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. 1985 Nov;76(5):2020–2025. doi: 10.1172/JCI112203

Immunoglobulin G is a platelet alpha granule-secreted protein.

J N George, S Saucerman, S P Levine, L K Knieriem, D F Bainton
PMCID: PMC424269  PMID: 2932474

Abstract

It has been known for 27 yr that blood platelets contain IgG, yet its subcellular location and significance have never been clearly determined. In these studies, the location of IgG within human platelets was investigated by immunocytochemical techniques and by the response of platelet IgG to agents that cause platelet secretion. Using frozen thin-sections of platelets and an immunogold probe, IgG was located within the alpha-granules. Thrombin stimulation caused parallel secretion of platelet IgG and two known alpha-granule proteins, platelet factor 4 and beta-thromboglobulin, beginning at 0.02 U/ml and reaching 100% at 0.5 U/ml. Thrombin-induced secretion of all three proteins was inhibited by prostaglandin E1 and dibutyryl-cyclic AMP. Calcium ionophore A23187 also caused parallel secretion of all three proteins, whereas ADP caused virtually no secretion of any of the three. From these data and a review of the literature, we hypothesize that plasma IgG is taken up by megakaryocytes and delivered to the alpha-granules, where it is stored for later secretion by mature platelets.

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Selected References

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  1. Chiu H. C., Schick P. K., Colman R. W. Biosynthesis of factor V in isolated guinea pig megakaryocytes. J Clin Invest. 1985 Feb;75(2):339–346. doi: 10.1172/JCI111706. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Cines D. B., Schreiber A. D. Immune thrombocytopenia. Use of a Coombs antiglobulin test to detect IgG and C3 on platelets. N Engl J Med. 1979 Jan 18;300(3):106–111. doi: 10.1056/NEJM197901183000302. [DOI] [PubMed] [Google Scholar]
  3. Davey M. G., Lüscher E. F. Actions of thrombin and other coagulant and proteolytic enzymes on blood platelets. Nature. 1967 Dec 2;216(5118):857–858. doi: 10.1038/216857a0. [DOI] [PubMed] [Google Scholar]
  4. De Bruyn P. P. Structural substrates of bone marrow function. Semin Hematol. 1981 Jul;18(3):179–193. [PubMed] [Google Scholar]
  5. Dixon R., Rosse W., Ebbert L. Quantitative determination of antibody in idiopathic thrombocytopenic purpura. Correlation of serum and platelet-bound antibody with clinical response. N Engl J Med. 1975 Jan 30;292(5):230–236. doi: 10.1056/NEJM197501302920503. [DOI] [PubMed] [Google Scholar]
  6. Dvorak A. M., Klebanoff S. J., Henderson W. R., Monahan R. A., Pyne K., Galli S. J. Vesicular uptake of eosinophil peroxidase by guinea pig basophils and by cloned mouse mast cells and granule-containing lymphoid cells. Am J Pathol. 1985 Mar;118(3):425–438. [PMC free article] [PubMed] [Google Scholar]
  7. Files J. C., Malpass T. W., Yee E. K., Ritchie J. L., Harker L. A. Studies of human plate alpha-granule release in vivo. Blood. 1981 Sep;58(3):607–618. [PubMed] [Google Scholar]
