Heparan sulfate domains on cultured activated glomerular endothelial cells mediate leukocyte trafficking.
Heparan sulfate (HS) proteoglycans by playing key roles in the leukocyte-endothelial interactions are thought to mediate inflammatory cell influx in proliferative glomerulonephritis. Here, we evaluated the specific features within glomerular endothelial HS that promote leukocyte adhesion. Mouse and human glomerular endothelial cells activated by tumor necrosis factor (TNF)-alpha or interleukin (IL)-1beta increased expression of inflammatory N- and 6-O-sulfated HS domains. In addition, altered expression of HS-modifying enzymes occurred, a feature also found in mouse kidneys with anti-glomerular basement membrane disease or lupus nephritis. Inhibition of the nuclear factor (NF)-kappaB pathway repressed cytokine-induced alterations in HS and gene expression of modifying enzymes. Firm adhesion of leukocytes to activated mouse glomerular endothelial cells decreased after removal of endothelial HS or addition of sulfated heparinoids. Specific antibodies that block N- and 6-O-sulfated HS domains on activated mouse endothelial cells reduced the number of rolling and firmly adhering leukocytes under dynamic flow conditions, while they increased the average leukocyte-rolling velocity. Our study shows that N- and 6-O-sulfated domains in HS on activated glomerular endothelium are crucial for leukocyte trafficking and are possible therapeutic targets.
Rops
A L
AL
Nephrology Research Laboratory, Nijmegen Centre for Molecular Life Sciences, Division of Nephrology, Radboud University Nijmegen Medical Centre, Geert Grooteplein 26, 6525 GA Nijmegen, The Netherlands.
van den Hoven
M J
MJ
Baselmans
M M
MM
Lensen
J F
JF
Wijnhoven
T J
TJ
van den Heuvel
L P
LP
van Kuppevelt
T H
TH
Berden
J H
JH
van der Vlag
J
J
D000818:Animals; D000906:Antibodies / Q000494:pharmacology; D002448:Cell Adhesion / Q000187:drug effects; D002460:Cell Line; D042783:Endothelial Cells / Q000737:chemistry / Q000187:drug effects / Q000276:immunology; D005966:Glucuronidase / Q000378:metabolism; D019812:Heparan Sulfate Proteoglycans / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D053583:Interleukin-1beta / Q000494:pharmacology; D007678:Kidney Glomerulus / Q000737:chemistry / Q000187:drug effects / Q000276:immunology*; D036904:Leukocyte Rolling / Q000187:drug effects; D007962:Leukocytes / Q000276:immunology*; D008181:Lupus Nephritis / Q000276:immunology*; D051379:Mice; D008810:Mice, Inbred C57BL; D016328:NF-kappa B / Q000378:metabolism; D017434:Protein Structure, Tertiary; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D000099175:Heparanase
D000906:Antibodies; D005966:Glucuronidase; D019812:Heparan Sulfate Proteoglycans; D053583:Interleukin-1beta; D016328:NF-kappa B; D014409:Tumor Necrosis Factor-alpha; D000099175:Heparanase