Some Properties of the Anticoagulant Factor of Aged Pooled Plasma^[*]

Summary

Hematologic studies of an infant who was admitted for shock and was treated with a large volume of pooled aged plasma disclosed striking prolongation of a variety of coagulation tests, including the prothrombin time, partial thromboplastin time, and thrombin time. The aged plasma produced a marked anticoagulant effect on normal plasma in vitro with the results suggesting inhibition of the thrombin-fibrinogen reaction. Similar results were obtained with 24 different lots of aged plasma from 5 different sources. The inhibitory factor of aged plasma behaved as a competitive inhibitor of the conversion of fibrinogen to fibrin, as compared with heparin which is a noncompetitive inhibitor. Infusion of aged plasma into rabbits produced a defect similar to that found in the patient. Biochemical and immunologic studies revealed that the plasma was devoid of fibrinogen but contained large amounts of a fibrinogen degradation product. Treatment of the plasmas with protamine or with rabbit IgG anti-human fibrinogen removed the anticoagulant activity. Development of anticoagulant activity during plasma aging did not appear to be related to fibrinolysis, as it was unaffected by addition of a fibrinolytic inhibitor. Because of the anticoagulant activity of aged plasma, it must be used with caution in patients having underlying hemostatic disorders. Coagulation screening tests in patients who have received recent infusions of aged plasma may lead to misinterpretation unless specific coagulation factor assays are performed.

^* A preliminary report of these studies was presented at the thirtyeight annual meeting of the Society for Pediatric Research, May 4, 1968 at Atlantic City, New Jersey.

^** Address: Department of Pediatrics, 840 South Wood Street, Chicago, Illinois, 60612.

• References

• 1 Taylor F. H. L, Lozner E. L, Davidson G. S, Tagnon H. J, Newhouser L. R. Preservation of normal human plasma in the liquid state. II. Comparative in vitro studies on the physiologic activity of labile constituents of liquid and frozen plasma. J. clin. Invest. 23: 351 1944;
• 2 Ratnoff O. E, Menzie G. New method for determination of fibrinogen in small samples of plasma. J. Lab. clin. Med. 37: 316 1951;
• 3 Quick A. J. Hemorrhagic Diseases. 379 Lea and Febiger; Philadelphia: 1957
• 4 Miale J. B. The fibrometer system for routine coagulation tests. Prothrombin time and partial thromboplastin time, macro and micro. Amer. J. clin. Path. 43: 475 1965;
• 5 Hardisty R. M, Ingram G. I. G. Bleeding Disorders. Investigation and Management. 285 F. A. Davis; Philadelphia: 1965
• 6 Gwren P. A, Aas K. Control of dicumarol therapy and quantitative determination of prothrombin and proconvertin. Scand. J. clin. Lab. Invest. 3: 201 1951;
• 7 Shanberge J. N, Matsuoka T, Fukui H. A simple method for the preparation of a substrate for a one-stage factor V assay. Techn. Bull. Regist. Med. Techn. 37: 67 1967;
• 8 Simone J. V, Vanderheiden J, Abildgaard G. F. A semiautomatic one-stage factor VIII assay with a commercially prepared standard. J. Lab. clin. Med. 69: 706 1967;
• 9 Gross S, Keefer V, Newman A. J. Platelets in iron deficiency and response to therapy. Pediatrics 34: 315 1964;
• 10 Rodriguez-Erdmann F. Bleeding due to increased intravascular blood coagulation: hemorrhagic syndromes caused by consumption of blood-clotting factors (consumption coagulopathies). New Engl. J. Med. 273: 1370 1965;
• 11 Lineweaver H, Burk D. The determination of enzyme dissociation constants. J. Amer. chem. Soc. 56: 658 1934;
• 12 Fletcher A. P, Alkjaersig N, Sherry S. Pathogenesis of the coagulation defect developing during pathological plasma proteolytic (“fibrinolytic”) states. I. The significance of fibrinogen proteolysis and circulating fibrinogen breakdown products. J. clin. Invest. 41: 896 1962;
• 13 Dixon M. The determination of enzyme inhibitor constants. Biochem. J. 55: 170 1953;
• 14 Larrieu M. J, Marder V. J, Inceman S. Effects of fibrinogen degradation products on platelets and coagulation. Thrombos. Diathes. haemorrh. (Stuttg.) Suppl.20: 215 1967;
• 15 Sober H. A, Gutter F. J, Wyckoff M. M, Peterson E. A. Chromatography of proteins. II. Fractionation of serum protein on anion-exchange cellulose. J. Amer. chem. Soc. 78: 756 1956;
• 16 Kopec M, Kowalski E, Stachurska J. Studies on paracoagulation role of antithrombin VI. Thrombos. Diathes. haemorrh. (Stuttg.) 5: 285 1960;
• 17 Lipinski B, Wegrznowicz Z, Budzynski A, Kopec M, Batallo Z. S, Kowalski E. Soluble unclottable complexes formed in the presence of fibrinogen degradation products (FDP) during the fibrinogen-fibrin conversion and their potential significance in pathology. Thrombos. Diathes. haemorrh. (Stuttg.) 17: 65 1967;
• 18 Alkjaersig N, Fletcher A, Sherry S. E-Aminocaproic acid: an inhibitor of plasminogen activation. J. biol. Chem. 234: 832 1959;
• 19 Searcy R. L, Simms N. M, Low E. M. Y. A simple method for rapid detection of hypo-fibrinogenemia. Amer. J. med. Tech. 33: 326 1967;
• 20 Seegers W. H, Nieft M. L, Vandenbelt J. M. Decomposition products of fibrinogen and fibrin. Arch. Biochem. 7: 15 1945;
• 21 Stefanini M. Antithrombin activity of stored plasma. Amer. J. clin. Path. 18: 537 1948;
• 22 Stormorken H. Reactivity of stored plasma to thrombin, with reference to the fibrinogen conversion accelerator and heparinoid activity. Brit. J. Haemat. 3: 299 1957;
• 23 Triantaphyllopoulos D. C. Anticoagulant effect of incubated fibrinogen. Canad. J. Biochem. 56: 249 1958;
• 24 Cutler S. S, Burbank B, Marzullo E. R. Hypocoagulability of certain irradiated plasmas. J. Amer. med. Ass. 115: 1057 1950;
• 25 Tanturi C. A, and Wetzel N. C. Studies upon the relation between plasma antithrombin and heparin. Amer. J. med. Sci. 217: 410 1949;
• 26 Triantaphyllopoulos D. C, Quick A. J, Greenwalt T. J. Action of disodium ethylenediamine tetracetate on blood coagulation. Evidence of the development of heparinoid activity during incubation or aeration of plasma. Blood 10: 534 1955;
• 27 Triantaphyllopoulos D. C, Muirhead C. R. Formation of anticoagulants by digesting fibrin with thrombin. Thrombos. Diathes. haemorrh. (Stuttg.) 19: 397 1968;