  8. George J. N., Lyons R. M., Morgan R. K. Membrane changes associated with platelet activation. Exposure of actin on the platelet surface after thrombin-induced secretion. J Clin Invest. 1980 Jul;66(1):1–9. doi: 10.1172/JCI109821. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. George J. N., Nurden A. T., Phillips D. R. Molecular defects in interactions of platelets with the vessel wall. N Engl J Med. 1984 Oct 25;311(17):1084–1098. doi: 10.1056/NEJM198410253111705. [DOI] [PubMed] [Google Scholar]
  10. George J. N. Studies on platelet plasma membranes. IV. Quantitative analysis of platelet membrane glycoproteins by (125I)-diazotized diiodosulfanilic acid labeling and SDS-polyacrylamide gel electrophoresis. J Lab Clin Med. 1978 Sep;92(3):430–446. [PubMed] [Google Scholar]
  11. George J. N., Thoi L. L., Morgan R. K. Quantitative analysis of platelet membrane glycoproteins: effect of platelet washing procedures and isolation of platelet density subpopulations. Thromb Res. 1981 Jul 1;23(1-2):69–77. doi: 10.1016/0049-3848(81)90239-5. [DOI] [PubMed] [Google Scholar]
  12. Griffiths G., Brands R., Burke B., Louvard D., Warren G. Viral membrane proteins acquire galactose in trans Golgi cisternae during intracellular transport. J Cell Biol. 1982 Dec;95(3):781–792. doi: 10.1083/jcb.95.3.781. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gyotoku Y., Mori M., Nonaka Y., Nagata Y., Saito Y. Differences of immunoglobulin secreting cells in bone marrow from those in circulation: relevance to their development. Br J Haematol. 1985 Apr;59(4):647–657. doi: 10.1111/j.1365-2141.1985.tb07360.x. [DOI] [PubMed] [Google Scholar]
  14. Hymes K., Schur P. H., Karpatkin S. Heavy-chain subclass of round antiplatelet IgG in autoimmune thrombocytopenic purpura. Blood. 1980 Jul;56(1):84–87. [PubMed] [Google Scholar]
  15. Kelton J. G., Denomme G., Lucarelli A., Garvey J., Powers P., Carter C. Comparison of the measurement of surface or total platelet-associated IgG in the diagnosis of immune thrombocytopenia. Am J Hematol. 1985 Jan;18(1):1–5. doi: 10.1002/ajh.2830180102. [DOI] [PubMed] [Google Scholar]
  16. Kelton J. G., Steeves K. The amount of platelet-bound albumin parallels the amount of IgG on washed platelets from patients with immune thrombocytopenia. Blood. 1983 Oct;62(4):924–927. [PubMed] [Google Scholar]
  17. Kelton J. G. The measurement of platelet-bound immunoglobulins: an overview of the methods and the biological relevance of platelet-associated IgG. Prog Hematol. 1983;13:163–199. [PubMed] [Google Scholar]
  18. Kernoff L. M., Blake K. C., Shackleton D. Influence of the amount of platelet-bound IgG on platelet survival and site of sequestration in autoimmune thrombocytopenia. Blood. 1980 May;55(5):730–733. [PubMed] [Google Scholar]
  19. Kieffer N., Boizard B., Didry D., Wautier J. L., Nurden A. T. Immunochemical characterization of the platelet-specific alloantigen Leka: a comparative study with the PlA1 alloantigen. Blood. 1984 Dec;64(6):1212–1219. [PubMed] [Google Scholar]
  20. Leporrier M., Dighiero G., Auzemery M., Binet J. L. Detection and quantification of platelet-bound antibodies with immunoperoxidase. Br J Haematol. 1979 Aug;42(4):605–611. doi: 10.1111/j.1365-2141.1979.tb01173.x. [DOI] [PubMed] [Google Scholar]
  21. Leven R. M., Schick P. K., Budzynski A. Z. Fibrinogen biosynthesis in isolated guinea pig megakaryocytes. Blood. 1985 Feb;65(2):501–504. [PubMed] [Google Scholar]
  22. Levine S. P., Krentz L. S. Development of a radioimmunoassay for human platelet factor 4. Thromb Res. 1977 Nov;11(5):673–686. doi: 10.1016/0049-3848(77)90025-1. [DOI] [PubMed] [Google Scholar]
