Heparan sulfate on activated glomerular endothelial cells and exogenous heparinoids influence the rolling and adhesion of leucocytes.
BACKGROUND
Proliferative glomerulonephritides are characterized by the influx of leucocytes. Heparan sulfate (HS) plays an important role in the recruitment, rolling and firm adhesion of leucocytes to activated endothelium. Recently, we have shown the importance of HS on activated mouse glomerular endothelial cells (mGEnC-1) for the firm adhesion of leucocytes in a static adhesion assay. In the present study, we evaluated the role of HS on glomerular endothelial cells and the effect of adding heparinoids on the leucocyte-glomerular endothelium interaction under dynamic flow conditions.


METHODS
The number of rolling and firmly adhering leucocytes, and the rolling velocity of leucocytes was determined on a monolayer of unactivated or TNF-alpha-activated mGEnC-1 under dynamic flow conditions using physiological relevant shear stress rates in a flow chamber system. Furthermore, the effects of removal of HS on TNF-alpha-activated mGEnC-1 by heparinase III treatment, and of different concentrations of heparin, tinzaparin and HS, on the rolling and adhesion of leucocytes were evaluated.


RESULTS
At the calculated physiological shear stress rate of 0.8 dynes/cm2 the number of rolling and firmly adhering leucocytes to mGEnC-1 increased 2-fold after activation with TNF-alpha, whereas the rolling velocity of the leucocytes decreased 2-fold. Addition of heparin, tinzaparin or HS, and the removal of HS on mGEnC-1 reduced the number of leucocytes rolling and adhering to activated mGEnC-1 about 2-3-fold, while the rolling velocity increased more than 2-fold.


CONCLUSIONS
HS on activated glomerular endothelial cells is important for the interaction with leucocytes under flow conditions, while exogenous heparinoids interfere with this interaction. These results suggest that supplementary treatment of proliferative glomerulonephritides with heparinoids is an interesting option to pursue.
Rops
Angelique L
AL
Nephrology Research Laboratory and Division of Nephrology, Department of Matrix Biochemistry, Nijmegen Centre for Molecular Life Sciences, Radboud University Nijmegen Medical Centre, PO Box 9101, 6500 HB Nijmegen, The Netherlands.
Jacobs
Cor W
CW
Linssen
Peter C
PC
Boezeman
Jan B
JB
Lensen
Joost F
JF
Wijnhoven
Tessa J
TJ
van den Heuvel
Lambert P
LP
van Kuppevelt
Toin H
TH
van der Vlag
Johan
J
Berden
Jo H
JH
D000818:Animals; D000925:Anticoagulants / Q000494:pharmacology*; D002448:Cell Adhesion / Q000187:drug effects; D002450:Cell Communication / Q000187:drug effects; D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects; D004727:Endothelium / Q000187:drug effects / Q000502:physiology*; D005343:Fibrinolytic Agents / Q000494:pharmacology; D006493:Heparin / Q000494:pharmacology; D006495:Heparin, Low-Molecular-Weight / Q000494:pharmacology; D006496:Heparinoids / Q000494:pharmacology*; D006497:Heparan Sulfate / Q000494:pharmacology*; D007678:Kidney Glomerulus / Q000166:cytology* / Q000187:drug effects*; D007962:Leukocytes / Q000166:cytology* / Q000187:drug effects; D051379:Mice; D000078222:Tinzaparin; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
D000925:Anticoagulants; D005343:Fibrinolytic Agents; D006493:Heparin; D006495:Heparin, Low-Molecular-Weight; D006496:Heparinoids; D006497:Heparan Sulfate; D000078222:Tinzaparin; D014409:Tumor Necrosis Factor-alpha