Dissolution of arterial platelet thrombi in vivo with a bifunctional platelet GPIIIa49-66 ligand which specifically targets the platelet thrombus.
Patients with HIV-1 immune-related thrombocytopenia have a unique antibody (Ab) against integrin GPIIIa49-66 capable of inducing oxidative platelet fragmentation via Ab activation of platelet nicotinamide adenine dinucleotide phosphate oxidase and 12-lipoxygenase releasing reactive oxygen species. Using a phage display single-chain antibody (scFv) library, we developed a novel human monoclonal scFv Ab against GPIIIa49-66 (named A11) capable of inducing fragmentation of activated platelets. In this study, we investigated the in vivo use of A11. We show that A11 does not induce significant thrombocytopenia or inhibit platelet function. A11 can prevent the cessation of carotid artery flow produced by induced artery injury and dissolve the induced thrombus 2 hours after cessation of blood flow. In addition, A11 can prevent, as well as ameliorate, murine middle cerebral artery stroke, without thrombocytopenia or brain hemorrhage. To further optimize the antithrombotic activity of A11, we produced a bifunctional A11-plasminogen first kringle agent (SLK), which homes to newly deposited fibrin strands within and surrounding the platelet thrombus, reducing effects on nonactivated circulating platelets. Indeed, SLK is able to completely reopen occluded carotid vessels 4 hours after cessation of blood flow, whereas A11 had no effect at 4 hours. Thus, a new antithrombotic agent was developed for platelet thrombus clearance.
Department of Medicine, New York University School of Medicine, New York, NY 10016, USA.
Wei
W
Zhang
Yong-Sheng
YS
Li
Michael A
MA
Nardi
Suying
S
Dang
Jing
J
Yang
Yong
Y
Ji
Zongdong
Z
Li
Simon
S
Karpatkin
Thomas
T
Wisniewski
D000818:Animals; D018033:Antibodies, Bispecific / Q000235:genetics / Q000627:therapeutic use*; D001483:Base Sequence; D002341:Carotid Artery Thrombosis / Q000097:blood / Q000628:therapy; D017931:DNA Primers / Q000235:genetics; D005343:Fibrinolytic Agents / Q000627:therapeutic use*; D006801:Humans; D066298:In Vitro Techniques; D020244:Infarction, Middle Cerebral Artery / Q000097:blood / Q000517:prevention & control / Q000628:therapy; D008024:Ligands; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D010446:Peptide Fragments / Q000037:antagonists & inhibitors* / Q000276:immunology / Q000378:metabolism; D019151:Peptide Library; D015539:Platelet Activation / Q000187:drug effects; D010974:Platelet Aggregation / Q000187:drug effects; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000037:antagonists & inhibitors* / Q000276:immunology / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000627:therapeutic use; D013927:Thrombosis / Q000097:blood* / Q000628:therapy*
D018033:Antibodies, Bispecific; D017931:DNA Primers; D005343:Fibrinolytic Agents; D008024:Ligands; D010446:Peptide Fragments; D019151:Peptide Library; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D011994:Recombinant Proteins; C106958:platelet glycoprotein (GP)IIIa 49-66