title: Human neutrophil cathepsin G is a potent platelet activator.
abstract: OBJECTIVE
  Neutrophil activation has been implicated in the pathophysiologic condition of ischemia-reperfusion injury, the formation of arterial aneurysms, the progression of myocardial ischemia, and the initiation of deep venous thrombosis. Activated neutrophils release cathepsin G, a serine protease, from their granules, which may cause platelet activation that leads to intravascular thrombosis, tissue infarction, and systemic release of the thrombogenic products of platelet granules. This study used flow cytometry to quantify the extent of cathepsin G-induced platelet activation and degranulation through changes in the expression of platelet surface glycoproteins.
  
  
  METHODS
  Increasing concentrations of human neutrophil-derived cathepsin G were incubated with washed platelets or whole blood from healthy human donors. The platelet surface expression of glycoproteins, including P-selectin, a platelet membrane glycoprotein only expressed after platelet alpha granule release, were determined by quantifying the platelet binding of a panel of fluorescently labeled monoclonal antibodies. Results were compared with the effect of a maximal dose of thrombin, the most potent known platelet activator.
  
  
  RESULTS
  In a washed platelet system, cathepsin G increased platelet surface expression of P-selectin (an activation-dependent neutrophil binding site), the glycoprotein IIb/IIIa complex (fibrinogen receptor), and glycoprotein IV (thrombospondin receptor), and decreased surface expression of glycoprotein Ib (von Willebrand factor receptor) to an extent comparable to maximal thrombin. However, these effects were not observed in a whole blood system. Further experiments revealed that preexposure to plasma completely inhibited cathepsin G-induced washed platelet activation and degranulation. Prostacyclin treatment of washed platelets markedly inhibited cathepsin G-induced platelet activation.
  
  
  CONCLUSIONS
  Cathepsin G is a very potent platelet agonist and degranulator, comparable to maximal thrombin, which alters platelet surface glycoprotein expression for enhanced neutrophil binding and effective platelet aggregation. This study helps to elucidate a possible pathway through which neutrophils may directly activate platelets, leading to intravascular thrombosis, irreversible ischemia, and tissue death in cardiovascular disease states. Patients with diseased endothelium that is deficient in prostacyclin production may be particularly prone to the detrimental effects of neutrophil-derived cathepsin G platelet activation.
authors.affiliation: Department of Surgery, University of Massachusetts Medical School, Worcester 01655.
authors.forename: C A
authors.initials: CA
authors.lastname: LaRosa
authors.forename: M J
authors.initials: MJ
authors.lastname: Rohrer
authors.forename: S E
authors.initials: SE
authors.lastname: Benoit
authors.forename: L J
authors.initials: LJ
authors.lastname: Rodino
authors.forename: M R
authors.initials: MR
authors.lastname: Barnard
authors.forename: A D
authors.initials: AD
authors.lastname: Michelson
mesh_terms: D015703:Antigens, CD / Q000187:drug effects; D018955:CD36 Antigens; D002318:Cardiovascular Diseases / Q000097:blood; D056649:Cathepsin G; D002403:Cathepsins / Q000096:biosynthesis / Q000502:physiology*; D015815:Cell Adhesion Molecules / Q000187:drug effects; D011464:Epoprostenol / Q000494:pharmacology; D005434:Flow Cytometry; D006801:Humans; D009504:Neutrophils / Q000378:metabolism*; D019007:P-Selectin; D015539:Platelet Activation / Q000187:drug effects*; D010974:Platelet Aggregation / Q000187:drug effects; D010980:Platelet Membrane Glycoproteins / Q000187:drug effects*; D012697:Serine Endopeptidases; D013917:Thrombin / Q000494:pharmacology
chemical_list: D015703:Antigens, CD; D018955:CD36 Antigens; D015815:Cell Adhesion Molecules; D019007:P-Selectin; D010980:Platelet Membrane Glycoproteins; D011464:Epoprostenol; D002403:Cathepsins; D012697:Serine Endopeptidases; C534094:CTSG protein, human; D056649:Cathepsin G; D013917:Thrombin