Use of Factor VIII in the Treatment of Hemophilia

1962 ◽  
Vol 07 (02) ◽  
pp. 230-238 ◽  
Author(s):  
A Pavlovsky ◽  
H Peterson ◽  
G Casillas ◽  
C Simonetti ◽  
A Martinez Canaveri ◽  
...  

SummaryThis publication describes the results obtained by treatment of haemophiliacs with factor VIII preparations isolated from Cohn fraction I by use of tannic acid, FI-O-Ta.The authors stress the rapidity of the disappearance of factor VIII after injection. Transfusions are generally well tolerated. One reaction of the pyrogen type has been observed and also a case of activation of the fibrinolytic system.A second purification method by means of chromatography on DEAE-cellulose is described.

1998 ◽  
Vol 18 (02) ◽  
pp. 74-79
Author(s):  
K.-H. Zurborn ◽  
H. D. Bruhn ◽  
H. Mönig

SummaryIn order to study the acute and prolonged effects of hypoglycemia on the hemostatic and fibrinolytic system we measured prothrombin fragment (F1+2), thrombin-antithrombin III complex (TAT), platelet factor 4 (PF4), β-thromboglobulin (âTG), factor VIII antigen (F VIII antigen), D-dimer, tissue-type plasminogen activator (t-PA) antigen, and plasminogen activator inhibitor (PAI-1) in 22 patients during insulin tolerance test. F1+2 and TAT increased significantly 15 and 90 minutes after administration of insulin, as did PF4 and âTG. At 4 and 24 hours, these parameters were not different from baseline. Factor VIII antigen was not significantly altered. D-dimer concentration did not change. However, the D-dimer/TAT ratio significantly decreased at 15 and 90 minutes but increased markedly above baseline at 4 and 24 hours. t-PA antigen was also found to be elevated at 15 and 90 minutes but had returned to baseline at 4 and 24 hours. PAI-1 concentration did not change. We conclude from these data that both coagulation and fibrinolysis are activated in the short-term response to acute insulin-induced hypoglycemia, followed by a prolonged activation of fibrinolysis. Our study may explain why patients undergoing insulin tolerance test, despite marked clotting and platelet activation, almost never develop thromboembolic complications.


Blood ◽  
1970 ◽  
Vol 35 (6) ◽  
pp. 809-820 ◽  
Author(s):  
LEON W. HOYER ◽  
ROBERT T. BRECKENRIDGE

Abstract Patients with a genetic variant of hemophilia A have in their plasmas material which has antigenic characteristics similar to those of antihemophilic factor (AHF). The physical properties of the biologically inactive cross-reacting material (CRM) are like those of AHF from normal plasma. The CRM is concentrated in Cohn fraction I and in cryoprecipitates and is not adsorbed from plasma by calcium phosphate or barium sulfate. It is inactivated by heating to 56° for 30 minutes. The CRM is less sensitive to thrombin inactivation than AHF and is recovered in serum. The similar properties of AHF and CRM support the hypothesis that patients with this genetic variant of hemophilia A synthesize a material similar to AHF but lacking procoagulant activity.


Blood ◽  
1967 ◽  
Vol 30 (2) ◽  
pp. 137-150 ◽  
Author(s):  
HERBERT S. STRAUSS ◽  
EZIO MERLER

Abstract A circulating anticoagulant, which specifically inhibits Factor VIII (AHF), has been detected in some patients with hemophilia A who had received multiple transfusions. The inhibitor was quantitated by measurement of the degree of inactivation of Factor VIII. The data presented provide strong evidence for the antibody nature of the Factor VIII inhibitor in hemophilia: (1) All specific inhibitory activity of serum was detected in the γG globulin obtained by chromatography of the sera on DEAE cellulose. (2) Fab fragments obtained by digestion of the γG globulin with papain, contained 18-22 per cent of the specific inhibitory activity, while Fc fragments contained 0.4-3 per cent. F(ab')2 fragments obtained by digestion with pepsin contained 36-61 per cent of the specific inhibitory activity of γG globulin. (3) The level of the inhibitor of Factor VIII increased sharply following transfusions of blood and decreased slowly to its preinfusion level. (4) When small amounts of inhibitor were incubated in vitro with excess Factor VIII, the inhibitor activity was decreased. Infusion of Factor VIII into a patient with a low level of inhibitor decreased the inhibitor activity. (5) Clearance of the isologous inhibitor from the circulation of a normal subject was rapid.


