Hematological Studies during Endotoxin Infusion in Rabbits

Summary

Endotoxin was infused over 14 hrs into rabbits at a rate of 30-50 μg/kg/hr. During the infusion of endotoxin the following parameters were observed:

a) The leukocytes declined in the early phase of the experiment but increased after 2 hrs despite continuation of the infusion.

b) Hemolysis in a range of 130-180 mg% plasma hemoglobin was present in animals at the time glomerular fibrin deposition was observed.

c) Platelets declined steadily until the end of the infusion.

d) Factor V declined during the infusion and reached levels below 10% of baseline (normal) levels.

e) The thrombelastogram revealed an increase in the r value and a decrease in m_ε.

f) Fibrinogen

g) plasminogen and

h) streptokinase inhibitor decreased initially reaching a plateau after 6 hrs.

i) Plasmin inhibitor and

j) urokinase inhibitor did not change during the period of observation.

k) Fibrin split products were demonstrated in 4 out of 7 serum samples taken 14 hrs following the initiation of the infusion.

I) C’activity decreased during infusion of endotoxin.

^* NIH Research Trainee in Reproductive Physiology (present address: Department of Obstetrics and Gynecology, Heidelberg University, 69 Heidelberg, Germany).

^** Career Scientist of the Health Research Council of the City of New York (I-297).

