AT III Barcelona: A Familial Quantitative-Qualitative AT III Deficiency

Summary

A quantitative and qualitative deficiency of antithrombin III (AT III) was found in four members of a Spanish family with thrombotic tendency. In all affected members, levels of AT III antigen and activity (heparin cofactor activity) were reduced to 50% of the normal range. When crossed immunoelectrophoresis (CIE) was performed in the presence of heparin, an abnormal slow-moving peak was found. Crossed immunoelectrofocusing (CIEF) from normal and affected individuals showed that normal AT III migrated between pH 4.9–5.3 while the AT III under study was asymetrically distributed between two pH ranges: 4.9–5.3 and 4.6–4.8. Affinity adsorption of affected members’ plasma to heparin-sepharose beads revealed one population of AT III in the supernatant corresponding to the abnormal AT III, devoid of heparin cofactor activity and showing a peak between pH range: 4.6–4.8 in CIEF.

Our data supports the view that a quantitative-qualitative deficiency was present in the heterozygous state in all the affected family members. Both normal and abnormal ATIII were present in plasma of the affected individuals. This abnormal ATIII was characterized by a lack of affinity for heparin. This familial ATIII deficiency was named ATIII Barcelona.

Key words

Antithrombin III deficiency - Variant - Heparin cofactor - Crossed immunoelectrofocusing - Thrombosis