1966 ◽  
Vol 15 (03/04) ◽  
pp. 338-342
Author(s):  
S van Creveld ◽  
I. A Mochtar ◽  
C. N Pascha

SummaryThe anti-hemophilic factor (AHF, factor VIII) has been separated from fibrinogen by means of continous flow electrophoresis. The starting material was Cohn’s fraction I, prepared from normal human resinplasma. These fibrinogen-free products still contained proteins without AHF-activity. Part of these contaminating proteins could be extracted from fraction I by means of a 1 molar solution of epsilon amino caproic acid. The residue, principally consisting of fibrinogen and AHF, was dissolved in a triethylamine buffer and subjected to continuous flow electrophoresis. The resulting AHF-preparations were free from fibrinogen. By immuno-electrophoresis no proteins without AHF-activity could be demonstrated in these preparations.


1961 ◽  
Vol 6 (02) ◽  
pp. 287-307 ◽  
Author(s):  
W. F Stapp

SummaryAdequate fibrinogen reactivity is suggested as an additional requirement for specific fibrinogen reagents, to be added to the well known requirements of protein concentration, optimum pH, and optimum ionic strength. Reasons and a method for testing are outlined.A method for the preparation of a specific fibrinogen reagent, deficient in factor VIII, from BaSO4 adsorbed bovine plasma is described in detail. The method described utilizes either fresh or regenerated DEAE cellulose as an adsorbent in a non-column chromatographic adsorption procedure, associated with a dialysis procedure in which the final fibrinogen product is concentrated and precipitated as a “euglobulin”. The “euglobulin” fibrinogen precipitate was then dissolved/suspended in buffered saline at a specific pH and NaCl concentration. Essentially the method can be briefly outlined into 5 separate steps:1. Dialysis of BaSO4 adsorbed bovine plasma against 0.5% NaCl at pH 6.6 in 0.005 M Imidazole-HCl buffer.2. Washing of DEAE cellulose with the same type of buffered NaCl solution to specific conditions.3. Mixing of the dialyzed BaSO4 adsorbed bovine plasma with the washed DEAE cellulose and separation by centrifugation.4. Concentration of the cellulose adsorbed plasma as a “euglobulin” fibrinogen precipitate by dialysis against running tap water.5. Dissolving of the “euglobulin” fibrinogen precipitate in 0.9% NaCl buffered at pH 6.8.Fibrinogen preparations, prepared with the use of regenerated DEAE cellulose were found to be similar to those prepared by fresh DEAE cellulose in fibrinogen reactivity and deficiency in factor VIII activity, but were found to have less fibrinogen concentration than those prepared using fresh DEAE cellulose.Suggestions were also made for possibly increasing the over-all yield of the fibrinogen reagent through use of a “washing-elution” procedure.The procedure that was used for regenerating the DEAE cellulose was also described in detail and commented upon.


1967 ◽  
Vol 17 (01/02) ◽  
pp. 188-193 ◽  
Author(s):  
S van Creveld ◽  
I. A Mochtar ◽  
C. N Pascha

SummaryPreparations of normal human antihemophilic factor (AHF, factor VIII) obtained by continuous free electrophoresis in the “Elphor” apparatus, were investigated for enzymatic activity on various substrates. A number of enzymatic activities could be excluded by screening tests, performed with fraction I of Cohn. In some preparations esterase-activity was demonstrable on the substrates benzoyl-arginyl-ethylester (BAEE) and on glycerol tributyrate. There was, however, no correlation between these esterase-activities and the AHF-activity in the preparations. Moreover, the stability of the esterase at 4° C was much greater than the stability of the AHF-activity. We therefore assume that the AHF has no enzymatic activity on these substrates under the conditions of the tests. Attempts to activate AHF by calcium, magnesium or serum, were unsuccessful. Both esterase-activities were also present in fraction I of Cohn prepared from normal human citrated or resin plasma. The esterase-activity on BAEE is removed from resin plasma by adsorption to barium sulphate. The esterase-activity on glycerol tributyrate is not adsorbed.


1979 ◽  
Vol 42 (04) ◽  
pp. 1230-1239 ◽  
Author(s):  
I M Nilsson ◽  
T B L Kirkwood ◽  
T W Barrowcliffe

SummaryThe recovery and half-life of VIII: C in the plasma of severely haemophilic patients was measured by one-stage and two-stage assays after injection of two Factor VIII concentrates (Hemofil, Hyland and Fraction I-O, Kabi). Plasma volumes were measured with an Evans� Blue technique, and both concentrates and post-infusion samples were measured against the same plasma standard.There was a highly significant difference in recoveries estimated by the two assay methods. The one-stage assays gave the most consistent results, in that the average recovery was 100%, whereas the two-stage assays gave only about 80% of the value expected from in vitro assays. There was no difference in recoveries between the two concentrates.The two-stage assays gave a slightly shorter half-life than the one-stage assays, and the half-life of Hemofil was also shorter than that of Fraction I-O.


