The plasma contact activation system can be viewed as initiating surface-mediated defense reactions. Thus, the biological role of this system is to be viewed primarily as participatory in the pathophysiological reactions to injury, including the coagulation, fibrinolytic, kinin-forming and complement systems, as well as specific interactions with neutrophils and platelets. We wish to delineate the role of two of the contact system proteins. HMW kininogen and kallikrein, in the biochemistry of the inflammatory action. We will probe the importance of proteolytic activation of HMW kininogen in its surface binding activity by using a fluorescent inhibitor assay which avoids proteolytic feedback. We will study the action of this cofactor to inhibit platelet calcium- activated cysteine proteases (calpains) by delineating the kinetics of the reaction and the formation of complexes. We will study genetically engineered inhibitors of contact enzymes, including alphalantitrypsin Pittsburgh and arginine substituted aprotonin. We will use monoclonal antibodies to probe functional domains of contact proteins, as well as to develop new immunochemical assays of neoantigens in enzyme-inhibitor complexes, activated enzymes and activation peptides. These new assays will be applied to patients with hypotensive septicemia, adult respiratory distress syndrome, and cardiopulmonary bypass. We will also further define the role of contact system enzyme, kallikrein as an agonist for stimulation of neutrophils. We will focus on the interaction of fibronectin with these enzymes, the binding of these proteins to neutrophils, and the effect of neutrophis elastase of contact system proteins. This combined biochemical, clinical and cell biological approach should further our understanding of the role of the contact system in inflammation.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL024365-11
Application #
3337651
Study Section
Hematology Subcommittee 2 (HEM)
Project Start
1978-12-01
Project End
1990-11-30
Budget Start
1988-12-01
Budget End
1990-11-30
Support Year
11
Fiscal Year
1989
Total Cost
Indirect Cost
Name
Temple University
Department
Type
Schools of Medicine
DUNS #
City
Philadelphia
State
PA
Country
United States
Zip Code
19122
Veloso, D; Colman, R W (1991) Western blot analyses of prekallikrein and its activation products in human plasma. Thromb Haemost 65:382-8
Pixley, R A; Cassello, A; De La Cadena, R A et al. (1991) Effect of heparin on the activation of factor XII and the contact system in plasma. Thromb Haemost 66:540-7
Kaufman, N; Page, J D; Pixley, R A et al. (1991) Alpha 2-macroglobulin-kallikrein complexes detect contact system activation in hereditary angioedema and human sepsis. Blood 77:2660-7
Colman, R W (1990) Platelet receptors. Hematol Oncol Clin North Am 4:27-42
Bradford, H N; Schmaier, A H; Colman, R W (1990) Kinetics of inhibition of platelet calpain II by human kininogens. Biochem J 270:83-90
Schmaier, A H; Bradford, H N; Lundberg, D et al. (1990) Membrane expression of platelet calpain. Blood 75:1273-81
Colman, R W (1990) Interactions between the contact system, neutrophils and fibrinogen. Adv Exp Med Biol 281:105-20
Clarke, B J; Cote, H C; Cool, D E et al. (1989) Mapping of a putative surface-binding site of human coagulation factor XII. J Biol Chem 264:11497-502
Scott, C F; Colman, R W (1989) Factors influencing the acceleration of human factor XIa inactivation by antithrombin III. Blood 73:1873-9
Gustafson, E J; Schmaier, A H; Wachtfogel, Y T et al. (1989) Human neutrophils contain and bind high molecular weight kininogen. J Clin Invest 84:28-35

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