Abstract
Thrombospondin (TSP) is a multifunctional platelet glycoprotein synthesized by a variety of cells in culture including monocytes and macrophages. We now report that 125I-TSP binds specifically, saturably, and reversibly to mouse peritoneal macrophages and to cells of the monocyte-like human cell line U937 with dissociation constants of 6.7-14.5 X 10(-8) M and 3-4 X 10(5) binding sites per cell. TSP mediates an adhesive interaction between thrombin-stimulated platelets and both U937 cells and human blood monocytes. Using a sensitive rosetting assay, we found that monocytes were not rosetted by resting platelets whereas greater than 90% were rosetted by thrombin-stimulated platelets. Monoclonal and polyclonal anti-TSP antibodies markedly inhibited rosetting as did TSP itself. Neither control antibodies nor heparin, fibronectin, fibrinogen, nor the fibronectin adhesion tetrapeptide Arg-Gly-Asp-Ser inhibited rosetting. TSP may thus serve as a molecular bridge linking activated platelets with monocytes at sites of early vascular injury. Such interaction may be of critical importance in the regulation of thrombosis and the initiation of atherosclerosis.
Full text
PDF![867](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ebbb/424223/45b99e2c6538/jcinvest00114-0207.png)
![868](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ebbb/424223/8052776ba4b1/jcinvest00114-0208.png)
![869](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ebbb/424223/2d1f5c826585/jcinvest00114-0209.png)
![870](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ebbb/424223/c5ee2baafa00/jcinvest00114-0210.png)
![871](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ebbb/424223/2b1fea60c120/jcinvest00114-0211.png)
![872](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ebbb/424223/d9387c87a5a8/jcinvest00114-0212.png)
![873](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ebbb/424223/c79b8dbffec9/jcinvest00114-0213.png)
![874](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ebbb/424223/a4838e704afe/jcinvest00114-0214.png)
Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Asch A. S., Leung L. L., Shapiro J., Nachman R. L. Human brain glial cells synthesize thrombospondin. Proc Natl Acad Sci U S A. 1986 May;83(9):2904–2908. doi: 10.1073/pnas.83.9.2904. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bennett J. S., Vilaire G. Exposure of platelet fibrinogen receptors by ADP and epinephrine. J Clin Invest. 1979 Nov;64(5):1393–1401. doi: 10.1172/JCI109597. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bohnsack J. F., Kleinman H. K., Takahashi T., O'Shea J. J., Brown E. J. Connective tissue proteins and phagocytic cell function. Laminin enhances complement and Fc-mediated phagocytosis by cultured human macrophages. J Exp Med. 1985 May 1;161(5):912–923. doi: 10.1084/jem.161.5.912. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Castellot J. J., Jr, Favreau L. V., Karnovsky M. J., Rosenberg R. D. Inhibition of vascular smooth muscle cell growth by endothelial cell-derived heparin. Possible role of a platelet endoglycosidase. J Biol Chem. 1982 Oct 10;257(19):11256–11260. [PubMed] [Google Scholar]
- DiCorleto P. E., de la Motte C. A. Characterization of the adhesion of the human monocytic cell line U937 to cultured endothelial cells. J Clin Invest. 1985 Apr;75(4):1153–1161. doi: 10.1172/JCI111810. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dixit V. M., Haverstick D. M., O'Rourke K. M., Hennessy S. W., Grant G. A., Santoro S. A., Frazier W. A. A monoclonal antibody against human thrombospondin inhibits platelet aggregation. Proc Natl Acad Sci U S A. 1985 May;82(10):3472–3476. doi: 10.1073/pnas.82.10.3472. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Faggiotto A., Ross R., Harker L. Studies of hypercholesterolemia in the nonhuman primate. I. Changes that lead to fatty streak formation. Arteriosclerosis. 1984 Jul-Aug;4(4):323–340. doi: 10.1161/01.atv.4.4.323. [DOI] [PubMed] [Google Scholar]
- Galvin N. J., Dixit V. M., O'Rourke K. M., Santoro S. A., Grant G. A., Frazier W. A. Mapping of epitopes for monoclonal antibodies against human platelet thrombospondin with electron microscopy and high sensitivity amino acid sequencing. J Cell Biol. 1985 Oct;101(4):1434–1441. doi: 10.1083/jcb.101.4.1434. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gerrity R. G. The role of the monocyte in atherogenesis: II. Migration of foam cells from atherosclerotic lesions. Am J Pathol. 1981 May;103(2):191–200. [PMC free article] [PubMed] [Google Scholar]