1975 ◽  
Author(s):  
J. D. Olson ◽  
D. N. Fass ◽  
W. J. Brockway ◽  
E. J. W. Bowie ◽  
K. G. Mann

A component required for the ristocetin-induced aggregation of platelets was isolated in yields of 20-30% from pooled human and porcine plasma by cryoethanol concentration of the RWF, after removal of the vitamin K dependent coagulation factors on barium citrate. The concentrate was further purified using gel filtration (4% agarose) and ion exchange (DEAE-cellulose) chromatography. Sodium dodecyl sulfate polyacrylamide gels of the isolated factor indicated an apparent molecular weight of greater than 500,000. After reduction of RWF with mercaptoethanol, a single band is resolved with an apparent molecular weight of 230,000. The purified component had no factor VIII procoagulant activity and did not compete for the activity of naturally occurring factor VIII inhibitor (human). Antisera raised in rabbits directed against the purified component inhibited the RWF activity but not the factor VIII procoagulant activity of plasma. Amino acid analysis indicated the presence of all normal amino acids and failed to detect any amino sugar. Analysis of lipid revealed a significant amount of lipid composed of mono, di and triglycerides, cholesterol, cholesterol esters and free fatty acid, with small portions of phospholipids.


1981 ◽  
Author(s):  
D L Aronson ◽  
J Bagley

The in vitro correction of the prolonged APTT of hemophilic plasma has been ascribed to an uncharacterized entity “Factor VIII Bypassing Activity.” Such products also correct the prolonged APTT plasma deficient in Factor IX, Factor X and Factor XII, but not of Factor V deficient plasma. Correction of the APTT in Factor VIII deficient plasma by early stage coagulants such as Factor XIIa, Kallikrein and Factor IXa is minimal. These results indicate that this in vitro activity acts at the level of either the activation of Factor X or the activation of prothrombin.A coagulant has been prepared from serum by barium precipitation, heparin-agarose, DEAE cellulose and high pressure liquid chromatography (HPLC). The in vitro coagulant properties are similar to “activated” prothrombin complex (Autoplex) and the biologic and chemical properties are identical to activated Factor X.Infusion of the partially purified serum coagulant into normal dogs was well tolerated and, in contrast to Factor IX concentrates, gave no signs of DIC. Infusion into bleeding hemophilic dogs had no hemostatic effect. It is concluded that a major portion of the in vitro potency of activated prothrombin concentrates is due to activated Factor X, a material which when infused has no in vivo hemostatic effect.Acknowledgments - The authors gratefully acknowledge the studies of Dr. Henry Kingdon in hemophilic dogs.


1986 ◽  
Vol 60 (6) ◽  
pp. 1917-1922 ◽  
Author(s):  
M. Andrew ◽  
C. Carter ◽  
H. O'Brodovich ◽  
G. Heigenhauser

Components of the factor VIII complex increase and activation of the fibrinolytic system occur during exercise. The relation between the duration and intensity of exercise and the relative changes in the VIII complex and fibrinolytic system have not been previously examined. Five healthy male subjects were exercised with three protocols: a graded progressive exercise test to exhaustion on a cycle ergometer with 50-W increments every 4 min, steady-state exercise, 15 min at 5 and 125 W each, and an acute 30-s maximal exercise test on a cycle ergometer. Venous blood samples were drawn at base line, during the last 30 s of each power output in the graded exercise, at 5-min intervals for the steady-state exercise, and for up to 1 h after completion of exercise in all three protocols. At the maximum exercise intensities, increases in plasma lactate concentration ([La]), O2 uptake, and [H+] were observed. Components of the VIII complex [VIII procoagulant, VIII procoagulant antigen, VIII-related antigen (VIIIR:Ag), VIII ristocetin cofactor activity] abruptly rose at only the highest work intensities, whereas the whole blood clot lysis time began to gradually shorten much earlier at low work intensities. There were no qualitative changes in the factor VIIIR:Ag on crossed immunoelectrophoresis nor was there evidence of thrombin generation as determined by fibrinopeptide A generation. We conclude that during exercise the changes observed in the coagulation and fibrinolytic systems are related to the intensity of the exercise, which is reflected by increases in plasma [La] and [H+], and that the fibrinolytic system is activated before the changes in the VIII complex are observed.


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