Cytokine treatment of endothelial cells increases glycoprotein Ib alpha-dependent adhesion to von Willebrand factor.
Endothelial cells (EC) possess at least two membrane receptors for von Willebrand factor (vWF), the vitronectin receptor (VNR, alpha(v)beta3), which recognizes an Arg-Gly-Asp (RGD) sequence in the C-terminus of vWF, and glycoprotein Ib alpha (GP Ib alpha), which interacts with a region in the N-terminal A1 domain of vWF. In the absence of added cytokines, EC attachment to a vWF substratum is mediated largely through the alpha(v)beta3, with a smaller contribution by GP Ib alpha. In the present study, we have examined the effect of cytokines on the receptor specificity of EC attachment to wild-type vWF (WT-vWF) and to vWF, which had been mutated in the C-terminal RGDS sequence (RADS-vWF). Exposure of human umbilical vein EC (HUVEC) to tumor necrosis factor-alpha (TNF-alpha) or to TNF-alpha in combination with interferon-gamma (IFN-gamma), but not to interleukin-1beta (IL-1), increased attachment to RADS-vWF by about twofold. The TNF-alpha-induced increase in EC attachment was accompanied by an increase in cell surface GP Ib alpha expression; GP Ib alpha surface expression was not increased by IL-1. Attachment of untreated HUVEC to WT-vWF could be inhibited 60% to 70% by a monoclonal antibody (MoAb) (LM609) to the VNR and 30% to 40% by the A1 fragment of vWF (containing the GP Ib alpha binding domain). The pattern of inhibition of attachment to WT-vWF was largely unchanged after TNF-alpha treatment of HUVEC. In contrast, the attachment to WT-vWF of HUVEC, treated with TNF-alpha +IFN-gamma was completely inhibited by vWF-A1 and inhibited only 35% by the anti-VNR antibody LM609. Two MoAbs to GP Ib alpha produced similar, but incomplete, inhibition. Pretreatment of HUVEC with the combination of TNF-alpha + IFN-gamma produced a dramatic decrease in VNR expression, confirming previous findings of Defilippi et al. These results suggest that in the presence of the inflammatory cytokines TNF-alpha + IFN-gamma, the endothelial GP Ib complex is a major determinant of HUVEC adhesion to surface-bound vWF.
Beacham
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DA
The Cardeza Foundation for Hematologic Research, Department of Medicine, Jefferson Medical College of Thomas Jefferson University, Philadelphia, PA 19107-5099, USA.
Tran
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LP
Shapiro
S S
SS
D002448:Cell Adhesion / Q000187:drug effects; D002478:Cells, Cultured; D016207:Cytokines / Q000494:pharmacology*; D004730:Endothelium, Vascular / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D019038:Platelet Glycoprotein GPIb-IX Complex / Q000378:metabolism*; D011994:Recombinant Proteins / Q000494:pharmacology; D014841:von Willebrand Factor
D016207:Cytokines; D019038:Platelet Glycoprotein GPIb-IX Complex; D011994:Recombinant Proteins; D014841:von Willebrand Factor