine phosphorylation is involved in the initiation of many PMN functions. The objective of this work was to determine changes in expression of L-selectin and tyrosine phosphorylation in the perinatal period. Eight clinically healthy Holstein cows were used as PMN donors at d-21, -14, -7,0 (calving), +1, +2, +7, +14, +28. Evaluation of L-selectin expression was carried out on activated and resting PMN. Anti-bovine L-selectin monoclonal antibody (MAB) and flow cytometric analysis were used to measure the percentage of PMN fluorescing and receptor expression (log mean fluorescent channel, LMFC). Activated and resting PMN showed similar trends in % PMN fluorescence and LMFC. The percentage of PMN fluorescing tended to decrease at parturition, followed by a significant increase at d +14 and +28 (P <0.02). For LMFC a decrease was observed on d +1 followed by an increase through d +28 (P < 0.01). Protein tyrosine phosphorylation of lysates prepared from PMN isolated throughout the study was detected by electrophoresis and western blotting using anti-phosphotyrosine MAB. Several protein bands were tyrosine phosphorylated. Two of these bands (42-44 kDa and 90 kDa) varied in intensity over time. The intensity of the 42-44 kDa band gradually increased from d -7, peaked at d +7 (P < 0.03), and steadily decreased to d +28 (P < 0.02). Antibody to activated mitogen protein kinase reacted with the 42-44 kDa band. Reduced PMN function during the periparturient period could be related to reduced L-selectin adhesion molecules on the cell surface, and to modulation in the phosphorylation of functionally important molecules.
Please use this url to cite or link to this publication:
author
organization
year
type
journalArticle (original)
publication status
published
subject
keyword
BLOOD, MILK, DAIRY-COWS, MAP KINASE, SIGNAL-TRANSDUCTION, ADHESION MOLECULES, parturition, mastitis, neutrophil, L-selectin, tyrosine phosphorylation, DEXAMETHASONE, PHAGOCYTOSIS, AGGREGATION, RECRUITMENT
journal title
VETERINARY RESEARCH
Vet. Res.
volume
33
issue
3
pages
271-281 pages
Web of Science type
Article
Web of Science id
000175765600005
JCR category
VETERINARY SCIENCES
JCR impact factor
2.031 (2002)
JCR rank
4/129 (2002)
JCR quartile
1 (2002)
ISSN
0928-4249
DOI
10.1051/vetres:2002015
language
English
UGent publication?
yes
classification
A1
copyright statement
I have transferred the copyright for this publication to the publisher
id
149257
handle
http://hdl.handle.net/1854/LU-149257
date created
2004-01-14 13:38:00
date last changed
2016-12-19 15:37:50
@article{149257,
  abstract     = {Impaired polymorphonuclear neutrophil leukocyte (PMN) function around parturition has been associated with increased clinical mastitis in dairy cows. Rolling and attachment of PMN to the endothelium is the first step in the recruitment process and is accomplished by interaction between L-selectin on PMN and its ligand on endothelial cells. Furthermore, tyrosine phosphorylation is involved in the initiation of many PMN functions. The objective of this work was to determine changes in expression of L-selectin and tyrosine phosphorylation in the perinatal period. Eight clinically healthy Holstein cows were used as PMN donors at d-21, -14, -7,0 (calving), +1, +2, +7, +14, +28. Evaluation of L-selectin expression was carried out on activated and resting PMN. Anti-bovine L-selectin monoclonal antibody (MAB) and flow cytometric analysis were used to measure the percentage of PMN fluorescing and receptor expression (log mean fluorescent channel, LMFC). Activated and resting PMN showed similar trends in \% PMN fluorescence and LMFC. The percentage of PMN fluorescing tended to decrease at parturition, followed by a significant increase at d +14 and +28 (P {\textlangle}0.02). For LMFC a decrease was observed on d +1 followed by an increase through d +28 (P {\textlangle} 0.01). Protein tyrosine phosphorylation of lysates prepared from PMN isolated throughout the study was detected by electrophoresis and western blotting using anti-phosphotyrosine MAB. Several protein bands were tyrosine phosphorylated. Two of these bands (42-44 kDa and 90 kDa) varied in intensity over time. The intensity of the 42-44 kDa band gradually increased from d -7, peaked at d +7 (P {\textlangle} 0.03), and steadily decreased to d +28 (P {\textlangle} 0.02). Antibody to activated mitogen protein kinase reacted with the 42-44 kDa band. Reduced PMN function during the periparturient period could be related to reduced L-selectin adhesion molecules on the cell surface, and to modulation in the phosphorylation of functionally important molecules.},
  author       = {Monfardini, Erica and Paape, Max and WANG, Yan and Capuco, Anthony and HUSHEEM, Michael and WOOD, Larry and Burvenich, Christian},
  issn         = {0928-4249},
  journal      = {VETERINARY RESEARCH},
  keyword      = {BLOOD,MILK,DAIRY-COWS,MAP KINASE,SIGNAL-TRANSDUCTION,ADHESION MOLECULES,parturition,mastitis,neutrophil,L-selectin,tyrosine phosphorylation,DEXAMETHASONE,PHAGOCYTOSIS,AGGREGATION,RECRUITMENT},
  language     = {eng},
  number       = {3},
  pages        = {271--281},
  title        = {Evaluation of L-selectin expression and assessment of protein tyrosine phosphorylation in bovine polymorphonuclear neutrophil leukocytes around parturition.},
  url          = {http://dx.doi.org/10.1051/vetres:2002015},
  volume       = {33},
  year         = {2002},
}

Chicago
Monfardini, Erica, Max Paape, Yan WANG, Anthony Capuco, Michael HUSHEEM, Larry WOOD, and Christian Burvenich. 2002. “Evaluation of L-selectin Expression and Assessment of Protein Tyrosine Phosphorylation in Bovine Polymorphonuclear Neutrophil Leukocytes Around Parturition.” Veterinary Research 33 (3): 271–281.
APA
Monfardini, Erica, Paape, M., WANG, Y., Capuco, A., HUSHEEM, M., WOOD, L., & Burvenich, C. (2002). Evaluation of L-selectin expression and assessment of protein tyrosine phosphorylation in bovine polymorphonuclear neutrophil leukocytes around parturition. VETERINARY RESEARCH, 33(3), 271–281.
Vancouver
1.
Monfardini E, Paape M, WANG Y, Capuco A, HUSHEEM M, WOOD L, et al. Evaluation of L-selectin expression and assessment of protein tyrosine phosphorylation in bovine polymorphonuclear neutrophil leukocytes around parturition. VETERINARY RESEARCH. 2002;33(3):271–81.
MLA
Monfardini, Erica, Max Paape, Yan WANG, et al. “Evaluation of L-selectin Expression and Assessment of Protein Tyrosine Phosphorylation in Bovine Polymorphonuclear Neutrophil Leukocytes Around Parturition.” VETERINARY RESEARCH 33.3 (2002): 271–281. Print.