• References

• 1 Egeberg O. Inherited antithrombin deficiency causing thrombophilia. Thromb Diath Haemorrh 1965; 13: 516-530
• 2 Girolami A, Rubetelli M, Marafiotti F, Zamboni A, Vicarioto MA. Crossed immunoelectrophoresis in the study of abnormal antithrombin III: behavior of ATM Trento. Br J Haematol 1983; 55: 559-561
• 3 Barbui T, Finazzi G, Rodeghiero F, Dini E. Immunoelectrophoresis evidence of a thrombin-induced abnormality in a new variant of hereditary dysfunctional antithrombin III (ATIII “Vicenza”). Br J Haematol 1983; 54: 561-565
• 4 Jorgensen M, Petersen LC, Thorsen S. Purification and characterization of hereditary abnormal antithrombin III with impaired thrombin binding. J Lab Clin Med 1984; 104: 245-256
• 5 Sorensen PJ, Sas G, Peto I, Blasko Gy, Kremmer T, Samu A. Distinction of two pathologic antithrombin III molecules: antithrombin III “Aalborg” and antithrombin III “Budapest”. Thromb Res 1982; 26: 211-219
• 6 Sakuragawa N, Takahashi K, Kondo SI, Koide T. Antithrombin III Toyama: a hereditary abnormal antithrombin III of a patient with recurrent thrombophlebitis. Thromb Res 1983; 31: 305-317
• 7 Chasse JF, Esnard F, Guitton JD, Mouray H, Perigois F, Fauconneau G, Gauthier F. An abnormal plasma antithrombin with no apparent affinity for heparin. Thromb Res 1984; 34: 297-302
• 8 Wolf M, Boyer C, Lavergne JM, Larrieu MJ. A new familial variant of antithrombin III “antithrombin III Paris”. Br J Haematol 1982; 51: 285-295
• 9 Girolami A, Fabris F, Cappellato G, Sainati L, Boeri G. Antithrombin III (ATM) Padua 2: a new congenital abnormality with defective heparin cofactor activities but no thrombotic disease. Blut 1983; 46: 1-4
• 10 Tran TH, Bounameaux H, Bondeli C, Honkanen H, Market GA, Duckert R. Purification and partial characterization of a hereditary abnormal antithrombin III fraction of a patient with recurrent thrombophlebitis. Thromb Haemostas 1980; 44: 87-91
• 11 Sas G, Blasko G, Banhegyi D, Jako J, Palos LA. Abnormal antithrombin III (antithrombin III “Budapest”) as a cause of a familial thrombophilia. Thromb Diath Haemorrh 1974; 32: 105-115
• 12 Bauer KA, Ashenhurst JB, Chédiak J, Rosenberg RD. Antithrombin “Chicago”. A functionally abnormal molecule with increased heparin affinity causing familial thrombophilia. Blood 1983; 62: 1242-1250
• 13 Wolf M, Boyer C, Tripodi A, Meyer D, Larrieu MJ, Mannucci PM. Antithrombin Milano: a new variant with monomeric and dimeric inactive antithrombin III. Blood 1985; 65: 496-500
• 14 Sas G, Peto I, Banhegyi D, Blasko G, Domjan G. Heterogeneity of the “classical” antithrombin III deficiency. Thromb Haemostas 1980; 43: 133-136
• 15 Williams L, Murano G. Human antithrombin III heterogeneity. Blood 1981; 57: 229-232
• 16 Leone G, Cotumaccio R, De Stefano V, Zanetti L. Antithrombin III Roma: a familial quantitative-qualitative AT III deficiency identificable by crossed immunoelectrofocusing and by crossed immunoelec-trophoresis. Haematologica 1983; 68: 765-775
• 17 Caen J, Larrieu MJ, Samama M. L’hemostase. Methodes d’exploration et diagnostic pratique. Paris: l’expansion scientifique; 1975
• 18 Sala N, Owen WG, Collen D. A functional assay of protein C in human plasma. Blood 1984; 63: 671-675
• 19 Laurell CB. Quantitative estimation of protein by electrophoresis in agarose gel containing antibodies. Anal Biochem 1966; 15: 45-52
• 20 Abildgaard U, Lie M, Odegard OR. Antithrombin (heparin cofactor) assay with “new” chromogenic substrates (S-2238 and Chromozym TH). Thromb Res 1977; 11: 549-553
• 21 Odegard OR, Lie M, Abildgaard U. Antifactor Xa activity measured with amidolytic methods. Haemostasis 1976; 5: 265-275
• 22 Sas G, Pepper DS, Cash JD. Investigations on antithrombin III in normal plasma and serum. Br J Haematol 1975; 30: 265-272
• 23 Soderholm J, Smyth CJ, Wadstrom T. A simple and reproducible method of crossed immunoelectrofocusing. Scand J Immunol 1975; 4 (Suppl. 02) 107
• 24 Leone G, Cotumaccio R, De Stefano V, Zanetti L, Bizzi B. Different forms of ATM congenital defect: a study by crossed immunoelectrofocusing. Scand J Haematol 1984; 33: 410-417
• 25 Morse JO. Medical progress: alpha 1-antiptrypsin deficiency. N Engl J Med 1978; 299: 1045-1052
• 26 Fontcuberta J, Grau E, Rubio N, Roig I, Felez J. Quantitative and qualitative congenital deficiency of antithrombin III: a new molecular variant called ATM Barcelona II. Thromb Res (in press)
• 27 Koide T, Odani S, Takahashi K, Ono T, Sakuragawa N. Antihrombin III Toyama: replacement of arginine-47 by cysteine in hereditary abnormal antithrombin III that lacks heparin binding ability. Proc Natl Acad Sci USA 1984; 81: 289-293
• 28 Chang JY, Tran TH. Antithrombin III Basel. Identification of a proleu substitution in hereditary abnormal antithrombin with impaired heparin cofactor activity. J Biol Chem 1986; 261: 1174-1176
• 29 Blackburn MN, Smith RL, Carson J. Sibley. The heparin-binding site of Antithrombin III. Identifiation of a critical tryptophan in the amino acid sequence. J Biol Chem 1984; 259: 939-941
• 30 Villaneuva GB. Predictions of the secondary structure of antithrombin III and the location of the heparin-binding site. J Biol Chem 1984; 259: 2531-2536
• 31 Inomoto T, Shirakami A, Kawauchi S, Shigehiyo T, Saito S, Miyoshi K, Morita T, Iwanaga S. Prothrombin Takushima: characterization of dysfunctional thrombin derived from a variant of human prothrombin. Blood 1987; 69: 565-569
• 32 Prochownik EV, Antonarakis S, Bauer KA, Rosenberg RD, Fearon ER, Orkin SH. Molecular heterogeneity of inherited antithrombin II deficiency. N Engl J Med 1983; 308: 1549-1552
• 33 Duchange N, Chasse JF, Cohen GN, Zakin MM. Molecular characterization of the antithombin III Tours deficiency. Thomb Res 1987; 45: 115-121