- Ginsberg M. H., Forsyth J., Lightsey A., Chediak J., Plow E. F. Reduced surface expression and binding of fibronectin by thrombin-stimulated thrombasthenic platelets. J Clin Invest. 1983 Mar;71(3):619–624. doi: 10.1172/JCI110808. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gospodarowicz D., Ill C. Extracellular matrix and control of proliferation of vascular endothelial cells. J Clin Invest. 1980 Jun;65(6):1351–1364. doi: 10.1172/JCI109799. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hagen I. Effects of thrombin on washed, human platelets: changes in the subcellular fractions. Biochim Biophys Acta. 1975 Jun 12;392(2):242–254. doi: 10.1016/0304-4165(75)90006-9. [DOI] [PubMed] [Google Scholar]
- Jaffe E. A., Leung L. L., Nachman R. L., Levin R. I., Mosher D. F. Thrombospondin is the endogenous lectin of human platelets. Nature. 1982 Jan 21;295(5846):246–248. doi: 10.1038/295246a0. [DOI] [PubMed] [Google Scholar]
- Jaffe E. A., Ruggiero J. T., Falcone D. J. Monocytes and macrophages synthesize and secrete thrombospondin. Blood. 1985 Jan;65(1):79–84. [PubMed] [Google Scholar]
- Jaffe E. A., Ruggiero J. T., Leung L. K., Doyle M. J., McKeown-Longo P. J., Mosher D. F. Cultured human fibroblasts synthesize and secrete thrombospondin and incorporate it into extracellular matrix. Proc Natl Acad Sci U S A. 1983 Feb;80(4):998–1002. doi: 10.1073/pnas.80.4.998. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jungi T. W., Spycher M. O., Nydegger U. E., Barandun S. Platelet-leukocyte interaction: selective binding of thrombin-stimulated platelets to human monocytes, polymorphonuclear leukocytes, and related cell lines. Blood. 1986 Mar;67(3):629–636. [PubMed] [Google Scholar]
- Kjeldsberg C. R., Swanson J. Platelet satellitism. Blood. 1974 Jun;43(6):831–836. [PubMed] [Google Scholar]
- Köhler G., Milstein C. Continuous cultures of fused cells secreting antibody of predefined specificity. Nature. 1975 Aug 7;256(5517):495–497. doi: 10.1038/256495a0. [DOI] [PubMed] [Google Scholar]
- Lahav J., Schwartz M. A., Hynes R. O. Analysis of platelet adhesion with a radioactive chemical crosslinking reagent: interaction of thrombospondin with fibronectin and collagen. Cell. 1982 Nov;31(1):253–262. doi: 10.1016/0092-8674(82)90425-1. [DOI] [PubMed] [Google Scholar]
- Lawler J. W., Slayter H. S., Coligan J. E. Isolation and characterization of a high molecular weight glycoprotein from human blood platelets. J Biol Chem. 1978 Dec 10;253(23):8609–8616. [PubMed] [Google Scholar]
- Leung L. L., Nachman R. L. Complex formation of platelet thrombospondin with fibrinogen. J Clin Invest. 1982 Sep;70(3):542–549. doi: 10.1172/JCI110646. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Leung L. L., Nachman R. L., Harpel P. C. Complex formation of platelet thrombospondin with histidine-rich glycoprotein. J Clin Invest. 1984 Jan;73(1):5–12. doi: 10.1172/JCI111206. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Leung L. L. Role of thrombospondin in platelet aggregation. J Clin Invest. 1984 Nov;74(5):1764–1772. doi: 10.1172/JCI111595. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Majack R. A., Cook S. C., Bornstein P. Platelet-derived growth factor and heparin-like glycosaminoglycans regulate thrombospondin synthesis and deposition in the matrix by smooth muscle cells. J Cell Biol. 1985 Sep;101(3):1059–1070. doi: 10.1083/jcb.101.3.1059. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mosher D. F., Doyle M. J., Jaffe E. A. Synthesis and secretion of thrombospondin by cultured human endothelial cells. J Cell Biol. 1982 May;93(2):343–348. doi: 10.1083/jcb.93.2.343. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mumby S. M., Abbott-Brown D., Raugi G. J., Bornstein P. Regulation of thrombospondin secretion by cells in culture. J Cell Physiol. 1984 Sep;120(3):280–288. doi: 10.1002/jcp.1041200304. [DOI] [PubMed] [Google Scholar]
- Mumby S. M., Raugi G. J., Bornstein P. Interactions of thrombospondin with extracellular matrix proteins: selective binding to type V collagen. J Cell Biol. 1984 Feb;98(2):646–652. doi: 10.1083/jcb.98.2.646. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Munson P. J., Rodbard D. Ligand: a versatile computerized approach for characterization of ligand-binding systems. Anal Biochem. 1980 Sep 1;107(1):220–239. doi: 10.1016/0003-2697(80)90515-1. [DOI] [PubMed] [Google Scholar]
- PORTER R. R. The hydrolysis of rabbit y-globulin and antibodies with crystalline papain. Biochem J. 1959 Sep;73:119–126. doi: 10.1042/bj0730119. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Perussia B., Jankiewicz J., Trinchieri G. Binding of platelets to human monocytes: a source of artifacts in the study of the specificity of antileukocyte antibodies. J Immunol Methods. 1982;50(3):269–276. doi: 10.1016/0022-1759(82)90164-8. [DOI] [PubMed] [Google Scholar]
- Pierschbacher M. D., Ruoslahti E. Cell attachment activity of fibronectin can be duplicated by small synthetic fragments of the molecule. Nature. 1984 May 3;309(5963):30–33. doi: 10.1038/309030a0. [DOI] [PubMed] [Google Scholar]
- Plow E. F., Srouji A. H., Meyer D., Marguerie G., Ginsberg M. H. Evidence that three adhesive proteins interact with a common recognition site on activated platelets. J Biol Chem. 1984 May 10;259(9):5388–5391. [PubMed] [Google Scholar]
- Ralph P., Williams N., Moore M. A., Litcofsky P. B. Induction of antibody-dependent and nonspecific tumor killing in human monocytic leukemia cells by nonlymphocyte factors and phorbol ester. Cell Immunol. 1982 Aug;71(2):215–223. doi: 10.1016/0008-8749(82)90257-x. [DOI] [PubMed] [Google Scholar]
- Raugi G. J., Mumby S. M., Abbott-Brown D., Bornstein P. Thrombospondin: synthesis and secretion by cells in culture. J Cell Biol. 1982 Oct;95(1):351–354. doi: 10.1083/jcb.95.1.351. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Robert A., Grunberger G., Carpenter J. L., Dayer J. M., Orci L., Gorden P. The insulin receptor of a human monocyte-like cell line: characterization and function. Endocrinology. 1984 Jan;114(1):247–253. doi: 10.1210/endo-114-1-247. [DOI] [PubMed] [Google Scholar]
- Roberts D. D., Haverstick D. M., Dixit V. M., Frazier W. A., Santoro S. A., Ginsburg V. The platelet glycoprotein thrombospondin binds specifically to sulfated glycolipids. J Biol Chem. 1985 Aug 5;260(16):9405–9411. [PubMed] [Google Scholar]
- Roberts D. D., Sherwood J. A., Spitalnik S. L., Panton L. J., Howard R. J., Dixit V. M., Frazier W. A., Miller L. H., Ginsburg V. Thrombospondin binds falciparum malaria parasitized erythrocytes and may mediate cytoadherence. Nature. 1985 Nov 7;318(6041):64–66. doi: 10.1038/318064a0. [DOI] [PubMed] [Google Scholar]
- Saglio S. D., Slayter H. S. Use of a radioimmunoassay to quantify thrombospondin. Blood. 1982 Jan;59(1):162–166. [PubMed] [Google Scholar]
- Silverstein R. L., Leung L. L., Harpel P. C., Nachman R. L. Complex formation of platelet thrombospondin with plasminogen. Modulation of activation by tissue activator. J Clin Invest. 1984 Nov;74(5):1625–1633. doi: 10.1172/JCI111578. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Silverstein R. L., Leung L. L., Harpel P. C., Nachman R. L. Platelet thrombospondin forms a trimolecular complex with plasminogen and histidine-rich glycoprotein. J Clin Invest. 1985 Jun;75(6):2065–2073. doi: 10.1172/JCI111926. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Silverstein R. L., Leung L. L., Nachman R. L. Thrombospondin: a versatile multifunctional glycoprotein. Arteriosclerosis. 1986 May-Jun;6(3):245–253. doi: 10.1161/01.atv.6.3.245. [DOI] [PubMed] [Google Scholar]
- Silverstein R. L., Nachman R. L., Leung L. L., Harpel P. C. Activation of immobilized plasminogen by tissue activator. Multimolecular complex formation. J Biol Chem. 1985 Aug 25;260(18):10346–10352. [PubMed] [Google Scholar]
- Sundström C., Nilsson K. Establishment and characterization of a human histiocytic lymphoma cell line (U-937). Int J Cancer. 1976 May 15;17(5):565–577. doi: 10.1002/ijc.2910170504. [DOI] [PubMed] [Google Scholar]
- 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]
- Tracy P. B., Eide L. L., Mann K. G. Human prothrombinase complex assembly and function on isolated peripheral blood cell populations. J Biol Chem. 1985 Feb 25;260(4):2119–2124. [PubMed] [Google Scholar]
- Unkeless J. C., Gordon S., Reich E. Secretion of plasminogen activator by stimulated macrophages. J Exp Med. 1974 Apr 1;139(4):834–850. doi: 10.1084/jem.139.4.834. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wight T. N., Raugi G. J., Mumby S. M., Bornstein P. Light microscopic immunolocation of thrombospondin in human tissues. J Histochem Cytochem. 1985 Apr;33(4):295–302. doi: 10.1177/33.4.3884704. [DOI] [PubMed] [Google Scholar]
- Wolff R., Plow E. F., Ginsberg M. H. Interaction of thrombospondin with resting and stimulated human platelets. J Biol Chem. 1986 May 25;261(15):6840–6846. [PubMed] [Google Scholar]
- Wright S. D., Silverstein S. C. Tumor-promoting phorbol esters stimulate C3b and C3b' receptor-mediated phagocytosis in cultured human monocytes. J Exp Med. 1982 Oct 1;156(4):1149–1164. doi: 10.1084/jem.156.4.1149. [DOI] [PMC free article] [PubMed] [Google Scholar]