  23. Lichtman M. A., Chamberlain J. K., Simon W., Santillo P. A. Parasinusoidal location of megakaryocytes in marrow: a determinant of platelet release. Am J Hematol. 1978;4(4):303–312. doi: 10.1002/ajh.2830040402. [DOI] [PubMed] [Google Scholar]
  24. LoBuglio A. F., Court W. S., Vinocur L., Maglott G., Shaw G. M. Immune thrombocytopenic purpura. Use of a 125I-labeled antihuman IgG monoclonal antibody to quantify platelet-bound IgG. N Engl J Med. 1983 Aug 25;309(8):459–463. doi: 10.1056/NEJM198308253090804. [DOI] [PubMed] [Google Scholar]
  25. Luiken G. A., McMillan R., Lightsey A. L., Gordon P., Zevely S., Schulman I., Gribble T. J., Longmire R. L. Platelet-associated IgG in immune thrombocytopenic purpura. Blood. 1977 Aug;50(2):317–325. [PubMed] [Google Scholar]
  26. McGrath K. M., Stuart J. J., Richards F., 2nd Correlation between serum IgG, platelet membrane IgG, and platelet function in hypergammaglobulinaemic states. Br J Haematol. 1979 Aug;42(4):585–591. doi: 10.1111/j.1365-2141.1979.tb01171.x. [DOI] [PubMed] [Google Scholar]
  27. McMillan R. Chronic idiopathic thrombocytopenic purpura. N Engl J Med. 1981 May 7;304(19):1135–1147. doi: 10.1056/NEJM198105073041904. [DOI] [PubMed] [Google Scholar]
  28. McMillan R., Longmire R. L., Yelenosky R., Lang J. E., Heath V., Craddock C. G. Immunoglobulin synthesis by human lymphoid tissues: normal bone marrow as a major site of IgG production. J Immunol. 1972 Dec;109(6):1386–1394. [PubMed] [Google Scholar]
  29. McMillan R., Smith R. S., Longmire R. L., Yelenosky R., Reid R. T., Craddock C. G. Immunoglobulins associated with human platelets. Blood. 1971 Mar;37(3):316–322. [PubMed] [Google Scholar]
  30. Mueller-Eckhardt C., Mueller-Eckhardt G., Kayser W., Voss R. M., Wegner J., Küenzlen E. Platelet associated IgG, platelet survival, and platelet sequestration in thrombocytopenic states. Br J Haematol. 1982 Sep;52(1):49–58. doi: 10.1111/j.1365-2141.1982.tb03860.x. [DOI] [PubMed] [Google Scholar]
  31. Mustard J. F., Perry D. W., Ardlie N. G., Packham M. A. Preparation of suspensions of washed platelets from humans. Br J Haematol. 1972 Feb;22(2):193–204. doi: 10.1111/j.1365-2141.1972.tb08800.x. [DOI] [PubMed] [Google Scholar]
  32. Mustard J. F., Perry D. W., Kinlough-Rathbone R. L., Packham M. A. Factors responsible for ADP-induced release reaction of human platelets. Am J Physiol. 1975 Jun;228(6):1757–1765. doi: 10.1152/ajplegacy.1975.228.6.1757. [DOI] [PubMed] [Google Scholar]
  33. Nachman R., Levine R., Jaffe E. A. Synthesis of factor VIII antigen by cultured guinea pig megakaryocytes. J Clin Invest. 1977 Oct;60(4):914–921. doi: 10.1172/JCI108846. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Natvig J. B., Kunkel H. G. Human immunoglobulins: classes, subclasses, genetic variants, and idiotypes. Adv Immunol. 1973;16:1–59. doi: 10.1016/s0065-2776(08)60295-3. [DOI] [PubMed] [Google Scholar]
  35. Pfueller S. L., Cosgrove L., Firkin B. G., Tew D. Relationship of raised platelet IgG in thrombocytopenia to total platelet protein content. Br J Haematol. 1981 Oct;49(2):293–302. doi: 10.1111/j.1365-2141.1981.tb07226.x. [DOI] [PubMed] [Google Scholar]
