Isolation And Characterization Of Procoagulant Substances From Human Ascites

Abstract

The treatment of massive, medically intractable ascites by a peritoneovenous shunt (PVS) is associated with variably severe disseminated intravascular coagulation (DIC). Ascitic fluid obtained from cirrhotic patients at the time of placement of a PVS was found to shorten the partial thromboplastin time (PTT) of normal human platelet-poor plasma. This procoagulant activity which was found to reside in the cell-free fraction of ascitic fluid was heat stable and insensitive to pH change over a wide range. Chromatography on Bio-Rad Agarose 1.5 yielded one major high molecular weight component and several smaller fractions of lower molecular weight exhibiting procoagulant activity as determined by the PTT assay. The activity in the major fraction (80% total activity) coprecipitated with human fibrinogen but could be separated from fibrinogen by affinity chromatography using anti-human fibrinogen. This procoagulant did not hydrolyze chromogenic substrates S- 2222 and S-2238 and was not inhibited by diisopropylfluoro-phosphate (DFP). Purification of a minor procoagulant fraction by aluminum hydroxide (Al (OH)3) adsorption and chromatography on Sephadex G-200 yielded a fraction that clotted both citrated normal plasma (without the addition of calcium chloride) and purified human fibrinogen and induced platelet aggregation in citrated human platelet- rich plasma. Chromatography on DEAE cellulose yielded two peaks with procoagulant activity (PTT), one of which hydrolyzed S-2238 and was DFP-sensitive. These studies indicate that at least three distinct, clotting active substances are present in ascitic fluid some or all of which may be responsible for PVS-induced DIC. Further studies on purification and characterization of these ascitic procoagulants are in progress.