Skip to main content
NCBI home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1976 Apr;57(4):955–963. doi: 10.1172/JCI108372

Leukocyte-platelet interaction. Release of hydrogen peroxide by granulocytes as a modulator of platelet reactions.

P H Levine, R S Weinger, J Simon, K L Scoon, N I Krinsky
PMCID: PMC436739  PMID: 947961

Abstract

Because of the many potent biological capabilities of the blood granulocytes, and their contact with platelets in various physiologic and pathologic states, a possible interaction between granulocytes and platelets was investigated. Platelets were purified by gel filtration and via a dialysis membrane were separated from suspensions of autologous granulocytes prepared by dextran sedimentation and resuspended in modified Tyrode's buffer. After 20 min at 37 degrees C platelet aggregation was shown to be diminished by such exposure, as compared to the aggregation of platelets incubated with dialysates of buffer only. When granulocytes were stimulated by the addition of 1.1-muM latex spheres as target particles for phagocytes, the dialysate of these cells exhibited greatly enhanced platelet-inhibitory properties. The addition of catalase to the platelets abolished the effect of exposing these cells to the dialysate of resting granulocytes and markedly inhibited the effect of exposing the platelets to the dialysate of phagocytosing granulocytes. Catalase treated with 3-amino-1,2,4-triazole had no platelet-protective capacity. Purified suspensions of lymphocytes released no platelet-inhibitory principle under these experimental conditions. Hydrogen peroxide in the dialysate of granulocytes was measured directly with an assay involving an H2O2-induced decrease in the fluorescence of scopoletin catalyzed by horseradish peroxidase. The dialysate of phagocytosing granulocytes contained 0.86 +/- 0.55 nmol H2O2/2.5 X 10(7) granulocytes when sampled at 20 min. By an alternate measurement technique in which scopoletin and horseradish peroxidase were present in the dialysate from time zero, the mean amount of H2O2 in the dialysate reached 4.0 +/- 1.3 nmol/2.5 x 10(7) granulocytes at 20 min. This discrepancy suggested the consumption of H2O2, possibly mediated by the granulocytes themselves. This possibility was investigated by the addition of exogenous H2O2 to the test system. Both granulocytes and platelets enhanced the disappearance of H2O2 from the dialysate, and the amount consumed was proportional to the amount of H2O2 added to the system. Glucose oxidase at 12 M U/ml plus glucose in excess resulted in the production of H2O2 at a rate and final amount comparable to that produced by phagocytosing granulocytes. This mixture, when substituted for phagocytosing granulocytes in the standard dialysis membrane experiment, induced an inhibition of platelet aggregation similar to that caused by the granulocytes. The observation that the release of H2O2 by the blood granulocyte influences platelet function suggests a potential role for the granulocyte in the regulation of hemostasis or thrombosis.

