α₁-Antitrypsin Pittsburgh (Met³⁵⁸ → Arg) Inhibits the Contact Pathway of Intrinsic Coagulation and Alters the Release of Human Neutrophil Elastase during Simulated Extracorporeal Circulation

Summary

Cardiopulmonary bypass prolongs bleeding time, increases postoperative blood loss, and triggers activation of plasma proteolytic enzyme systems and blood cells referred to as the “whole body inflammatory response”. Contact of blood with synthetic surfaces leads to qualitative and quantitative alterations in platelets, neutrophils, contact and complement systems. Contact and complement pathway proteins both induce neutrophil activation, a j-antitrypsin Pittsburgh (Met³⁵⁸ → Arg), a mutant of α₁-antitrypsin, is a potent inhibitor of plasma kalli-krein and thrombin. We investigated whether this recombinant mutant protein inhibited platelet activation, as well as contact and/or complement-induced neutrophil activation during simulated extracorporeal circulation.

Arg³⁵⁸ α₁-antitrypsin did not prevent the 34% drop in platelet count at 5 min of recirculation, did not block the 50% decrease in ADP-induced platelet aggregation at 120 min of recirculation, nor inhibit the release of 6.06 α 1.07 µg/ml (3-thromboglobulin at 120 min of recirculation suggesting that the inhibitor had little effect on platelet activation. However, Arg³⁵⁸ α₁ arantitrypsin totally blocked kallikrein-Cl-in-hibitor complex formation but not Cl-Cl-inhibitor complex formation. Most importantly, Arg³⁵⁸ α₁-antitrypsin decreased the release of 1.11 α 0.16 µg/ml human neutrophil elastase by 43%. The attenuation of neutrophil activation in the absence of an effect on complement activation via the classical pathway, supports the concept that kallikrein is a major mediator of neutrophil degranulation during cardiopulmonary bypass.

