Use of heparinase to decrease inflammatory responses

Abstract

Heparinase enzymes can be used as a medical treatment to reduce localized inflammatory responses. Treatment of activated endothelium with heparinase inhibits leukocyte rolling, adhesion and extravasation. Most of the heparin and heparan sulfate on endothelial cell surfaces and in basement membranes is degraded by exposure to heparinase. In addition, immobilized chemokines, which are attached to heparin/heparan sulfate on activated endothelium are solubilized by heparinase digestion. Heparinase can be infused into the vascular system to inhibit accumulation of leukocytes in inflamed tissue and decrease damage resulting from localized inflammations. Targeting of heparinase to activated endothelium can be accomplished through localized administration and/or use of genetically engineered heparinase containing endothelium ligand-binding domains.

1 - 17 . (canceled)  
     
     
         18 . A method to decrease localized inflammatory responses arising from an ischemia/reperfusion injury in a tissue of a human comprising intravascularly administering to said human heparinase enzyme in an effective amount sufficient to decrease neutrophil transmigration through activated endothelium and basement membrane of said vasculature which decreases said localized inflammatory response arising from an ischemia/reperfusion injury.  
     
     
         19 . The method of  claim 18 , wherein said administration of said heparinase enzyme removes and digests heparin and heparan sulfate from endothelial cell surfaces and extracellular matrices of said tissue.  
     
     
         20 . The method of  claim 18 , wherein said administration of said heparinase enzyme decreases the accumulation of leukocytes in tissue adjacent to endothelial cell surfaces and extracellular matrices.  
     
     
         21 . The method of  claim 18 , wherein said administration of said heparinase enzyme inhibits leukocyte extravasation by releasing immobilized chemokines from the endothelium.  
     
     
         22 . The method of  claim 18 , wherein said administration of said heparinase enzyme inhibits leukocyte rolling on endothelium.  
     
     
         23 . The method of  claim 18 , wherein said heparinase enzyme is expressed from a recombinant nucleotide sequence, in  Escherichia coli  or  Flavobacterium heparinum.    
     
     
         24 . The method of  claim 18 , wherein said heparinase enzyme is expressed from a recombinant nucleotide sequence in an organism in which it does not naturally occur.  
     
     
         25 . The method of  claim 18 , wherein said heparinase enzyme is heparinase III.  
     
     
         26 . The method of  claim 18 , wherein said ischemia/reperfusion injury is selected from the group consisting of myocardial infarction, stroke, organ transplant, traumatic shock, cardiovascular surgery.