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. 1967 Aug 1;126(2):235–250. doi: 10.1084/jem.126.2.235

AN ULTRASTRUCTURAL STUDY OF THE MECHANISMS OF PLATELET-ENDOTOXIN INTERACTION

Arthur R Spielvogel 1
PMCID: PMC2138316  PMID: 4961486

Abstract

Electron microscopy has confirmed previous studies and has provided much new information on the mechanism of endotoxin-platelet interaction. The Boivin lipopolysaccharide preparation is particulate, and on electron microscope examination appears as a three-layered structure, morphologically similar to bacterial cell wall. In vitro and in vivo experiments have demonstrated that these endotoxin particles adhere to platelets. In some species, particularly the rabbit, this is associated with loss of platelet contents, due to lysosomal and cell membrane lysis, resulting in platelet sphering and apparent cell death. Serial observation of degranulating platelets and metabolic studies indicate that although some platelet engulfment of endotoxin occurs, degranulation is not dependent upon phagocytosis. Several observations suggest that these endotoxin effects are mediated through immune mechanisms: (1) Inactivation of complement in the suspending plasma by heating to 56°C, anticoagulation with EDTA, a reaction temperature of 5°C, ammonium hydroxide incubation, and adsorption with either zymosan or washed antigen-antibody complexes, inhibits both endotoxin adherence and platelet degranulation. (2) The reaction requires a plasma cofactor, possibly antibody, which can be adsorbed out by endotoxin. (3) Endotoxin adheres selectively to nonprimate platelets and primate red cells, a pattern conforming to immune adherence, a phenomenon requiring antigen, antibody, and complement. It is suggested that endotoxin-induced platelet damage is dependent upon the intimate contact provided by immune adherence. We have not established whether degranulation is an endotoxin or complement effect. The species variation in susceptibility to endotoxin also merits further investigation.