• References

• 1 Beller F.K, Graeff H. Deposition of glomerular fibrin in the rabbit after infusion with endotoxin. Nature 1967; 215: 295
• 2 Beller F.K, Graeff H, Gorstein F. Pathomorphical, pathophysiological events during the continous infusion of endotoxin in rabbits. Amer. J. Obstet. Gynec, (In press.)
• 3 Rodriguez-Erdmann F. Über eine einfache Methode zur wiederholten Blutgewinnung beim Kaninchen. Pfluegers Arch. ges. Physiol 1959; 269: 306
• 4 Hartert H. Die Thrombelastographie, eine Methode zur physikalischen Analyse des Blutgerinnungsvorganges. Z. ges. exp. Med 1951; 117: 189
• 5 Beller F.K, Koch F, Mamm E. Thrombelastographische Untersuchungen bei isolierten Störungen der Vor- und I. Phase der Gerinnung. Blut 1956; 2: 112
• 6 Ratnoff O.D, Menzie C. Estimation of fibrinogen in small samples of plasma. In: Blood coagulation, hemorrhage and thrombosis. Eds. Tocantins L.M, Kazal L.A. Grune and Stratton; New York, London: 1964: p. 224
• 7 Jürgens J, Beller F.K. Klinische Methoden der Blutgerinnungsanalyse. Thieme; Stuttgart: 1959: p. 207
• 8 Johnson A.J, Tillett W.S. The lysis in rabbits of intravascular blood clots by the streptococcal fibrinolytic system. J. exp. Med 1952; 95: 449
• 9 Milstone H. A factor in normal human blood which participates in streptococcal fibrinolysis. J. Immunol 1941; 42: 109
• 10 Graeff H, Beller FK. Plasminogen assays using fibrin as a substrate. In: Theory and Methods of Thrombosis and Hemorrhage Eds. Bang N, Beller FK, Deutsch E, Mammen E.E. Thieme, Stuttgart: New York Academic press; (In press.)
• 11 Johnson A.J, Tse A.O, Newman J. Fibrinolytic determination of plasmin inhibitor in purified systems or biological fluids. In: Blood coagulation, hemorrhage and thrombosis. Eds. Tocantins L.M, Kazal L.A. Grune and Stratton; New York, London: 1964: 481-482
• 12 Johnson A.J, Tse A.O, Newman J. Fibrinolytic Estimation of Streptokinase (SK) inhibitor and antibody, and Urokinase (UK) inhibitor in purified systems or biological fluids. In: Blood coagulation, hemorrhage and thrombosis. Eds. Tocantins L.M, Kazal L.A. Grune and Stratton; New York, London: 1964: 479-481
• 13 Bennett N.B. A method for the quantitative assay of inhibitor of plasminogen activation in human serum. Thrombos. Diathes. haemorrh. (Stuttg.) 1967; 17: 12
• 14 Mancini S, Vaerman J.P, Garbonara O, Heremans J.F. A single radial diffusion method for the immunological quantitation of proteins. In: Proc. XIth Colloquium Proteins of the biolog. fluids. Ed. Peters H. Elzevier; Amsterdam: 1964: p. 370
• 15 Beller F.K, Maki M. Properties of fibrinogenolysis and fibrinolysis products in immune assays. Thrombos. Diathes. haemorrh. (Stuttg.) 1967; 18: 114
• 16 Mayer M.M. Complement Complement fixation. In: Kabat Mayer's Experimental Immunochemistry II.. Thomas; Springfield (Ill.) Ch. C.: 1961: 133-240
• 17 Apitz K. A study of the generalized Shwartzman phenomenon. J. Immunol 1935; 29: 255
• 18 Shwartzman G, Gerber J.E. Hemorrhagic manifestations of bacterial and virus infections: Experimental studies and pathological interpretations. Ann. N. Y. Acad. Sci 1948; 49: 627
• 19 McKay D.G, Shapiro S.S. Alterations in the blood coagulation system induced by bacterial endotoxin. I. in vivo (Generalized Shwartzman Reaction). J. exp. Med 1958; 107: 353
• 20 Kleinmaier H, Georgen K, Lasch H.G, Krecke H.-J, Bohle A. Untersuchungen zur Frage der Gerirmungsstörung beim Sanarelli-Shwartzman-Phänomen des Kaninchens. Z. ges. exp. Med 1959; 132: 275
• 21 Horowitz H.J, des Prez R.M, Hook E.W. Effects of bacterial endotoxin on rabbit platelets. II. Enhancement of platelet 3 activity in vitro and in vivo. J. exp. Med 1962; 116: 619
• 22 McKay D.G, Müller-Berghaus G. Hageman Factor and the Generalized Shwartzman Reaction. Fed. Proc 1968; 27: 1229
• 23 McKay D.G, Margaretten W, Csavossy J. An electron mecroscope study of the effects of bacterial endotoxin on the blood - vascular system. Lab. Invest 1966; 15: 1815
• 24 erghaus Müller-B.G, Krecke H.J, Lasch H.G. Thrombelastographische Untersuchungen zum Gerinnselaufbau beim Sanarelli-Shwartzman-Phänomen. Klin. Wschr 1963; 41: 216
• 25 Schimpf Kl, Gibelins A. Die physiologischen Inhibitoren der Blutgerinnung bei pathologisch gesteigerter Gerinnbarkeit in vivo. Klin. Wschr 1964; 42: 431
• 26 Lasch H.G, Rodriguez-Erdmann F. Kl. Schimpf: Antithrombin III und Anti-Blutthrombokinase bei experimenteller Verbrauchkoagulopathie. (Am Beispiel des Sanarelli-Shwartzman-Phänomens). Klin. Wschr 1967; 39: 645
• 27 Graeff H, Kuhn W. Beeinflussung der Vorphase der Gerinnung durch Dextran 150. II. Mitt. : In Vivo Studien am Kaninchen. Klin. Wschr 1966; 44: 646
• 28 Lee L. Reticuloendothelial clearance of circulating fibrin in the pathogenesis of the generalized Shwartzman reaction. J. exp. Med 1962; 115: 1065
• 29 Margaretten W, Zunker H.O, McKay D.G. Production of the generalized Shwartzman reaction in pregnant rats by intravenous infusion of thrombin. Lab. Invest 1964; 13: 552
• 30 Beller F.K, Mitchell P.S, Gorstein F. Fibrin deposition in the rabbit kidney produced by protease inhibitors. Thrombos. Diathes. haemorrh. (Stuttg.) 1967; 17: 427
• 31 Graeff H, Kuhn W, Bleyl U. Verbrauchskoagulopathie und Lysocoagulopathie bei menschlichen Äquivalenten des Sanarelli-Shwartzman-Phänomens (generalisertes Shwartzman -Phänomen). Thrombos. Diathes. haemorrh. (Stuttg.) 1967; 17: 144
• 32 Merskey C, Johnson A.J, Kleiner G.J, Wohl H. The defibrination syndrome. Clinical features and laboratory diagnosis. Brit. J. Haemat 1967; 13: 528
• 33 Merskey C, Johnson A.J. Diagnosis and treatment of intravascular coagulation. Thrombos. Diathes. haemorrh. (Stuttg.) 1966; 20: 555 Suppl
• 34 Stetson C.A. Studies on the mechanism of the Shwartzman phenomenon. Certain factors involved in the production of the local haemorrhagic necrosis. J. exp. Med 1951; 93: 489
• 35 Thomas L, Good R.A. Studies on the generalized Shwartzman reaction. I. General observations concerning the phenomenon. J. exp. Med 1952; 96: 605
• 36 McKay D.G, Margaretten W, Phillips L.L. The role of leukocyte in the Generalized Shwartzman Reaction. Lab. Invest 1967; 16: 511
• 37 Gilbert V.E, Braude A.I. Reduction of serum complement in rabbits after injection of endotoxin. J. exp. Med 1962; 116: 477
• 38 Pearlman D.S, Sauers J.B, Talmage P.W. The effect of adjuvant amounts of endotoxins on the serum hemolytic complement activity in rabbits. J. Immun 1963; 91: 748
• 39 Gewurz H, Bladen H.A, Mergenhagen S.E. Interactions of the complement (C’) system with endotoxic lipopolysaccharide (LPS). Proc. Soc. exp. Biol. (N. Y.) 1967; 26: 584
• 40 Ho M, Kass E.H. Hemolytic anemia in rabbits following injection of bacterial endotoxin. Proc. Soc. exp. Biol. (N. Y.) 1958; 97: 505
• 41 Krecke H.J, Bohle A, Lasch H.G, Becker B. Klinisch-pathol. und experiment. Untersuchungen zur bilateralen Nierenrindennekrose. In: Aktuelle Probleme der Nephrologie. Ed. Krueck F. Springer; Berlin, Heidelberg: 1966: p. 640
• 42 Brain M.G, Hourihane D.O. B. Microangiopathic anemia: The occurrence of hemolysis in experimentally produced vascular disease. Brit. J. Haemat 1967; 13: 135
• 43 Regoeczi E, Rubenberg M.C, Brain M.C. Intravascular haemolysis and disseminated intravascular coagulation. Lancet 1967; 7490: 601