Immunological Studies on the Blood Coagulation Factor X and Its Warfarin-induced Precursor

Summary

Monospecific antisera against the human coagulation factor X have been raised in rabbits by injections of purified antigen. Such antiserum was used to study the cross-reacting material without factor X activity which is present in the blood of warfarin-treated patients and animals as well as to study the changes in factor X during coagulation. One patient with congenital factor X deficiency was also studied.

A complete identity was found between factor X in Macaca mulatta and human blood. During warfarin treatment antigenically cross-reacting material appeared in plasma. This was not adsorbed on BaSO₄, and inhibited the coagulation activity of normal factor X.

Both this material, normal factor X and the cross-reacting material in plasma from a patient congenitally deficient in factor X gave rise to split products during coagulation by the intrinsic pathway, i. e. all of them served as substrates for the intrinsic activator of factor X.

• References

• 1 Bell R. G, Matschiner J. T. Synthesis and Destruction of Prothrombin in the Rat. Archives of Biochemistry and Biophysics 1969; 135: 152
• 2 Denson K. W. E. The Levels of Factors II VII. IX and X by Antibody Neu-tralization Techniques in the Plasma of Patients Receiving Phenindione Therapy. British Journal of Haematology 1971; 20: 643
• 3 Fujikawa K, Legaz M. E, Davie E. W. Bovine Factor X₁ (Stuart Factor). Mechanism of Activation by a Protein from Russell’s Viper Venom. Biochemistry 1972; 11: 4892
• 4 Ganrot P. O, Niléhn J. E. Plasma prothrombin during treatment with dicumarol II. Scandinavian Journal of Clinical and Laboratory Investigation 1968; 22: 23
• 5 Ganrot P. O, Xiléhn J. E. Prothrombin Fragmentation During Coagulation of Whole Blood and Plasma. Scandinavian Journal of Clinical and Laboratory Investigation 1969; 24: 15
• 6 Ganrot P. O, Niléhn J. E. Synthesis of an Abnormal Prothrombin in Malnutrition and Biliary Obstruction and during Dicumarol Treatment. Scandinavian Journal of Clinical and Laboratory Investigation 1971; 28: 245
• 7 Gladhaug Å, Prydz H. Purification of the Coagulation Factors VII and X from human serum. Some properties of factor VII. Biochimica et Biophysica Acta 1970; 215: 105
• 8 Gladhaug Berre Å, Østerud B, Christensen T. B, Holm T, Prydz H. Some Characteristics of the Coagulation Factor Xa purified from Human Serum. Biochemical Journal 1973; 135: 791
• 9 Goodnight S. H, Feinstein D, Østerud B, Rapaport S. I. Factor VII Antibody-neutralizing Material in Hereditary and Acquired Factor VII Deficiency. Blood 1971; 38: 2
• 10 Grossbach U. Acrylamide gel electrophoresis in capillary columns. Biochimica et Biophysica Acta 1965; 107: 180
• 11 Hill R. B, Gaetani S, Paolucci A. M, Rama Rao P. B, Alden R, Ranhotra G. S. Vitamin K and Biosynthesis of Protein and Prothrombin. The Journal of Biological Chemistry 1968; 243: 3930
• 12 Hjort P, Rapaport S. I, Owren P. A. A simple, specific one-stage prothrombin assay using Russell’s viper venom in cephalin suspension. Journal of Laboratory and Clinical Medicine 1955; 46: 89
• 13 Hjort P. F. Intermediate reactions in the coagulation of blood with tissue thromboplastin. Scandinavian Journal of Clinical and Laboratory Investigation 1957; 9 Suppl. 27