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Selected References

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  1. BENNETT I. L., Jr, BEESON P. B. The properties and biologic effects of bacterial pyrogens. Medicine (Baltimore) 1950 Dec;29(4):365–400. doi: 10.1097/00005792-195012000-00003. [DOI] [PubMed] [Google Scholar]
  2. BRAUDE A. I., CAREY F. J., ZALESKY M. Studies with radioactive endotoxin. II. Correlation of physiologic effects with distribution of radioactivity in rabbits injected with radioactive sodium chromate. J Clin Invest. 1955 Jun;34(6):858–866. doi: 10.1172/JCI103141. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. CASTALDI P. A., FIRKIN B. G., BLACKWELL P. M., CLIFFORD K. I. An electron microscopic study of the changes in platelets during viscous metamorphosis. Blood. 1962 Nov;20:566–580. [PubMed] [Google Scholar]
  4. COHEN P., BRAUNWALD J., GARDNER F. H. DESTRUCTION OF CANINE AND RABBIT PLATELETS FOLLOWING INTRAVENOUS ADMINISTRATION OF CARBON PARTICLES OR ENDOTOXIN. J Lab Clin Med. 1965 Aug;66:263–271. [PubMed] [Google Scholar]
  5. COHN Z. A., MORSE S. I. Functional and metabolic properties of polymorphonuclear leucocytes. I. Observations on the requirements and consequences of particle ingestion. J Exp Med. 1960 May 1;111:667–687. doi: 10.1084/jem.111.5.667. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cohen P., Gardner F. H. Thrombocytopenia as a laboratory sign and complication of gram-negative bacteremic infection. Arch Intern Med. 1966 Jan;117(1):113–124. [PubMed] [Google Scholar]
  7. DAVIS R. B., MEEKER W. R., McQUARRIE D. G. Immediate effects of intravenous endotoxin on serotonin concentrations and blood platelets. Circ Res. 1960 Jan;8:234–239. doi: 10.1161/01.res.8.1.234. [DOI] [PubMed] [Google Scholar]
  8. DES PREZ R. M., HOROWITZ H. I., HOOK E. W. Effects of bacterial endotoxin on rabbit platelets. I. Platelet aggregation and release of platelet factors in vitro. J Exp Med. 1961 Dec 1;114:857–874. doi: 10.1084/jem.114.6.857. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Davis R. B. Electron microscopic changes in blood platelets induced by bacterial lipopolysaccharide. Exp Mol Pathol. 1966 Dec;5(6):559–574. doi: 10.1016/0014-4800(66)90046-3. [DOI] [PubMed] [Google Scholar]
  10. Des Prez R. M., Bryant R. E. Effects of bacterial endotoxin on rabbit platelets. IV. The divalent ion requirements of endotoxin-induced and immunologically induced platelet injury. J Exp Med. 1966 Nov 1;124(5):971–982. doi: 10.1084/jem.124.5.971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. GILBERT V. E., BRAUDE A. I. Reduction of serum complement in rabbits after injection of endotoxin. J Exp Med. 1962 Oct 1;116:477–490. doi: 10.1084/jem.116.4.477. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. GOCKE D. J., OSLER A. G. IN VITRO DAMAGE OF RABBIT PLATELETS BY AN UNRELATED ANTIGEN-ANTIBODY REACTION. I. GENERAL CHARACTERISTICS OF THE REACTION. J Immunol. 1965 Feb;94:236–246. [PubMed] [Google Scholar]
  13. HERRING W. B., HERION J. C., WALKER R. I., PALMER J. G. Distribution and clearance of circulating endotoxin. J Clin Invest. 1963 Jan;42:79–87. doi: 10.1172/JCI104698. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. HINSHAW L. B., JORDAN M. M., VICK J. A. Mechanism of histamine release in endotoxin shock. Am J Physiol. 1961 May;200:987–989. doi: 10.1152/ajplegacy.1961.200.5.987. [DOI] [PubMed] [Google Scholar]
  15. HOROWITZ H. I., DES PREZ R. M., HOOK E. W. Effects of bacterial endotoxin on rabbit platelets. II. Enhancement of platelet factor 3 activity in vitro and in vivo. J Exp Med. 1962 Nov 1;116:619–633. doi: 10.1084/jem.116.5.619. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. HOVIG T. AGGREGATION OF RABBIT BLOOD PLATELETS PRODUCED IN VITRO BY SALINE "EXTRACT" OF TENDONS. Thromb Diath Haemorrh. 1963 Jul 15;143:248–263. [PubMed] [Google Scholar]
  17. HOVIG T. The ultrastructure of rabbit blood platelet aggregates. Thromb Diath Haemorrh. 1962 Dec 20;8:455–471. [PubMed] [Google Scholar]
  18. ISHIZAKA T., ISHIZAKA K. Biological activities of aggregated gamma globulin. I. Skin reactive and complement-fixing properties of heat denatured gamma globulin. Proc Soc Exp Biol Med. 1959 Aug-Sep;101:845–850. doi: 10.3181/00379727-101-25116. [DOI] [PubMed] [Google Scholar]
  19. LUFT J. H. Improvements in epoxy resin embedding methods. J Biophys Biochem Cytol. 1961 Feb;9:409–414. doi: 10.1083/jcb.9.2.409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. MICHAEL J. G., WHITBY J. L., LANDY M. Studies on natural antibodies to gram-negative bacteria. J Exp Med. 1962 Jan 1;115:131–146. doi: 10.1084/jem.115.1.131. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Movat H. Z., Weiser W. J., Glynn M. F., Mustard J. F. Platelet phagocytosis and aggregation. J Cell Biol. 1965 Dec;27(3):531–543. doi: 10.1083/jcb.27.3.531. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. NELSON D. S., NELSON R. A., Jr On the mechanism of immune-adherence. I. Differentiation from acid-adhesion of bacteria to erythrocytes. Yale J Biol Med. 1959 Feb;31(4):185–200. [PMC free article] [PubMed] [Google Scholar]
  23. NELSON R. A., Jr, NELSON D. S. On the mechanism of immune-adherence. II. Analogy to mixed aggregation of sensitized antigens in the presence of complement; immune-adherence with animal platelets. Yale J Biol Med. 1959 Feb;31(4):201–212. [PMC free article] [PubMed] [Google Scholar]
  24. PILLEMER L., SCHOENBERG M. D., BLUM L., WURZ L. Properdin system and immunity. II. Interaction of the properdin system with polysaccharides. Science. 1955 Sep 23;122(3169):545–549. doi: 10.1126/science.122.3169.545. [DOI] [PubMed] [Google Scholar]
  25. REAM V. J., DEYKIN D., GUREWICH V., WESSLER S. THE AGGREGATION OF HUMAN PLATELETS BY BACTERIAL ENDOTOXIN. J Lab Clin Med. 1965 Aug;66:245–252. [PubMed] [Google Scholar]
  26. REYNOLDS E. S. The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. J Cell Biol. 1963 Apr;17:208–212. doi: 10.1083/jcb.17.1.208. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. SBARRA A. J., KARNOVSKY M. L. The biochemical basis of phagocytosis. I. Metabolic changes during the ingestion of particles by polymorphonuclear leukocytes. J Biol Chem. 1959 Jun;234(6):1355–1362. [PubMed] [Google Scholar]
  28. SPAET T. H., CINTRON J., KROPATKIN M. A technique for determining whole blood clotting times in plastic tubes. J Lab Clin Med. 1959 Sep;54:467–470. [PubMed] [Google Scholar]
  29. STETSON C. A., Jr Studies on the mechanism of the Shwartzman phenomenon; certain factors involved in the production of the local hemorrhagic necrosis. J Exp Med. 1951 May;93(5):489–504. doi: 10.1084/jem.93.5.489. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Spink W. W., Vick J. A LABILE SERUM FACTOR IN EXPERIMENTAL ENDOTOXIN SHOCK: CROSS-TRANSFUSION STUDIES IN DOGS. J Exp Med. 1961 Sep 30;114(4):501–508. doi: 10.1084/jem.114.4.501. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. WATSON M. L. Staining of tissue sections for electron microscopy with heavy metals. J Biophys Biochem Cytol. 1958 Jul 25;4(4):475–478. doi: 10.1083/jcb.4.4.475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. WHITE J. G., KRIVIT W., VERNIER R. L. THE PLATELET-FIBRIN RELATIONSHIP IN HUMAN BLOOD CLOTS: AN ULTRASTRUCTURAL STUDY UTILIZING FERRITIN-CONJUGATED ANTI-HUMAN FIBRINOGEN ANTIBODY. Blood. 1965 Feb;25:241–257. [PubMed] [Google Scholar]
  33. ZUCKER-FRANKLIN D. ELECTRON MICROSCOPE STUDY OF THE DEGRANULATION OF POLYMORPHONUCLEAR LEUKOCYTES FOLLOWING TREATMENT WITH STREPTOLYSIN. Am J Pathol. 1965 Sep;47:419–433. [PMC free article] [PubMed] [Google Scholar]

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