The Coagulation Defect in Hemophilia. The Effect in Hemophilia of Intramuscular Administration. Of a Globulin Substance Derived From Normal Human Plasma 1

Abstract

The recent investigations of Patek and Stetson (1) 2 indicate that the defect in coagulation of blood in hemophilia resides in the plasma rather than in the platelets. Patek and Taylor (2, 3) 2 isolated a substance from citrated normal cellularfree plasma, by isoelectric precipitation, with which was associated the clot-promoting factor of normal plasma. This substance was effective, both in vitro and after intravenous injection, in reducing the coagulation time of hemophilic blood. By the same method, a similar substance in approximately the same quantity was obtained from citrated hemophilic plasma, but its clotaccelerating power on hemophilic blood was much less marked. In the absence of definite knowledge concerning the nature of the effective material it was called "globulin substance " which is the nomenclature that will be retained in this communication. The material was thermolabile, insoluble in water, partially soluble in physiological saline solution, and non-diffusible. It was precipitated in optimal amounts between pH 5.9 and 6.4. The material passed readily through a Berkefeld filter without loss of potency and was almost completely inactivated by small excesses of alkali (3). Bendien and Creveld (4) have isolated a substance, by dilution and acidification, from normal serum which they injected intramuscularly and intravenously and report favorable results.Patek and Taylor's (3) studies imply that in some respects hemophilia is a deficiency disease in which certain factors present in normal cellular-free plasma are either reduced or modified. The present communication reports a study of the effects of intramuscular administration of 1 The expenses of this research were defrayed in part by a gift to Harvard University from Smith, Kline and French Laboratories of Philadelphia.2 Review of literature here.globulin substance on the blood coagulation time of hemophilic subjects, and on the content of a clot-promoting factor in the blood following such administration. METHODSCoagulation time. In general the standard procedure followed was the same as that used in previous studies (1,3) When test substances were to be assayed they were pipetted into the test tubes prior to the addition of the blood. Further mixing was not found necessary.Immediately after the addition of whole blood, duplicate tubes were placed in a water bath at 37.50 C.

The Coagulation Defect in Hemophilia: The Clot Promoting Activity in Hemophilia of Berkefelded Normal Human Plasma Free From Fibrinogen and Prothrombin 1

“…A comparison of these data with those previously published for multiple injections of unmodified normal human plasma (10) shows a close correspondence. The maintenance of a lowered coagulation time by repeated injections of unmodified plasma and preparations of plasma free from prothrombin and fibrinogen is in sharp contrast to the refractory phase reported as accompanying multiple injections of acid-precipitated " globulin substance " (3).…”

The Coagulation Defect in Hemophilia: Studies of the Clot Promoting Activity Associated With Plasma Euglobulin in Hemophilia 1

“…During the past two years a study of the nature of " globulin substance " prepared by dilution and acid precipitation of plasma has been made (3,4,5). It was determined that the protein precipitates obtained between pH 5.5 and 6.0 are associated with the largest proportion of clot-promoting activity.…”

Boston city hospital and the thorndike memorial laboratory: the birth of modern haematology

“…Advances in the treatment of haemophilia with antihaemophilic factor began with pioneering work by Patek and Taylor (1937) when they demonstrated that this clot-promoting substance for haemophilic blood found in normal plasma was a protein ('globulin precipitate'). Pohle and Taylor (1937) first demonstrated that intramuscular use of this globulin substance was effective in correcting the clotting time in patients with haemophilia, but also demonstrated the refractory state induced by subsequent injections. The Thorndike also participated in the beginnings of the modern understanding of disseminated intravascular coagulation when Lozner and Taylor (1942) reported the effects of artificial surfaces on the initiation of coagulation and Tagnon et al (1942) studied the fibrinolytic activity of plasma.…”