• References

• Harker LA, Malpass TW, Branson HE, Hessel II EA, Slichter SJ. Mechanism of abnormal bleeding in patients undergoing cardiopulmonary bypass: acquired transient platelet dysfunction associated with selective a-granule release. Blood 1980; 56: 824-834
• 2 Neville WE, Kontaxis A, Gavin T, Clowes Jr GHA. Postperfusion pulmonary vasculitis: its relationship to blood trauma. Arch Surg 1963; 86: 126-137
• 3 Hennessy Jr VL, Hicks RE, Niewiarowski S, Edmunds Jr LH, Colman RW. Function of human platelets during extracorporeal circulation. Am J Physiol 1977; 232: H622-H628
• 4 Dutton RC, Edmunds Jr LH, Hutchinson JC, Roe BB. Platelet aggregate emboli produced in patients during cardiopulmonary bypass with membrane and bubble oxygenators and blood filters. J Thorac Cardiovasc Surg 1974; 67: 258-265
• 5 Wachtfogel YT, Musial J, Jenkin B, Niewiarowski S, Edmunds Jr LH, Colman RW. Loss of platelet β₂-adrenergic receptors during simulated extra-corporeal circulation: prevention with prostaglandin E₁ . J Lab Clin Med 1985; 105: 601-607
• 6 Musial J, Niewiarowski S, Hershock D, Morinelli TA, Colman RW, Edmunds Jr LH. Loss of fibrinogen receptors from the platelet surface during simulated extracorporeal circulation. J Lab Clin Med 1985; 105: 514-522
• 7 Davies GC, Sobel M, Salzman EW. Elevated plasma fibrinopeptide A and thromboxane B₂ levels during cardiopulmonary bypass. Circulation 1980; 61: 808-814
• 8 Beurling-Harbury C, Galvan CA. Acquired decrease in platelet secretory ADP associated with increased postoperative bleeding in post-cardiopulmonary bypass patients and in patients with severe valvular heart disease. Blood 1978; 52: 13-23
• 9 Gnanadurai TV, Branthwaite MA, Colbeck JF, Welman E. Lysosomal enzyme release during cardiopulmonary bypass. Anaesthesia 1977; 32: 743-748
• 10 Wachtfogel YT, Kucich U, Greenplate J, Gluszko P, Abrams W, Weinbaum G, Wenger RK, Rucinski B, Niewiarowski S, Edmunds Jr LH, Colman RW. Human neutrophil degranulation during extracorporeal circulation. Blood 1987; 69: 324-330
• 11 Bolanowski PJP, Bauer J, Machiedo G, Neville WE. Prostaglandin influence on pulmonary intravascular leukocytic aggregation during cardiopulmonary bypass. J Thorac Cardiovasc Surg 1977; 73: 221-224
• 12 Schapira M, Despland E, Scott CF, Boxer LA, Colman RW. Purified human plasma kallikrein aggregates human blood neutrophils. J Clin Invest 1982; 69: 1199-1202
• 13 Wachtfogel YT, Kucich U, James HL, Scott CF, Schapira M, Zimmerman M, Cohen AB, Colman RW. Human plasma kallikrein releases neutrophil elastase during blood coagulation. J Clin Invest 1983; 72: 1672-1677
• 14 Wachtfogel YT, Pixley RA, Kucich U, Abrams W, Weinbaum G, Schapira M, Colman RW. Purified plasma factor XIIa aggregates human neutrophils and causes degranulation. Blood 1986; 67: 1731-1737
• 15 Goldstein IM, Weissmann G. Generation of C5-derived lysosomal enzyme-releasing activity (C5a) by lysates of leukocyte lysosomes. J Immunol 1974; 113: 1583-1588
• 16 Craddock PR, Hammerschmidt D, White JG, Dalmasso AP, Jacob HS. Complement (C5a)-induced granulocyte aggregation in vitro. J Clin Invest 1977; 60: 260-264
• 17 Wachtfogel YT, Harpel PC, Edmunds Jr LH, Colman RW. Formation of CĪ_S-CĪ-inhibitor, kallikrein-C T-inhibitor, and plasmin-α₂-plasmin-inhibitor complexes during cardiopulmonary bypass. Blood 1989; 73: 468-471
• 18 Wachtfogel YT, Kucich U, Hack CE, Gluszko P, Niewiarowski S, Colman RW, Edmunds Jr LH. Aprotinin inhibits the contact, neutrophil, and platelet activation systems during simulated extracorporeal perfusion. J Thorac Cardiovasc Surg 1993; 106: 1-10
• 19 Carrell R, Owen M, Brennan S, Vaughan L. Carboxy ierminal fragment of human α₁-antitrypsin from hydroxy lamine cleavage: homology with antithrombin III. Biochem Biophys Res Commun 1979; 91: 1032-1037
• 20 Courtney M, Jallat S, Tessier L-H, Benavente A, Crystal RG, Lecocq J-P. Synthesis in E.coli of α₁-antitrypsin variants of therapeutic potential for emphysema and thrombosis. Nature 1985; 313: 149-151
• 21 Scott CF, Carrell RW, Glaser CB, Kueppers F, Lewis JH, Colman RW. Alpha-1 -antitrypsin-Pittsburgh: a potent inhibitor of human plasma factor XIa, kallikrein, and factor XII_f . J Clin Invest 1986; 77: 631-634
