Aggregation of chymotrypsin-treated thrombasthenic platelets is mediated by fibrinogen binding to glycoproteins IIb and IIIa

Stefan Niewiarowski, Elizabeth Kornecki, Diane Hershock, George P. Tuszynski, Joel S. Bennett, Claudine Soria, Jeanette Soria, Fred Dunn, Dominique Pidard, Nelly Kieffer, Alan T. Nurden

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Abstract

Previous experiments demonstrated that chymotrypsin, but not adenosine diphosphate (ADP), exposed fibrinogen binding sites on platelets from patients with Glanzmann's thrombasthenia. Three of these patients have been reexamined, and previous observations were confirmed. The quantity of iodine 125-labeled glycoprotein IIb (GPIIb) and glycoprotein IIIa [GPIIIa) on the platelets of these patients was considerably less than normal but was detectable by immunoprecipitation, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and autoradiography. The amount of residual GPIIb and GPIIIa as measured by binding studies with radiolabeled monoclonal antibodies was between 3% and 12% of the normal value. Platelet suspensions from these patients did not aggregate with fibrinogen and did not bind 125I-fibrinogen on stimulation with ADP. However, incubation of these platelets with chymotrypsin or pronase resulted in fibrinogen binding and platelet aggregation. Monoclonal antibodies specific for the GPIIb-GPIIIa complex blocked both the fibrinogen binding and the aggregation of enzyme-treated platelets. The treatment of washed platelets of a fourth thrombasthenlc patient with ADP or with chymotrypsin failed to result in fibrinogen binding and aggregation. However, the level of GPIIb and GPIIIa on these platelets as measured by a Western blot technique and by monoclonal antibody binding amounted to <0.35% to 0.5% of normal values. In conclusion, fibrinogen binding sites exposed on thrombasthenic platelets by chymotrypsin are derived from GPIIb-GPIIIa molecules. Aggregation of chymotrypsin-treated thrombasthenic platelets by fibrinogen appears to represent a sensitive test for detection of functionally active GPIIb-GPIIIa complex on the platelet surface.

Original languageEnglish (US)
Pages (from-to)651-660
Number of pages10
JournalThe Journal of Laboratory and Clinical Medicine
Volume106
Issue number6
StatePublished - Dec 1985

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Platelet Glycoprotein GPIIb-IIIa Complex
Chymotrypsin
Platelets
Fibrinogen
Blood Platelets
Agglomeration
Glycoproteins
Adenosine Diphosphate
Monoclonal Antibodies
Reference Values
Binding Sites
Thrombasthenia
Pronase
Autoradiography
Platelet Aggregation
Immunoprecipitation
Sodium Dodecyl Sulfate
Iodine
Electrophoresis
Polyacrylamide Gel Electrophoresis

All Science Journal Classification (ASJC) codes

  • Pathology and Forensic Medicine

Cite this

Niewiarowski, S., Kornecki, E., Hershock, D., Tuszynski, G. P., Bennett, J. S., Soria, C., ... Nurden, A. T. (1985). Aggregation of chymotrypsin-treated thrombasthenic platelets is mediated by fibrinogen binding to glycoproteins IIb and IIIa. The Journal of Laboratory and Clinical Medicine, 106(6), 651-660.
Niewiarowski, Stefan ; Kornecki, Elizabeth ; Hershock, Diane ; Tuszynski, George P. ; Bennett, Joel S. ; Soria, Claudine ; Soria, Jeanette ; Dunn, Fred ; Pidard, Dominique ; Kieffer, Nelly ; Nurden, Alan T. / Aggregation of chymotrypsin-treated thrombasthenic platelets is mediated by fibrinogen binding to glycoproteins IIb and IIIa. In: The Journal of Laboratory and Clinical Medicine. 1985 ; Vol. 106, No. 6. pp. 651-660.
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abstract = "Previous experiments demonstrated that chymotrypsin, but not adenosine diphosphate (ADP), exposed fibrinogen binding sites on platelets from patients with Glanzmann's thrombasthenia. Three of these patients have been reexamined, and previous observations were confirmed. The quantity of iodine 125-labeled glycoprotein IIb (GPIIb) and glycoprotein IIIa [GPIIIa) on the platelets of these patients was considerably less than normal but was detectable by immunoprecipitation, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and autoradiography. The amount of residual GPIIb and GPIIIa as measured by binding studies with radiolabeled monoclonal antibodies was between 3{\%} and 12{\%} of the normal value. Platelet suspensions from these patients did not aggregate with fibrinogen and did not bind 125I-fibrinogen on stimulation with ADP. However, incubation of these platelets with chymotrypsin or pronase resulted in fibrinogen binding and platelet aggregation. Monoclonal antibodies specific for the GPIIb-GPIIIa complex blocked both the fibrinogen binding and the aggregation of enzyme-treated platelets. The treatment of washed platelets of a fourth thrombasthenlc patient with ADP or with chymotrypsin failed to result in fibrinogen binding and aggregation. However, the level of GPIIb and GPIIIa on these platelets as measured by a Western blot technique and by monoclonal antibody binding amounted to <0.35{\%} to 0.5{\%} of normal values. In conclusion, fibrinogen binding sites exposed on thrombasthenic platelets by chymotrypsin are derived from GPIIb-GPIIIa molecules. Aggregation of chymotrypsin-treated thrombasthenic platelets by fibrinogen appears to represent a sensitive test for detection of functionally active GPIIb-GPIIIa complex on the platelet surface.",
author = "Stefan Niewiarowski and Elizabeth Kornecki and Diane Hershock and Tuszynski, {George P.} and Bennett, {Joel S.} and Claudine Soria and Jeanette Soria and Fred Dunn and Dominique Pidard and Nelly Kieffer and Nurden, {Alan T.}",
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Niewiarowski, S, Kornecki, E, Hershock, D, Tuszynski, GP, Bennett, JS, Soria, C, Soria, J, Dunn, F, Pidard, D, Kieffer, N & Nurden, AT 1985, 'Aggregation of chymotrypsin-treated thrombasthenic platelets is mediated by fibrinogen binding to glycoproteins IIb and IIIa', The Journal of Laboratory and Clinical Medicine, vol. 106, no. 6, pp. 651-660.