  36. Rosse W. F., Adams J. P., Yount W. J. Subclasses of IgG antibodies in immune thrombocytopenic purpura (ITPO). Br J Haematol. 1980 Sep;46(1):109–114. doi: 10.1111/j.1365-2141.1980.tb05940.x. [DOI] [PubMed] [Google Scholar]
  37. Ryo R., Nakeff A., Huang S. S., Ginsberg M., Deuel T. F. New synthesis of a platelet-specific protein: platelet factor 4 synthesis in a megakaryocyte-enriched rabbit bone marrow culture system. J Cell Biol. 1983 Feb;96(2):515–520. doi: 10.1083/jcb.96.2.515. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. SALMON J. Etude immunoélectrophorétique des antigènes plaquettaires humains. Schweiz Med Wochenschr. 1958 Oct 18;88(42):1047–1049. [PubMed] [Google Scholar]
  39. STERLING K. The turnover rate of serum albumin in man as measured by I131-tagged albumin. J Clin Invest. 1951 Nov;30(11):1228–1237. doi: 10.1172/JCI102542. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Sander H. J., Slot J. W., Bouma B. N., Bolhuis P. A., Pepper D. S., Sixma J. J. Immunocytochemical localization of fibrinogen, platelet factor 4, and beta thromboglobulin in thin frozen sections of human blood platelets. J Clin Invest. 1983 Oct;72(4):1277–1287. doi: 10.1172/JCI111084. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Shaw G. M., Axelson J., Maglott J. G., LoBuglio A. F. Quantification of platelet-bound IgG by 125I-Staphylococcal protein A in immune thrombocytopenic purpura and other thrombocytopenic disorders. Blood. 1984 Jan;63(1):154–161. [PubMed] [Google Scholar]
  42. Shuman M. A., Levine S. P. Relationship between secretion of platelet Factor 4 and thrombin generation during in vitro blood clotting. J Clin Invest. 1980 Feb;65(2):307–313. doi: 10.1172/JCI109673. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Sixma J. J., van den Berg A., Schiphorst M., Geuze H. J., McDonagh J. Immunocytochemical localization of albumin and factor XIII in thin cryo sections of human blood platelets. Thromb Haemost. 1984 Jul 29;51(3):388–391. [PubMed] [Google Scholar]
  44. Stenberg P. E., Shuman M. A., Levine S. P., Bainton D. F. Optimal techniques for the immunocytochemical demonstration of beta-thromboglobulin, platelet factor 4, and fibrinogen in the alpha granules of unstimulated platelets. Histochem J. 1984 Sep;16(9):983–1001. doi: 10.1007/BF01003853. [DOI] [PubMed] [Google Scholar]
  45. Stenberg P. E., Shuman M. A., Levine S. P., Bainton D. F. Redistribution of alpha-granules and their contents in thrombin-stimulated platelets. J Cell Biol. 1984 Feb;98(2):748–760. doi: 10.1083/jcb.98.2.748. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Tokuyasu K. T. Immunochemistry on ultrathin frozen sections. Histochem J. 1980 Jul;12(4):381–403. doi: 10.1007/BF01011956. [DOI] [PubMed] [Google Scholar]
  47. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Waldmann T. A., Strober W. Metabolism of immunoglobulins. Prog Allergy. 1969;13:1–110. doi: 10.1159/000385919. [DOI] [PubMed] [Google Scholar]
  49. Weiss H. J., Witte L. D., Kaplan K. L., Lages B. A., Chernoff A., Nossel H. L., Goodman D. S., Baumgartner H. R. Heterogeneity in storage pool deficiency: studies on granule-bound substances in 18 patients including variants deficient in alpha-granules, platelet factor 4, beta-thromboglobulin, and platelet-derived growth factor. Blood. 1979 Dec;54(6):1296–1319. [PubMed] [Google Scholar]

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