PMID- 9310484
OWN - NLM
STAT- MEDLINE
DCOM- 19971020
LR  - 20210216
IS  - 0006-4971 (Print)
IS  - 0006-4971 (Linking)
VI  - 90
IP  - 6
DP  - 1997 Sep 15
TI  - Relative importance of the glycoprotein Ib-binding domain and the RGD sequence of 
      von Willebrand factor for its interaction with endothelial cells.
PG  - 2335-44
AB  - Endothelial cell adhesion to von Willebrand Factor is mainly mediated through an 
      interaction between the alpha vbeta3 integrin and the RGD sequence of von 
      Willebrand factor (vWF). To define the potential involvement of glycoprotein Ib 
      alpha (GPIb alpha) as an endothelial vWF receptor, we compared cell adhesion to 
      three recombinant vWF, the wild-type (WT-rvWF) and two mutants, RGGS-rvWF 
      (D1746G), defective for binding to platelet alphaIIb beta3, and deltaA1-rvWF with 
      a deletion between amino-acids 478 and 716, which does not bind to platelet GPIb 
      alpha. Adhesion of human umbilical vein endothelial cells to purified vWF 
      recombinants was measured by automatized cell counting using an image analyzer. 
      Whereas cell adhesion to delta A1-rvWF was unchanged compared with WT-rvWF, 
      reaching a plateau of 40% total cells at a concentration of 2.5 microg/mL rvWF, 
      adhesion to RGGS-rvWF was only 10% of total cells. Cell stimulation by tumor 
      necrosis factor-alpha (TNF alpha), reported to upregulate the expression of the 
      putative endothelial GPIb alpha, did not modify adhesion to these rvWF. 
      Monoclonal antibodies to vWF or GPIb alpha, blocking vWF interaction with 
      platelet GPIb alpha, were unable to inhibit endothelial cell adhesion to rvWF. In 
      contrast, antibody 9 to vWF, blocking the alpha vbeta3-dependent endothelial cell 
      adhesion to plasma vWF, inhibited adhesion to WT-rvWF as efficiently as to 
      deltaA1-rvWF (50% inhibition at a concentration of 11 and 15 microg/mL, 
      respectively). In agreement with the fact that endothelial cell adhesion to vWF 
      appeared independent of the GPIb alpha-binding domain, we were unable to detect 
      endothelial surface expression of GPIb alpha by flow cytometry or in cell lysates 
      by immunoprecipitation followed by immunoblotting. Moreover, expression of GPIb 
      alpha mRNA was undetectable in endothelial cells, even after stimulation by TNF 
      alpha. These studies indicate that GPIb alpha is not expressed in human cultured 
      endothelial cells and is not involved in adhesion to vWF-containing surfaces. 
      Thus, in static conditions, cultured endothelial cells adhere to vWF through an 
      alpha vbeta3-dependent, GPIb alpha-independent mechanism.
FAU - Perrault, C
AU  - Perrault C
AD  - INSERM U143, Hopital de Bicetre, France.
FAU - Lankhof, H
AU  - Lankhof H
FAU - Pidard, D
AU  - Pidard D
FAU - Kerbiriou-Nabias, D
AU  - Kerbiriou-Nabias D
FAU - Sixma, J J
AU  - Sixma JJ
FAU - Meyer, D
AU  - Meyer D
FAU - Baruch, D
AU  - Baruch D
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Blood
JT  - Blood
JID - 7603509
RN  - 0 (Antibodies, Monoclonal)
RN  - 0 (Cell Adhesion Molecules)
RN  - 0 (Oligopeptides)
RN  - 0 (Peptide Fragments)
RN  - 0 (Platelet Glycoprotein GPIb-IX Complex)
RN  - 0 (RNA, Messenger)
RN  - 0 (Tumor Necrosis Factor-alpha)
RN  - 0 (von Willebrand Factor)
RN  - 78VO7F77PN (arginyl-glycyl-aspartic acid)
SB  - IM
MH  - Antibodies, Monoclonal/immunology
MH  - Binding Sites
MH  - Cell Adhesion/drug effects
MH  - Cell Adhesion Molecules/chemistry/metabolism
MH  - Cells, Cultured
MH  - Endothelium, Vascular/*metabolism
MH  - Gene Expression
MH  - Humans
MH  - Oligopeptides
MH  - Peptide Fragments
MH  - Platelet Glycoprotein GPIb-IX Complex/chemistry/*metabolism
MH  - RNA, Messenger/genetics
MH  - Tumor Necrosis Factor-alpha/pharmacology
MH  - von Willebrand Factor/chemistry/*metabolism
EDAT- 1997/10/06 00:00
MHDA- 1997/10/06 00:01
CRDT- 1997/10/06 00:00
PHST- 1997/10/06 00:00 [pubmed]
PHST- 1997/10/06 00:01 [medline]
PHST- 1997/10/06 00:00 [entrez]
AID - S0006-4971(20)58028-2 [pii]
PST - ppublish
SO  - Blood. 1997 Sep 15;90(6):2335-44.