Hematology-Immunology Module 07 August 2009 Nina Rosario L. Rojas, PhD COAGULATION CASCADE OUTLINE I. II. III. IV. A. B. V. VI. VII. VIII. IX. Blood Components Clotting Factors Steps in Hemostasis Hemostatic Plug Formation Primary: Plate Aggregation Secondary: Coagulation Coagulation Cascade Important Cofactors Clot Removal Tests for Activity Hemophilia C. D. E. I. BLOOD COMPONENTS Fibrinogen stimulates platelet clumping by binding to exposed collagen Activation Platelets release ADP and Thromboxane A2 (an eicosanoid) to activate more platelets Platelets secrete serotonin, phospholipids, lipoproteins and other cascade proteins Aggregation Secondary hemostasis occurs, where cascade forms fibrin mesh or a clot forms to trap platelet ply in place White thrombus (only platelets) or Red Thrombus (with RBCs) Dissolution Plasmin is the key enzyme which dissolves clot to resume blood flow after repair Blood Clotting – Anti-platelet effect of Aspirin is due to platelet cyclooxygenase-1 inhibition. IV. II. CLOTTING FACTORS Immunoglobulins Major Histocompatibility Complexes and Tcell Receptors Cell Surface Markers o Blood Types o HLA Types A. Hemostasis The cessation of blood loss from a damaged blood vessel Blood clotting begins when an injury to the endothelial layer of a blood vessel reveals the collagen in the endothelium Disorders lead to hemorrhage (excessive bleeding) or thrombosis (unnecessary clotting) III. A. B. Group 10 STEPS IN HEMOSTASIS Vascular Constriction Limits blood flow Adhesion Primary hemostasis by platelet plug formation GONZALES. LENON. MENDOZA. PAREJA. SAMSON. HEMOSTATIC PLUG FORMATION A. Primary: Plate Aggregation Plate Aggregation Clotting Collagen-Platelet Interaction Direct: collagen-specific glycoprotein Ia or IIa receptors Indirect: platelets adhere to collagen via the Von Willebrand factors The vWF-platelet interaction is due to glycoproteins Ib, IX and V on the membrane of platelets GP IIIa and IIb also interact with vWF Platelets then release their granule content and form aggregates Von Will ebra nd Page 1 of 4 BATCH 2014 COAGULATION CASCADE D i s e a s e bleeding - Common hereditary disorder Autosomal dominant or autosomal recessive Platelets release their Granule Contents ADP, serotonin, platelet-activating factor (PAF), vWF, platelet factor IV and Thromboxane A2 instigate a Gprotein linked receptor cascade Ca2+ concentration in the cytosol increases, which activates PKC Phospholipase A2 is activated which modifies GP IIb/IIIa Activated platelets change shape from spherical to stellate Clots are formed when fibrinogen cross-links with GP IIb/IIIa - B. - The blood clotting cascade is regulated by positive feedback, or by inhibitors such as antithrombin III (forms an irreversible complex with thrombin) and heparin (enhances antithrombin activity) Plasma concentration of different factors in the plasma that is required for hemostasis may vary to some degree Secondary: Coagulation VI. Coagulation Thrombin Fibrin V. Group 10 COAGULATION CASCADE The conversion of fibrinogen to fibrin via proteolytic cleavage GONZALES. LENON. MENDOZA. PAREJA. SAMSON. Traditionally classified as extrinsic (tissue factors), intrinsic (zymogens and proteases) and common pathways Factors in the blood are synthesized in the liver Numbers refer to order in which factors were discovered, not in order of reaction Cascade is facilitated by proteolytic activation (serine proteases) Fibrinogen is activated to fibrin by IIa (thrombin) which causes fibrin polymerization Thrombin results from activation of II (prothrombin) by Xa The intrinsic pathway seems to be less important, since patients with severe deficiencies of its factors (i.e. FXII do not have a bleeding disorder) IMPORTANT COFACTORS A. Calcium B. Vitamin K Cofactor to hepatic γ-glutamyl carboxylase, which converts carboxyl groups to glutamic acid residues on factors II, VII, IX and X and proteins S, C and Z Activates the said factors and proteins through the oxidation of Vitamin K Regenerated by Vitamin K Epoxide Reductase (VKORC), which reduces it back to its active form Page 2 of 4 BATCH 2014 COAGULATION CASCADE VII. REMOVING THE CLOT Plasmin, a serine protease, whose inactive precursor is plasminogen, has a high affinity for fibrin clots Removal is catalyzed by tissue-type plasminogen activator (TPA), a 72-kd protein that has a domain structure closely related to that of prothrombin VIII. A. B. IX. - Factor VII recently introduced as NovoSeven to control hemorrhage at site of injury (works with tissue factor) TESTS FOR ACTIVITY Contact Factor (Intrinsic) Pathway “contact factors” of plasma Activated partial thromboplastin time test (aPTT) Tissue Factor (Extrinsic) Pathway Tissue factor (specific cellular lipoprotein) Prothrombin time test (PTT) PTT results are often reported as ration to monitor dosing of oral anticoagulants such as warfarin Hemophilia Hemophilia A Factor VIII deficiency X-linked recessive Transfusions are required; however, sometimes Factor VIII is recognized as foreign by the immune system after the process Due to the risk of blood-borne disease, recombinant Factor VIII is now used instead B. Hemophilia B Factor IX deficiency X-linked recessive Called the Christmas disease after the first reported case Transfusions are also used C. Hemophilia C Factor XI deficiency Autosomal recessive Mild bleeding tendencies, such as nose bleeds Recombinant Factor XI or plasma is used NEW THERAPIES A. Group 10 GONZALES. LENON. MENDOZA. PAREJA. SAMSON. Page 3 of 4 BATCH 2014 COAGULATION CASCADE Group 10 GONZALES. LENON. MENDOZA. PAREJA. SAMSON. Page 4 of 4