Full text

PDF
955

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. ANDREAE W. A. A sensitive method for the estimation of hydrogen peroxide in biological materials. Nature. 1955 May 14;175(4463):859–860. doi: 10.1038/175859a0. [DOI] [PubMed] [Google Scholar]
  2. BORN G. V. Aggregation of blood platelets by adenosine diphosphate and its reversal. Nature. 1962 Jun 9;194:927–929. doi: 10.1038/194927b0. [DOI] [PubMed] [Google Scholar]
  3. Babior B. M., Kipnes R. S., Curnutte J. T. Biological defense mechanisms. The production by leukocytes of superoxide, a potential bactericidal agent. J Clin Invest. 1973 Mar;52(3):741–744. doi: 10.1172/JCI107236. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Baehner R. L., Gilman N., Karnovsky M. L. Respiration and glucose oxidation in human and guinea pig leukocytes: comparative studies. J Clin Invest. 1970 Apr;49(4):692–700. doi: 10.1172/JCI106281. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Baehner R. L., Nathan D. G., Castle W. B. Oxidant injury of caucasian glucose-6-phosphate dehydrogenase-deficient red blood cells by phagocytosing leukocytes during infection. J Clin Invest. 1971 Dec;50(12):2466–2473. doi: 10.1172/JCI106747. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Born G. V. Current ideas on the mechanism of platelet aggregation. Ann N Y Acad Sci. 1972 Oct 27;201:4–12. doi: 10.1111/j.1749-6632.1972.tb16283.x. [DOI] [PubMed] [Google Scholar]
  7. CAGAN R. H., KARNOVSKY M. L. ENZYMATIC BASIS OF THE RESPIRATORY STIMULATION DURING PHAGOCYTOSIS. Nature. 1964 Oct 17;204:255–257. doi: 10.1038/204255a0. [DOI] [PubMed] [Google Scholar]
  8. COHEN G., HOCHSTEIN P. GLUTATHIONE PEROXIDASE: THE PRIMARY AGENT FOR THE ELIMINATION OF HYDROGEN PEROXIDE IN ERYTHROCYTES. Biochemistry. 1963 Nov-Dec;2:1420–1428. doi: 10.1021/bi00906a038. [DOI] [PubMed] [Google Scholar]
  9. Canoso R. T., Rodvien R., Scoon K., Levine P. H. Hydrogen peroxide and platelet function. Blood. 1974 May;43(5):645–656. [PubMed] [Google Scholar]
  10. HENRY R. L. LEUKOCYTES AND THROMBOSIS. Thromb Diath Haemorrh. 1965 Mar 15;13:35–46. [PubMed] [Google Scholar]
  11. Hamberg M., Svensson J., Samuelsson B. Prostaglandin endoperoxides. A new concept concerning the mode of action and release of prostaglandins. Proc Natl Acad Sci U S A. 1974 Oct;71(10):3824–3828. doi: 10.1073/pnas.71.10.3824. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Harrison M. J., Emmons P. R., Mitchell J. R. The effect of white cells on platelet aggregation. Thromb Diath Haemorrh. 1966 Jul 31;16(1):105–121. [PubMed] [Google Scholar]
  13. Holmes B., Page A. R., Good R. A. Studies of the metabolic activity of leukocytes from patients with a genetic abnormality of phagocytic function. J Clin Invest. 1967 Sep;46(9):1422–1432. doi: 10.1172/JCI105634. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Jorgensen L., Rowsell H. C., Hovig T., Mustard J. F. Resolution and organization of platelet-rich mural thrombi in carotid arteries of swine. Am J Pathol. 1967 Nov;51(5):681–719. [PMC free article] [PubMed] [Google Scholar]
  15. MARGOLIASH E., NOVOGRODSKY A., SCHEJTER A. Irreversible reaction of 3-amino-1:2:4-triazole and related inhibitors with the protein of catalase. Biochem J. 1960 Feb;74:339–348. doi: 10.1042/bj0740339. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. MUSTARD J. F., HEGARDT B., ROWSELL H. C., MACMILLAN R. L. EFFECT OF ADENOSINE NUCLEOTIDES ON PLATELET AGGREGATION AND CLOTTING TIME. J Lab Clin Med. 1964 Oct;64:548–559. [PubMed] [Google Scholar]
  17. Mustard J. F., Kinlough-Rathbone R. L., Jenkins C. S., Packham M. A. Modification of platelet function. Ann N Y Acad Sci. 1972 Oct 27;201:343–359. doi: 10.1111/j.1749-6632.1972.tb16310.x. [DOI] [PubMed] [Google Scholar]
  18. Mustard J. F., Packham M. A. Thromboembolism: a manifestation of the response of blood to injury. Circulation. 1970 Jul;42(1):1–21. doi: 10.1161/01.cir.42.1.1. [DOI] [PubMed] [Google Scholar]
  19. Paul B., Sbarra A. J. The role of the phagocyte in host-parasite interactions. 13. The direct quantitative estimation of H2O2 in phagocytizing cells. Biochim Biophys Acta. 1968 Feb 1;156(1):168–178. doi: 10.1016/0304-4165(68)90116-5. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. SILVER M. D., STEHBENS W. E., SILVER M. M. PLATELET REACTION TO ADENOSINE DIPHOSPHATE IN VIVO. Nature. 1965 Jan 2;205:91–92. doi: 10.1038/205091a0. [DOI] [PubMed] [Google Scholar]
  22. Silver M. J., Smith J. B., Ingerman C., Kocsis J. J. Arachidonic acid-induced human platelet aggregation and prostaglandin formation. Prostaglandins. 1973 Dec;4(6):863–875. doi: 10.1016/0090-6980(73)90121-4. [DOI] [PubMed] [Google Scholar]
  23. Stemerman M. B. Vascular intimal components: precursors of thrombosis. Prog Hemost Thromb. 1974;2(0):1–47. [PubMed] [Google Scholar]
  24. Stocks J., Dormandy T. L. The autoxidation of human red cell lipids induced by hydrogen peroxide. Br J Haematol. 1971 Jan;20(1):95–111. doi: 10.1111/j.1365-2141.1971.tb00790.x. [DOI] [PubMed] [Google Scholar]
  25. Tangen O., Berman H. J., Marfey P. Gel filtration. A new technique for separation of blood platelets from plasma. Thromb Diath Haemorrh. 1971 Jun 30;25(2):268–278. [PubMed] [Google Scholar]
  26. Walsh P. N. Platelet coagulant activities: evidence for multiple different functions of platelets in intrinsic coagulation. Ser Haematol. 1973;6(4):579–592. [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES