Effect of Anti-Thrombospondin Antibodies on the Hemagglutination Activities of the Endogenous Platelet Lectin and Thrombospondin

T Kent Gartner; Mary Jean Doyle; Deane F Mosher

doi:10.1055/s-0038-1661213

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 1984; 52(03): 354-357
DOI: 10.1055/s-0038-1661213

Original Article

Schattauer GmbH Stuttgart

Effect of Anti-Thrombospondin Antibodies on the Hemagglutination Activities of the Endogenous Platelet Lectin and Thrombospondin

T Kent Gartner

The Department of Biology, Memphis State University Memphis, TN, USA, and the Department of Medicine, University of Wisconsin-Madison, Madison, WI, USA

,

Mary Jean Doyle

The Department of Biology, Memphis State University Memphis, TN, USA, and the Department of Medicine, University of Wisconsin-Madison, Madison, WI, USA

,

Deane F Mosher

The Department of Biology, Memphis State University Memphis, TN, USA, and the Department of Medicine, University of Wisconsin-Madison, Madison, WI, USA

› Author Affiliations

Abstract

PDF Download

Summary

The proposal that thrombospondin is the endogenous platelet lectin was evaluated using antisera and monoclonal antibodies to thrombospondin. The platelet-bound hemagglutinin activity of human platelets stimulated with A23187 was inhibited by rabbit anti-thrombospondin sera and by a monoclonal anti-thrombospondin IgG. A second monoclonal IgG did not inhibit platelet- bound agglutinin activity. Preparations of purified platelet thrombospondin differed in their hemagglutination activities. The hemagglutination activity of an active preparation of thrombospondin was inhibited by the monoclonal antibody that inhibited platelet-bound lectin activity. The hemagglutination activity of an almost inactive preparation of thrombospondin was enhanced by the anti-thrombospondin monoclonal antibody that did not block platelet-bound lectin activity. The results demonstrate that expression of the platelet-bound form of the endogenous lectin is thrombospondin-dependent and suggest that thrombospondin must become part of a larger complex, either by binding to the platelet surface or by becoming aggregated in solution, before hemagglutination activity can be expressed.

Key words

Antibodies - Platelet lectin - Thrombospondin

PDF (555 kb)

References

References
1 Gartner TK, Williams DC, Minion FC, Phillips DR. Thrombin- induced platelet aggregation is mediated by a platelet plasma membrane-bound lectin. Science 1978; 200: 1281-1283
2 Gartner TK, Phillips DR, Williams DC. Expression of thrombin- enhanced platelet lectin activity is controlled by secretion. FEBS Lett 1980; 113: 196-200
3 Gartner TK, Gerrard JM, White JG, Williams DC. The endogenous lectin of human platelets is an a-granule component. Blood 1981; 58: 153-157
4 Gartner TK, Walz DA. TFP inhibits the expression of the endogenous platelet lectin and secretion of a-granules. Thromb Res 1983; 29: 63-74
5 Jaffe EA, Leung LL, Nachman RL, Levin RI, Mosher DF. Thrombospondin is the endogenous lectin of human platelets. Nature 1982; 295: 246-248
6 Lawler JW, Slayter HS, Coligan JE. Isolation and characterization of a high molecular weight glycoprotein from human blood platelets. J Biol Chem 1978; 253: 8609-8616
7 George JN. Studies on platelet plasma membranes. Quantitative analysis of platelet glycoproteins by (¹²⁵I) - diazotized diiodosulfanilic acid labeling and SDS - polyacrylamide gel electrophoresis J Lab Clin Med 1978; 92: 430-446
8 Hagan I. Effects of thrombin on washed, human platelets: changes in subcellular fractions. Biochim Biophys Acta 1975; 392: 242-254
9 George JN, Lyons RM, Morgan RK. Membrane changes associated with activation-exposure of actin on the platelet surface after throm-bin-induced secretion. J Clin Invest 1980; 66: 1-9
10 Phillips DR, Jennings LK, Prasanna HR. Ca²⁺ mediated association of glycoprotein G (thrombin sensitive protein, thrombospondin) with human platelets. J Biol Chem 1980; 255: 11629-11632
11 Gartner TK, Dockter ME. Secreted platelet thrombospondin binds monovalently to platelets and erythrocytes in the absence of free Ca2+. Thromb Res 1983; 33: 19-30
12 Bemdt MC, Booth WJ, Castaldi PA. Thrombospondin is not the endogenous lectin of human platelets. Thromb Haemostas 1983; 50: 126 (Abstr.)
13 Booth WJ, Furby FH, Berndt MC, Costaldi PA. Factor VIII/von Willebrand Factor has potent lectin activity. Biochem Biophys Res Commun 1984; 118: 495-501
14 Alexander RJ, Detwiler TC. Quantitative adsorption of platelet glycoprotein G (thrombin-sensitive protein, thrombospondin) to barium citrate. Biochem J 1984; 217: 67-71
15 Phillips DR, Agin PP. Platelet membrane defects in Glanzmann’s thrombasthenia. J Clin Invest 1977; 60: 535-545
16 Mosher DF, Doyle MJ, Jaffe EA. Synthesis and secretion of thrombospondin by cultured human endothelial cells. J Cell Biol 1982; 93: 343-348
17 Jaffe EA, Ruggiero JT, Leung LL K, Doyle MJ, McKeown-Longo PJ, Mosher DF. Cultured human fibroblasts synthesize and secrete thrombospondin and incorporate it into extracellular matrix. Proc Natl Acad Sci USA 1983; 80: 998-1002
18 Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein dye binding. Anal Biochem 1976; 72: 248-254
19 Saglio SD, Slayter HS. Use of a radioimmunoassay to quantify thrombospondin. Blood 1982; 59: 162-166
20 Nurden AT, Hasitz M, Rosa EP. Inhibition of thrombin-induced platelet aggregation by a rabbit antibody against human platelet thrombospondin. Thromb Haemostas 1983; 50: 132 (Abstr.)
21 Leung LL, Polley M, Nachman RL. The role of thrombospondin in platelet aggregation. Blood 1983; 62 (suppl) (Suppl. 01) 261 a