• 14 Josso F, Lavergne J. M, Gouault M, Prou-Wartelle O, Soulier P. J. Différents états moléculaires du facteur II. Leur étude a l’aide de la staphylocoagulase et d’anticorps anti-facteur II. Thrombosis et Diathesis Haemorrhagica 1968; 20: 88
• 15 Lanohantln G. F, FRiedmann J. A, Hart D. W. The Conversion of Human Prothrombin to Thrombin by Sodium Citrate. The Journal of Biological Chemistry 1965; 240: 3276
• 16 Larrie U. M. J, Meyer D. Abnormal factor IX during anticoagulant treatment. Lancet 1970; 2: 1085
• 17 Lattre U. C. B. Antigen-Antibody Crossed Electrophoresis. Analytical Bio-chemistry 1965; 14: 358
• 18 LaUrell C. B. Quantitative Estimation of Proteins by Electrophoresis in Agarose Gel Containing Antibodies. Analytical Biochemistry 1966; 15: 45
• 19 Lavergne J. M, Josso F. Tentative immunochemical analysis of the pro-thrombin molecule II. Congress of The International Society on Thrombosis and Haemostasis, Oslo, 12-16 July, 1971 1971. Abstract volume 32
• 20 Lowry O. H, Rosebrough N. J, Farr A. L, Randall R. J. Protein measurements with the Folin phenol reagent. Journal of Biological Chemistry 1951; 193: 265
• 21 Mann K. G, Heldebrant C. M, Fass D. N. Multiple Active Forms of Thrombin I. Partial resolution. Journal of Biological Chemistry 1971; a 246: 5994
• 22 Mann K. G, Heldebrant C. M, Fass D. N. Multiple Active Forms of Thrombin II. Mechanism of Production from Prothrombin. Journal of Biological Chemistry 1971; b 246: 6106
• 23 NilÉhn J. E, Ganrot P. O. Plasma Prothrombin during Treatment with Dicumarol I. Immunochemical Determination of its Concentration in Plasma. Scandinavian Journal of Clinical and Laboratory Investigation 1968; 22: 17
• 24 Owren P. A. The coagulation of blood. Acta Medica Scandinavica 1947; 128 Suppl. 194
• 25 Prydz H. Studies on Proconvertin (Factor VII) IV. The adsorption on barium sulphate. Scandinavian Journal of Clinical and Laboratory Investigation 1964; 16: 409
• 26 Prydz H. Studies on Proconvertin (Factor VII) VI. The production in rabbits of an antiserum against factor VII. Scandinavian Journal of Clinical and Laboratory Investigation 1965; 17: 66
• 27 Prydz H, Gladhaug Å. Factor X Immunological Studies. Thrombosis et Diathesis Haemorrhagica 1971; 25: 157
• 28 Prydz H, Gaudernack G. Studies on the Biosynthesis of Factor VII (proconvertin). The Mode of Action of Warfarin. Biochimica et Biophysica Acta 1971; 230: 373
• 29 Radcliffe R. D, Barton P. G. The Purification and Properties of Activated Factor X. The Journal of Biological Chemistry 1972; 247: 7735
• 30 Rez G, Prydz H. The mode of action of warfarin. An intracellular precursor for Factor VII in rat liver. Biochimica et Biophysica Acta 1971; 244: 495
• 31 Scheidegger J. J. Une micro-methode de l’immuno-electrophorese. International Archives of Allergy 1955; 7: 103
• 32 Shah D. V, SuttIe J. W. Mechanism of Action of Vitamin K: Evidence for the Conversion of a Precursor Protein to Prothrombin in the Rat. Proceedings of the National Academy of Sciences 1971; 68: 1653
• 33 Shah D. V, SUttie J. W. The Effect of Vitamin K and Warfarin on Rat Liver Prothrombin Concentrations. Archives of Biochemistry and Biophysics 1972; 150: 91
• 34 SuttIe J. W. Control of Prothrombin and Factor VII Biosynthesis by Vitamin K. Archives of Biochemistry and Biophysics 1967; 118: 166
• 35 Østerud B, Rapaport S. I. Synthesis of Intrinsic Factor X Activator. Inhibition of the Function of Formed Activator by Antibodies to Factor VIII and to Factor IX. Biochemistry 1970; 9: 1854