title: Human platelet glycoprotein VI function is antagonized by monoclonal antibody-derived Fab fragments.
abstract: Platelet interactions with adhesive ligands exposed at sites of vascular injury initiate the normal hemostatic response but may also lead to arterial thrombosis. Platelet membrane glycoprotein (GP)VI is a key receptor for collagen. Impairment of GPVI function in mice results in a long-term antithrombotic protection and prevents neointimal hyperplasia following arterial injury. On the other hand, GPVI deficiency in humans or mice does not result in serious bleeding tendencies. Blocking GPVI function may thus represent a new and safe antithrombotic approach, but no specific, potent anti-GPVI directed at the human receptor is yet available. Our aim was to produce accessible antagonists of human GPVI to evaluate the consequences of GPVI blockade. Amongst several monoclonal antibodies to the extracellular domain of human GPVI, one, 9O12.2, was selected for its capacity to disrupt the interaction of GPVI with collagen in a purified system and to prevent the adhesion of cells expressing recombinant GPVI to collagen and collagen-related peptides (CRP). While 9O12.2 IgGs induced platelet activation by a mechanism involving GPVI and Fc gamma RIIA, 9O12.2 Fab fragments completely blocked collagen-induced platelet aggregation and secretion from 5 microg mL-1 and fully prevented CRP-induced activation from 1.5 microg mL-1. 9O12.2 Fabs also inhibited the procoagulant activity of collagen-stimulated platelets and platelet adhesion to collagen in static conditions. Furthermore, 9O12.2 Fabs impaired platelet adhesion, and prevented thrombi formation under arterial flow conditions. We thus describe here for the first time a functional monoclonal antibody to human GPVI and demonstrate its effect on collagen-induced platelet aggregation and procoagulant activity, and on thrombus growth.
authors.affiliation: INSERM E348, Faculté de Médecine Xavier Bichat, Paris, France.
authors.forename: C
authors.initials: C
authors.lastname: Lecut
authors.forename: L A
authors.initials: LA
authors.lastname: Feeney
authors.forename: G
authors.initials: G
authors.lastname: Kingsbury
authors.forename: J
authors.initials: J
authors.lastname: Hopkins
authors.forename: F
authors.initials: F
authors.lastname: Lanza
authors.forename: C
authors.initials: C
authors.lastname: Gachet
authors.forename: J-L
authors.initials: JL
authors.lastname: Villeval
authors.forename: M
authors.initials: M
authors.lastname: Jandrot-Perrus
mesh_terms: D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D003094:Collagen / Q000378:metabolism; D003435:Crotalid Venoms / Q000378:metabolism; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000494:pharmacology*; D037181:Lectins, C-Type / Q000378:metabolism; D010477:Perfusion; D015539:Platelet Activation / Q000187:drug effects; D010973:Platelet Adhesiveness / Q000187:drug effects; D010980:Platelet Membrane Glycoproteins / Q000037:antagonists & inhibitors* / Q000502:physiology; D013927:Thrombosis / Q000517:prevention & control
chemical_list: D000911:Antibodies, Monoclonal; D003435:Crotalid Venoms; D007140:Immunoglobulin Fab Fragments; D037181:Lectins, C-Type; D010980:Platelet Membrane Glycoproteins; C098341:platelet membrane glycoprotein VI; C028890:convulxin; D003094:Collagen