Targeting platelets in acute experimental stroke: impact of glycoprotein Ib, VI, and IIb/IIIa blockade on infarct size, functional outcome, and intracranial bleeding.
BACKGROUND
Ischemic stroke is a frequent and serious disease with limited treatment options. Platelets can adhere to hypoxic cerebral endothelial cells by binding of their glycoprotein (GP) Ib receptor to von Willebrand factor. Exposure of subendothelial matrix proteins further facilitates firm attachment of platelets to the vessel wall by binding of collagen to their GPVI receptor. In the present study, we addressed the pathogenic role of GPIb, GPVI, and the aggregation receptor GPIIb/IIIa in experimental stroke in mice.


RESULTS
Complete blockade of GPIb alpha was achieved by intravenous injection of 100 microg Fab fragments of the monoclonal antibody p0p/B to mice undergoing 1 hour of transient middle cerebral artery occlusion. At 24 hours after transient middle cerebral artery occlusion, cerebral infarct volumes were assessed by 2,3,5-triphenyltetrazolium chloride staining. In mice treated with anti-GPIb alpha Fab 1 hour before middle cerebral artery occlusion, ischemic lesions were reduced to approximately 40% compared with controls (28.5+/-12.7 versus 73.9+/-17.4 mm3, respectively; P<0.001). Application of anti-GPIb alpha Fab 1 hour after middle cerebral artery occlusion likewise reduced brain infarct volumes (24.5+/-7.7 mm3; P<0.001) and improved the neurological status. Similarly, depletion of GPVI significantly diminished the infarct volume but to a lesser extent (49.4+/-19.1 mm3; P<0.05). Importantly, the disruption of early steps of platelet activation was not accompanied by an increase in bleeding complications as revealed by serial magnetic resonance imaging. In contrast, blockade of the final common pathway of platelet aggregation with anti-GPIIb/IIIa F(ab)2 fragments had no positive effect on stroke size and functional outcome but increased the incidence of intracerebral hemorrhage and mortality after transient middle cerebral artery occlusion in a dose-dependent manner.


CONCLUSIONS
Our data indicate that the selective blockade of key signaling pathways of platelet adhesion and aggregation has a different impact on stroke outcome and bleeding complications. Inhibition of early steps of platelet adhesion to the ischemic endothelium and the subendothelial matrix may offer a novel and safe treatment strategy in acute stroke.
Kleinschnitz
Christoph
C
Department of Neurology, University of Würzburg, Würzburg, Germany.
Pozgajova
Miroslava
M
Pham
Mirko
M
Bendszus
Martin
M
Nieswandt
Bernhard
B
Stoll
Guido
G
D000208:Acute Disease; D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D001792:Blood Platelets / Q000502:physiology; D002543:Cerebral Hemorrhage / Q000401:mortality; D007140:Immunoglobulin Fab Fragments / Q000494:pharmacology; D020244:Infarction, Middle Cerebral Artery / Q000097:blood* / Q000473:pathology / Q000628:therapy*; D002546:Ischemic Attack, Transient / Q000097:blood / Q000473:pathology / Q000628:therapy; D008279:Magnetic Resonance Imaging; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000037:antagonists & inhibitors* / Q000276:immunology; D019038:Platelet Glycoprotein GPIb-IX Complex / Q000037:antagonists & inhibitors* / Q000276:immunology; D010980:Platelet Membrane Glycoproteins / Q000037:antagonists & inhibitors* / Q000276:immunology; D012307:Risk Factors; D015398:Signal Transduction
D000911:Antibodies, Monoclonal; D007140:Immunoglobulin Fab Fragments; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D019038:Platelet Glycoprotein GPIb-IX Complex; D010980:Platelet Membrane Glycoproteins; C098341:platelet membrane glycoprotein VI