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NDT Advance Access originally published online on January 25, 2007
Nephrology Dialysis Transplantation 2007 22(4):1070-1077; doi:10.1093/ndt/gfl801
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© The Author [2007]. Published by Oxford University Press on behalf of ERA-EDTA. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org

Heparan sulfate on activated glomerular endothelial cells and exogenous heparinoids influence the rolling and adhesion of leucocytes

Angelique L. Rops1, Cor W. Jacobs1, Peter C. Linssen2, Jan B. Boezeman2, Joost F. Lensen3, Tessa J. Wijnhoven3,4, Lambert P. van den Heuvel4, Toin H. van Kuppevelt3, Johan van der Vlag1 and Jo H. Berden1

1Nephrology Research Laboratory and Division of Nephrology, 2Central Hematology Laboratory, 3Department of Matrix Biochemistry and 4Department of Pediatrics, Nijmegen Centre for Molecular Life Sciences, Radboud University Nijmegen Medical Centre, Nijmegen, The Netherlands

Correspondence and offprint requests to: Jo H. M. Berden, MD, PhD, Division of Nephrology (464), Radboud University Nijmegen Medical Centre, PO Box 9101, 6500 HB Nijmegen, The Netherlands. Email: j.berden{at}nier.umcn.nl



  Abstract

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.

Keywords: adhesion of leucocytes; glomerular endothelial cells; glomerulonephritis; heparin; low molecular weight heparin; rolling of leucocytes

Received for publication: 11. 8.06
Accepted in revised form: 8.12.06


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