The Blood Clotting Properties of Rabbit Peritoneal Leukocytes in Vitro^[*]

Summary

A systematic in vitro study has been carried out of the blood clotting properties of rabbit peritoneal leukocytes. Rabbit heterophils have been shown to possess weak but definite tissue thromboplastic activity. They also contain an anticoagulant activity which is active in the intrinsic clotting system, in the extrinsic clotting system with brain thromboplastin, and in clotting systems with Russell’s viper venom. When the thromboplastic activity of the white cell itself initiates clotting, the anticoagulant is much less effective and the procoagulant effect of the WBC suspension predominates. Rabbit heterophils do not bind plasma clotting factors, do not activative factor V, and, under our conditions, do not precipitate fibrinogen from normal plasma, from plasma exposed to traces of thrombin in vitro, or from plasma of rabbits given endotoxin.

The relation of these findings to the role of the granulocyte in the pathogenesis of the generalized Shwartzman reaction has been discussed.

• References

• 1 Thomas L, Good R. A. Studies on the generalized Shwartzman reaction. I. General observations concerning the phenomenon. J. exp. Med 96: 605 1952;
• 2 Hjort P. F, McGehee W, Rapaport S. I. Studies on the effect of granulocytes in the generalized Shwartzman reaction. Abstract, XII Congress of the International Society of Haematology, 1964, Stockholm, paper G: 1.
• 3 Thomas L, Smith R. T, von Korjf R. Studies on the generalized Shwartzman reaction. VII. The role of fibrinogen in the deposition of fibrinoid after combined injections of endotoxin and synthetic acidic polymer. J. exp. Med 102: 263 1955;
• 4 Horn R. G, Spicer S. S. Sulfated mucopolysaccharide and basic protein in certain granules of circulating heterophils of rabbits during endotoxin-induced leukocytosis. Amer. J. Path 44: 905 1964;
• 5 Fedorko M. E, Morse S. I. Isolation, characterization, and distribution of acid mucopolysaccharides in rabbit leucocytes. J. exp. Med 121: 39 1965;
• 6 Rapaport S. I, Schiffman S, Patch M. J, Ames S. B. The importance of activation of antihemophilic globulin and proaccelerin by traces of thrombin in the generation of intrinsic prothrombinase activity. Blood 21: 221 1963;
• 7 Husom O. A one stage method for the assay of platelet factor 3. Scand. J. clin. Lab. Invest 13: 609 1961;
• 8 Hjort P. F, Rapaport S. I, Owren P. A. A simple specific one-stage prothrombin assay using Russell’s viper venom in cephalin suspension. J. Lab. clin. Med 46: 89 1955;
• 9 Owren P. A. A quantitative one-stage method for the assay of prothrombin. Scand. J. clin. Lab. Invest 1: 81 1949;
• 10 Hirsch J. G. Phagocytin: A bacterial substance from polymorphonuclear leucocytes. J. exp. Med 103: 589 1956;
• 11 Richter M. N, MacDowell E. G. Studies on mouse leukemia. VII. The relation of cell death to the potency of inoculated cell suspensions. J. exp. Med 57: 1 1933;
• 12 Schreck R. Studies in vitro on the physiology of normal and of cancerous cells. I. The effect of high temperature and of moccasin venom on the viability of rabbit lymphocytes and polymorphonuclear leukocytes as determined by the method of unstained cell counts. Arch. Path 35: 857 1943;
• 13 Horn R. G, Spicer S. S. Sulfated mucopolysaccharide and basic protein in certain granules of rabbit leukocytes. Lab. Invest 13: 1 1964;
• 14 Cohn Z. A, Hirsch J. G. The isolation and properties of specific cytoplasmic granules of rabbit polymorphonuclear leucocytes. J. exp. Med 112: 983 1960;
• 15 Borchgrevink C. F, Pool J. G, Stormorken H. A new assay for Factor V (proaccelerin-accelerin) using Russell’s viper venom. J. Lab. clin. Med 55: 625 1960;
• 16 Aas K. Prokonvertin og konvertin. Thesis, Oslo : Akademisk Trykningssentral, 1952. 90 pp.
• 17 Schiffman S, Rapaport S. I, Patch M. J. The identification and synthesis of activated plasma thromboplastin component (PTC). Blood 22: 733 1963;
• 18 Hougie G. A simple assay method for Factor X (Stuart-Prower factor). Proc. Soc. exp. Biol. (N. Y.) 109: 754 1962;
• 19 Hjort P. Intermediate reactions in the coagulation of blood with tissue thromboplastin. Scand. J. clin. Lab. Invest 9 (Suppl. 27) 183 1957;
• 20 Gans H. Fibrinolytic properties of proteases derived from human, dog and rabbit leukocytes. Thrombos. Diathes. haemorrh. (Stuttg.) 10: 379 1964;
• 21 Markwardt F. Blutgerinnungshemmende Wirkstoffe aus blutsaugenden Tieren. Fischer; Jena: 1963: 122
• 22 Martin H, Roka L. Beeinflussung der Blutgerinnung durch Leukocyten. Klin. Wschr 29: 510 1951;
• 23 Eiseman G, Stafenini M. Thromboplastic activity of leukemic white cells. Proc. Soc. exp. Biol. (N. Y.) 86: 763 1954;
• 24 Kerby G. P. Release of enzyme from human leukocytes on damage by bacterial derivatives. Proc. Soc. exp. Biol. (N. Y.) 81: 381 1952;
• 25 Hénon M, Delaunay A, Bazin S. Modifications enzymatiques produites dans les leucocytes et le sérum de cobayes intoxiqués par une endotoxine bactérienne. Ann. Inst. Pasteur 107: 604 1964;
• 26 Baker W. G, Bang N. V, Nachman R. L, Raafat F, Horowitz H. I. Hypofibrinogenemic hemorrhage in acute myelogenous leukemia treated with heparin. Ann. intern. Med 61: 116 1964;
• 27 Kopp W. L. Demonstration of serum proteins on washed leukocytes by means of fluorescent antibodies. J. Lab. clin. Med 62: 18 1963;
• 28 Horn R. G, Spicer S. S. Sulfated mucopolysaccharide in fibrinoid glomerular occlusions of the generalized Shwartzman reaction. Amer. J. Path 46: 197 1965;
• 29 Graham Jr. R. C, Ebert R. H, Ratnoff O. D, Moses J. M. Pathogenesis of inflammation. II. In vivo observations of the inflammatory effects of activated Hageman factor and brady-kinin. J. exp. Med 121: 807 1965;