Assessment of Lumiaggregometry for Research and Clinical Laboratories

Summary

Platelet aggregometry is often used to help diagnose storage pool disease (SPD-reduced amounts of granule nucleotides) and release defects (abnormal release of granule nucleotides). The general assumption that normal aggregation patterns are sufficient to rule out the diagnosis of one of these disorders has been invalidated by the recent publication of two papers describing patients with clinical bleeding, prolonged bleeding times and normal aggregation patterns in spite of defective release. The lumiaggregometer provides a tool for measuring platelet release and aggregation simultaneously. This paper presents a standardized, reproducible method for the use of the lumiaggregometer based on a “standard curve”. Data obtained during the development of the procedure are presented including normal ranges of release at different concentrations of agonists, release measured in intrinsic disorders as well as in patients on aspirin, and values for release relative to varying platelet counts. A monoclonal antibody (anti-p24/CD9; MAb7) which activates platelets similarly to thrombin and may be a useful reagent for distinguishing SPD and release defects is also introduced.

• References

• 1 Borchgrevink CF. A method for measuring platelet adhesiveness in vivo. Acta Med Scand 1960; 168: 157-164
• 2 Salzman EW. Measurement of platelet adhesiveness. A simple in vitro technique demonstrating an abnormality in von Willebrand’s disease. J Lab Clin Med 1963; 62: 724-735
• 3 Helium AJ. Platelet adhesiveness. Ser Haematol 1968; 1 (02) 99-145
• 4 Bowie EJW, Owen CA, Thompson JH, Didisheim P. Platelet adhesiveness in von Willebrand’s disease. Am J Clin Pathol 1969; 52: 69-77
• 5 Rossi EC, Green D. A study of platelet retention by glass bead columns (“platelet adhesiveness” in normal subjects). Br J Haematol 1972; 23: 47-57
• 6 Hardisty RM, Dormandy KM, Hutton RA. Thrombasthenia. Studies on three cases. Br J Haematol 1964; 10: 371-387
• 7 Nurden AT, Caen JP. An abnormal platelet glycoprotein pattern in three cases of Glanzmann’s thrombasthenia. Br J Haematol 1974; 23: 253-260
• 8 Phillips DR, Jenkins CSP, Luscher EF, Larrieu MJ. Molecular differences of exposed surface proteins on thrombasthenic platelet plasma membranes. Nature 1975; 257: 599-600
• 9 Holmsen H, Weiss HJ. Further evidence for a deficient storage pool of adenine nucleotides in platelets from some patients with thrombocytopenia - “Storage pool disease”. Blood 1972; 39: 197-209
• 10 Weiss HJ, Rogers J. Thrombocytopathia due to abnormalities in platelet release reaction - studies on six unrelated patients. Blood 1972; 39: 187-196
• 11 Weiss HJ. Abnormalities in platelet function due to defects in the release reaction. Ann NY Acad Sci 1972; 201: 161-173
• 12 Sahud MA, Aggeler PM. Platelet dysfunction: Differentiation of a newly recognized primary type from that produced by aspirin. N Engl J Med 1969; 280: 453-459
• 13 Feinman RD, Detwiler TC, Ingerman-Wojenski C. The lumi-aggregometer as a research and clinical tool. In: The Platelets: Physiology and Pharmacology. Longenecker GL. (ed) Academic Press; San Diego, CA: 1985. pp 429-440
• 14 Holmsen H, Storm E, Day HJ. Determination of ATP and ADP in blood platelets: a modification of the firefly luciferase assay for plasma. Anal Biochem 1972; 46: 489-501
• 15 Israelis SJ, McNicol A, Robertson C, Gerrard JM. Platelet storage pool deficiency: diagnosis in patients with prolonged bleeding times and normal platelet aggregation. Br J Haematol 1990; 75: 118-121
• 16 Nieuwenhuis HK, Akkerman JWN, Sixma JJ. Patients with a prolonged bleeding time and normal aggregation tests may have storage pool deficiency: studies on one hundred six patients. Blood 1987; 70 (03) 620-623
• 17 Jennings LK, Fox CF, Kouns WC, McKay CP, Ballou LR, Schultz HE. The activation of human platelets mediated by antihuman platelet p24/CD9 monoclonal antibodies. J Biol Chem 1990; 265: 3815-3822
• 18 Feinmann RD, Kubowsky J, Charo I, Zabinski MP. The lumi-aggregometer: a new instrument for simultaneous measurement of secretion and aggregation by platelets. J Lab Clin Med 1977; 90 (01) 125-129
• 19 Lofvenberg E, Nilsson TK. Qualitative platelet defects in chronic myeloproliferative disorders: evidence for reduced ATP secretion. Eur J Haematol 1989; 43: 435-440
• 20 Panella TJ, Peters W, White JG, Hannun YA, Greenberg CS. Platelets acquire a secretion defect after high-dose chemotherapy. Cancer 1990; 65: 1711-1716
• 21 Balduini CL, Bertolino G, Noris P, Sinigaglia F, Bisio A, Torti M. Interrelation of platelet aggregation, release reaction and thromboxane A2 production. Biochem Biophys Res Commun 1988; 156 (02) 823-829
• 22 Horellou MH, Lecompte T, Lecrubier C, Fouque F, Chignard M, Conard J, Vargaftig BB, Dray F, Samama M. Familial and constitutional bleeding disorder due to platelet cyclo-oxygenase deficiency. Am J Hematol 1983; 14: 1-9
• 23 Balduini CL, Fabiola S, Damiani G, Bertoline G, Bisio A, Zocchi E. Effect of a monoclonal antibody against GP’s Ilb-IIIa on platelet aggregation and ATP secretion. Exp Hematol 1986; 14: 338-342
• 24 Hannun YA, Greenberg CS, Bell RM. Sphingosine Inhibition of agonist-dependent secretion and activation of human platelets implies that protein kinase C is a necessary and common event of the signal transduction pathways. J Biol Chem 1987; 262 (28) 13620-13626
• 25 Rink TJ, Sanchez A, Hallam TF. Diacylglycerol and phorbol ester stimulate secretion without raising cytoplasmic free calcium in human platelets. Nature 1983; 305: 317-319
• 26 Ingerman-Wojenski CM. Simultaneous measurement of platelet aggregation and the release reaction in platelet-rich plasma and in whole blood. J Med Tech 1984; 9: 697-701
• 27 Corriveau DM, Fritsma GA. (eds) Hemostasis in the clinical laboratory. JB Lippincott Company; Philadelphia, PA: 1988. p 254
• 28 Bousser MG, Eschwege E, Haguenau M. et al “AICLA” controlled trial of aspirin and dipyridamole in the secondary prevention of athero-thrombotic cerebral ischemia. Stroke 1983; 14 (01) 5-14
• 29 Lewis HD, Davis JW, Archibald DG. et al Protective effects of aspirin against acute myocardial infarction and death in men with unstable angina. N Engl J Med 1983; 309 (07) 396-403
• 30 Canner PL. Aspirin in coronary heart disease: comparison of six clinical trials. Isr J Med Sci 1983; 19: 413-423
• 31 Belanger C, Buring JE, Cook N. et al Preliminary report; findings from the aspirin component of the ongoing physicians’ health study. N Engl J Med 1988; 318 (04) 262-264