Aggregation of chymotrypsin-treated thrombasthenic platelets is mediated by fibrinogen binding to glycoproteins IIb and IIIa. / Niewiarowski, Stefan; Kornecki, Elizabeth; Hershock, Diane; Tuszynski, George P.; Bennett, Joel S.; Soria, Claudine; Soria, Jeanette; Dunn, Fred; Pidard, Dominique; Kieffer, Nelly; Nurden, Alan T.

In: The Journal of Laboratory and Clinical Medicine, Vol. 106, No. 6, 12.1985, p. 651-660.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Aggregation of chymotrypsin-treated thrombasthenic platelets is mediated by fibrinogen binding to glycoproteins IIb and IIIa

AU - Niewiarowski, Stefan

AU - Kornecki, Elizabeth

AU - Hershock, Diane

AU - Tuszynski, George P.

AU - Bennett, Joel S.

AU - Soria, Claudine

AU - Soria, Jeanette

AU - Dunn, Fred

AU - Pidard, Dominique

AU - Kieffer, Nelly

AU - Nurden, Alan T.

PY - 1985/12

Y1 - 1985/12

N2 - Previous experiments demonstrated that chymotrypsin, but not adenosine diphosphate (ADP), exposed fibrinogen binding sites on platelets from patients with Glanzmann's thrombasthenia. Three of these patients have been reexamined, and previous observations were confirmed. The quantity of iodine 125-labeled glycoprotein IIb (GPIIb) and glycoprotein IIIa [GPIIIa) on the platelets of these patients was considerably less than normal but was detectable by immunoprecipitation, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and autoradiography. The amount of residual GPIIb and GPIIIa as measured by binding studies with radiolabeled monoclonal antibodies was between 3% and 12% of the normal value. Platelet suspensions from these patients did not aggregate with fibrinogen and did not bind 125I-fibrinogen on stimulation with ADP. However, incubation of these platelets with chymotrypsin or pronase resulted in fibrinogen binding and platelet aggregation. Monoclonal antibodies specific for the GPIIb-GPIIIa complex blocked both the fibrinogen binding and the aggregation of enzyme-treated platelets. The treatment of washed platelets of a fourth thrombasthenlc patient with ADP or with chymotrypsin failed to result in fibrinogen binding and aggregation. However, the level of GPIIb and GPIIIa on these platelets as measured by a Western blot technique and by monoclonal antibody binding amounted to <0.35% to 0.5% of normal values. In conclusion, fibrinogen binding sites exposed on thrombasthenic platelets by chymotrypsin are derived from GPIIb-GPIIIa molecules. Aggregation of chymotrypsin-treated thrombasthenic platelets by fibrinogen appears to represent a sensitive test for detection of functionally active GPIIb-GPIIIa complex on the platelet surface.

AB - Previous experiments demonstrated that chymotrypsin, but not adenosine diphosphate (ADP), exposed fibrinogen binding sites on platelets from patients with Glanzmann's thrombasthenia. Three of these patients have been reexamined, and previous observations were confirmed. The quantity of iodine 125-labeled glycoprotein IIb (GPIIb) and glycoprotein IIIa [GPIIIa) on the platelets of these patients was considerably less than normal but was detectable by immunoprecipitation, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and autoradiography. The amount of residual GPIIb and GPIIIa as measured by binding studies with radiolabeled monoclonal antibodies was between 3% and 12% of the normal value. Platelet suspensions from these patients did not aggregate with fibrinogen and did not bind 125I-fibrinogen on stimulation with ADP. However, incubation of these platelets with chymotrypsin or pronase resulted in fibrinogen binding and platelet aggregation. Monoclonal antibodies specific for the GPIIb-GPIIIa complex blocked both the fibrinogen binding and the aggregation of enzyme-treated platelets. The treatment of washed platelets of a fourth thrombasthenlc patient with ADP or with chymotrypsin failed to result in fibrinogen binding and aggregation. However, the level of GPIIb and GPIIIa on these platelets as measured by a Western blot technique and by monoclonal antibody binding amounted to <0.35% to 0.5% of normal values. In conclusion, fibrinogen binding sites exposed on thrombasthenic platelets by chymotrypsin are derived from GPIIb-GPIIIa molecules. Aggregation of chymotrypsin-treated thrombasthenic platelets by fibrinogen appears to represent a sensitive test for detection of functionally active GPIIb-GPIIIa complex on the platelet surface.

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