Platelet adhesion at high shear rates: the roles of von Willebrand factor/GPIb and the beta 1 integrin alpha 2 beta 1.
We have previously described a monomeric rvWf fragment, Leu504-Lys728 that contains one disulfide bond linking Cys509-Cys695. This fragment, VCL, has previously been shown to inhibit vWf-ristocetin, asialo-vWf, and botrocetin-induced vWf binding and aggregation of platelets. VCL inhibited 50% of vWf binding to heparin, but it did not inhibit vWf binding to type I collagen. At a high shear force (2600-1 sec), VCL inhibited platelet adhesion to the subendothelial surface of human umbilical arteries. The maximum inhibition of platelet adhesion was 83 +/- 4% at a VCL concentration of 7.6 mumol/L. Various monoclonal anti-Very Late Activation antigens (VLA) antibodies were added to the VCL and tested for their ability to enhance the inhibition of platelet adhesion at high shear forces. Of all of the VLA antibodies tested, only the anti-VLA-2 antibody (176D7) inhibited platelet aggregation in the absence of VCL and enhanced the inhibition of platelet adhesion in the presence of VCL. The VLA-2 antibody and VCL together inhibited 96 +/- 4% of platelet adhesion at high shear forces.
Gralnick
H R
HR
Hematology Service, National Institutes of Health, Bethesda, MD 20892, USA.
Kramer
W S
WS
McKeown
L P
LP
Garfinkel
L
L
Pinot
A
A
Williams
S B
SB
Krutzsch
H
H
D000328:Adult; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000494:pharmacology; D003094:Collagen / Q000378:metabolism; D018056:Hemorheology; D006493:Heparin / Q000378:metabolism; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000276:immunology / Q000494:pharmacology; D007074:Immunoglobulin G / Q000276:immunology / Q000494:pharmacology; D019012:Integrin beta1 / Q000502:physiology*; D016023:Integrins / Q000276:immunology / Q000502:physiology*; D009131:Muscle, Smooth, Vascular / Q000378:metabolism; D010446:Peptide Fragments / Q000494:pharmacology*; D010973:Platelet Adhesiveness / Q000187:drug effects; D010980:Platelet Membrane Glycoproteins / Q000502:physiology*; D011485:Protein Binding / Q000187:drug effects; D011956:Receptors, Cell Surface / Q000502:physiology*; D038981:Receptors, Collagen; D011994:Recombinant Proteins / Q000494:pharmacology; D014469:Umbilical Arteries; D014841:von Willebrand Factor / Q000502:physiology
D000911:Antibodies, Monoclonal; D007140:Immunoglobulin Fab Fragments; D007074:Immunoglobulin G; D019012:Integrin beta1; D016023:Integrins; D010446:Peptide Fragments; D010980:Platelet Membrane Glycoproteins; D011956:Receptors, Cell Surface; D038981:Receptors, Collagen; D011994:Recombinant Proteins; D014841:von Willebrand Factor; C028254:von Willebrand factor receptor; D006493:Heparin; D003094:Collagen