Platelet adhesion to laminin: role of Ca2+ and Mg2+ ions, shear rate, and platelet membrane glycoproteins.
The adhesion of platelets to purified laminin under flow conditions was investigated. Adhesion to laminin was strongly dependent on the presence of divalent cations. In the absence of cations platelet adhesion (8% coverage in 5 minutes) was maximal at a shear rate of 100/s and no adhesion could be detected at shear rates above 800/s. In the presence of 0.8 mmol/L Mg2+ and 2 mmol/L Ca2+ platelet adhesion reached its maximum (30% coverage) around 800/s. At 1,800/s platelets still adhered to purified laminin (coverage of 6%). Antibodies against the E8 domain of laminin and antibodies against the alpha 6 and beta 1 chains of platelet membrane glycoprotein very late activation antigen-6 (VLA-6), completely inhibited adhesion. No inhibition was found with antibodies against glycoprotein IIb:IIIa, against the alpha 2 chain of VLA-2, and against the alpha 5 chain of VLA-5. Fibronectin and von Willebrand factor were not involved in laminin-dependent adhesion. Anti-VLA-6 partly inhibited platelet adhesion to the extracellular matrix of endothelial cells at shear rates below 800/s. Preincubation of the matrices with antilaminin E8 antibodies did not influence the adhesion. These results show that purified laminin supports platelet adhesion and that the presence of VLA-6 is important for platelet adhesion under flow conditions. The protein in the matrix with which VLA-6 interacts is currently unknown.
Hindriks
G
G
Department of Haematology, University Hospital Utrecht, The Netherlands.
Ijsseldijk
M J
MJ
Sonnenberg
A
A
Sixma
J J
JJ
de Groot
P G
PG
D000327:Adsorption; D000818:Animals; D000906:Antibodies; D001798:Blood Proteins / Q000502:physiology; D002118:Calcium / Q000494:pharmacology*; D002413:Cations, Divalent; D005109:Extracellular Matrix / Q000502:physiology; D005898:Glass; D007797:Laminin / Q000037:antagonists & inhibitors / Q000502:physiology*; D008274:Magnesium / Q000494:pharmacology*; D051379:Mice; D008855:Microscopy, Electron, Scanning; D010973:Platelet Adhesiveness / Q000187:drug effects / Q000502:physiology*; D010980:Platelet Membrane Glycoproteins / Q000037:antagonists & inhibitors / Q000502:physiology*; D016029:Receptors, Very Late Antigen / Q000037:antagonists & inhibitors / Q000502:physiology
D000906:Antibodies; D001798:Blood Proteins; D002413:Cations, Divalent; D007797:Laminin; D010980:Platelet Membrane Glycoproteins; D016029:Receptors, Very Late Antigen; D008274:Magnesium; D002118:Calcium