• 22 Schapira M, Ramus M-A, Jallat S, Carvallo D, Courtney M. Recombinant α₁-antitrypsin Pittsburgh (Met³⁵⁸ → Arg) is a potent inhibitor of plasma kallikrein and activated factor XII fragment. J Clin Invest 1985; 76: 635-637
• 23 Travis J, Matheson NR, George PM, Carrell RW. Kinetic studies on the interaction of α₁-proteinase inhibitor (Pittsburgh) with trypsin-like serine pro-teinases. Biol Chem Hoppe-Seyler 1986; 367: 853-859
• 24 Lewis JH, Iammarino RM, Spero JA, Hasiba U. Antithrombin Pittsburgh: an α₁-antitrypsin variant causing hemorrhagic disease. Blood 1978; 51: 129-137
• 25 Owen MC, Brennan SO, Lewis JH, Carrell RW. Mutation of antitrypsin to antithrombin: a,-antitrypsin Pittsburgh (358 Met → Arg), a fatal bleeding disorder. N Engl J Med 1983; 309: 694-698
• 26 Patston PA, Roodi N, Schifferli JA, Bischoff R, Courtney M, Schapira M. Reactivity of α₁-antitrypsin mutants against proteolytic enzymes of the kallikrein-kinin, complement, and fibrinolytic systems. J Biol Chem 1990; 265: 10786-10791
• 27 Colman RW, Flores DN, DeLa Cadena RA, Scott CF, Cousens L, Barr PJ, Hoffman IB, Kueppers CF, Fisher D, Idell S, Pisarello J. Recombinant α₁-antitrypsin Pittsburgh attenuates experimental gram-negative septicemia. Am J Pathol 1988; 130: 418-426
• 28 Bischoff R, Speck D, Lepage P, Delatre L, Ledoux C, Brown SW, Roitsch C. Purification and biochemical characterization of recombinant α₁-antitrypsin variants expressed in Escherichia coll. Biochemistry 1991; 30: 3464-3472
• 29 Brecher G, Cronkite EP. Morphology and enumeration of human blood platelets. J Appl Physiol 1950; 3: 365-377
• 30 Wintrobe MH. Principles and techniques of blood examination. In: Clinical Hematology Philadelphia: Lea & Febiger; 1967: 410
• 31 Born GVR. Aggregation of blood platelets by adenosine diphosphate and its reversal. Nature 1962; 194: 927-929
• 32 Rucinski B, Niewiarowski S, James P, Walz DA, Budzynski AZ. Antiheparin proteins secreted by human platelets. Purification, characterization, and radioimmunoassay. Blood 1979; 53: 47-62
• 33 Hack CE, Hannema AJ, Eerenberg-Belmer AJM, Out TA, Aalberse RC. A CĪ-inhibitor-complex assay (INCA): a method to detect Cl activation in vitro and in vivo. J Immunol 1981; 127: 1450-1453
• 34 Nuijens JH, Huijbregts CCM, Eerenberg-Belmer AJM, Abbink JJ, Strack van Schijndel RJM, Felt-Bersma RJF, Thijs LG, Hack CE. Quantification of plasma factor Xlla-CĪ-inhibitor and kallikrein-CĪ-inhibitor complexes in sepsis. Blood 1988; 72: 1841-1848
• 35 Rosenberg RD, Damus PS. The purification and mechanism of action of human antithrombin-heparin cofactor. J Biol Chem 1973; 248: 6490-6505
• 36 Kettner C, Mersinger L, Knabb R. The selective inhibition of thrombin by peptides of boroarginine. J Biol Chem 1990; 265: 18289-18297
• 37 Wachtfogel YT, Kettner C, Reilly TM, Knabb RM, Kucich U, Niewiarowski S, Edmunds LH, Colman RW. DuP714, a novel thrombin inhibitor, blocks platelet and neutrophil activation during simulated extracorporeal circulation. Clin Res 1992; 40: 284a
• 38 Gluszko P, Rucinski B, Musial J, Wenger RK, Schmaier AH, Colman RW, Edmunds Jr LH, Niewiarowski S. Fibrinogen receptors in platelet adhesion to surfaces of extracorporeal circuit. Am J Physiol 1987; 252: H615-H621
• 39 Haslam PL, Townsend PJ, Branthwaite MA. Complement activation during cardiopulmonary bypass. Anaesthesia 1980; 35: 22-26
• 40 Chenoweth DE, Cooper SW, Hugli TE, Stewart RW, Blackstone EH, Kirklin JW. Complement activation during cardiopulmonary bypass: evidence for generation of C3a and C5a anaphylatoxins. N Engl J Med 1981; 304: 497-503
• 41 Tauber AI, Kamad AB, Hartshorn KL, Myers JB, Schwartz JH. Parameters of neutrophil activation: models of priming and deactivation. Prog Clin Biol Res 1989; 297: 297-309
• 42 Di Scipio RG. The activation of the alternative pathway C3 convertase by human plasma kallikrein. Immunol 1982; 45: 587-595
• 43 Downing SW, Edmunds LH. Release of vasoactive substances during cardiopulmonary bypass. Ann Thorac Surg 1992; 54: 1236-1243