Mechanisms of hemolysis-associated platelet activation

C. C. Helms, M. Marvel, W. Zhao, M. Stahle, R. Vest, G. J. Kato, J. S. Lee, G. Christ, M. T. Gladwin, R. R. Hantgan, D. B. Kim-Shapiro

Research output: Contribution to journalArticle

Abstract

Summary: Background: Intravascular hemolysis occurs after blood transfusion, in hemolytic anemias, and in other conditions, and is associated with hypercoagulable states. Hemolysis has been shown to potently activate platelets in vitro and in vivo, and several mechanisms have been suggested to account for this, including: (i) direct activation by hemoglobin (Hb); (ii) increase in reactive oxygen species (ROS); (iii) scavenging of nitric oxide (NO) by released Hb; and (iv) release of intraerythrocytic ADP. Objective: To elucidate the mechanism of hemolysis-mediated platelet activation. Methods: We used flow cytometry to detect PAC-1 binding to activated platelets for in vitro experiments, and a Siemens' Advia 120 hematology system to assess platelet aggregation by using platelet counts from in vivo experiments in a rodent model. Results: We found that Hb did not directly activate platelets. However, ADP bound to Hb could cause platelet activation. Furthermore, platelet activation caused by shearing of red blood cells (RBCs) was reduced in the presence of apyrase, which metabolizes ADP to AMP. The use of ROS scavengers did not affect platelet activation. We also found that cell-free Hb enhanced platelet activation by abrogating the inhibitory effect of NO on platelet activation. In vivo infusions of ADP and purified (ADP-free) Hb, as well as hemolysate, resulted in platelet aggregation, as shown by decreased platelet counts. Conclusion: Two primary mechanisms account for RBC hemolysis-associated platelet activation: ADP release, which activates platelets; and cell-free Hb release, which enhances platelet activation by lowering NO bioavailability.

Original languageEnglish (US)
Pages (from-to)2148-2154
Number of pages7
JournalJournal of Thrombosis and Haemostasis
Volume11
Issue number12
DOIs
StatePublished - Dec 2013
Externally publishedYes

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Platelet Activation
Hemolysis
Adenosine Diphosphate
Hemoglobins
Blood Platelets
Platelet Count
Platelet Aggregation
Reactive Oxygen Species
Nitric Oxide
Erythrocytes
Apyrase
Hemolytic Anemia
Hematology
Adenosine Monophosphate
Blood Transfusion
Biological Availability
Rodentia
Flow Cytometry

Keywords

  • Hemoglobin
  • Hemolysis
  • Nitric oxide
  • Platelets
  • Red blood cells

ASJC Scopus subject areas

  • Hematology

Cite this

Helms, C. C., Marvel, M., Zhao, W., Stahle, M., Vest, R., Kato, G. J., ... Kim-Shapiro, D. B. (2013). Mechanisms of hemolysis-associated platelet activation. Journal of Thrombosis and Haemostasis, 11(12), 2148-2154. https://doi.org/10.1111/jth.12422

Mechanisms of hemolysis-associated platelet activation. / Helms, C. C.; Marvel, M.; Zhao, W.; Stahle, M.; Vest, R.; Kato, G. J.; Lee, J. S.; Christ, G.; Gladwin, M. T.; Hantgan, R. R.; Kim-Shapiro, D. B.

In: Journal of Thrombosis and Haemostasis, Vol. 11, No. 12, 12.2013, p. 2148-2154.

Research output: Contribution to journalArticle

Helms, CC, Marvel, M, Zhao, W, Stahle, M, Vest, R, Kato, GJ, Lee, JS, Christ, G, Gladwin, MT, Hantgan, RR & Kim-Shapiro, DB 2013, 'Mechanisms of hemolysis-associated platelet activation', Journal of Thrombosis and Haemostasis, vol. 11, no. 12, pp. 2148-2154. https://doi.org/10.1111/jth.12422
Helms CC, Marvel M, Zhao W, Stahle M, Vest R, Kato GJ et al. Mechanisms of hemolysis-associated platelet activation. Journal of Thrombosis and Haemostasis. 2013 Dec;11(12):2148-2154. https://doi.org/10.1111/jth.12422
Helms, C. C. ; Marvel, M. ; Zhao, W. ; Stahle, M. ; Vest, R. ; Kato, G. J. ; Lee, J. S. ; Christ, G. ; Gladwin, M. T. ; Hantgan, R. R. ; Kim-Shapiro, D. B. / Mechanisms of hemolysis-associated platelet activation. In: Journal of Thrombosis and Haemostasis. 2013 ; Vol. 11, No. 12. pp. 2148-2154.
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abstract = "Summary: Background: Intravascular hemolysis occurs after blood transfusion, in hemolytic anemias, and in other conditions, and is associated with hypercoagulable states. Hemolysis has been shown to potently activate platelets in vitro and in vivo, and several mechanisms have been suggested to account for this, including: (i) direct activation by hemoglobin (Hb); (ii) increase in reactive oxygen species (ROS); (iii) scavenging of nitric oxide (NO) by released Hb; and (iv) release of intraerythrocytic ADP. Objective: To elucidate the mechanism of hemolysis-mediated platelet activation. Methods: We used flow cytometry to detect PAC-1 binding to activated platelets for in vitro experiments, and a Siemens' Advia 120 hematology system to assess platelet aggregation by using platelet counts from in vivo experiments in a rodent model. Results: We found that Hb did not directly activate platelets. However, ADP bound to Hb could cause platelet activation. Furthermore, platelet activation caused by shearing of red blood cells (RBCs) was reduced in the presence of apyrase, which metabolizes ADP to AMP. The use of ROS scavengers did not affect platelet activation. We also found that cell-free Hb enhanced platelet activation by abrogating the inhibitory effect of NO on platelet activation. In vivo infusions of ADP and purified (ADP-free) Hb, as well as hemolysate, resulted in platelet aggregation, as shown by decreased platelet counts. Conclusion: Two primary mechanisms account for RBC hemolysis-associated platelet activation: ADP release, which activates platelets; and cell-free Hb release, which enhances platelet activation by lowering NO bioavailability.",
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AU - Helms, C. C.

AU - Marvel, M.

AU - Zhao, W.

AU - Stahle, M.

AU - Vest, R.

AU - Kato, G. J.

AU - Lee, J. S.

AU - Christ, G.

AU - Gladwin, M. T.

AU - Hantgan, R. R.

AU - Kim-Shapiro, D. B.

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N2 - Summary: Background: Intravascular hemolysis occurs after blood transfusion, in hemolytic anemias, and in other conditions, and is associated with hypercoagulable states. Hemolysis has been shown to potently activate platelets in vitro and in vivo, and several mechanisms have been suggested to account for this, including: (i) direct activation by hemoglobin (Hb); (ii) increase in reactive oxygen species (ROS); (iii) scavenging of nitric oxide (NO) by released Hb; and (iv) release of intraerythrocytic ADP. Objective: To elucidate the mechanism of hemolysis-mediated platelet activation. Methods: We used flow cytometry to detect PAC-1 binding to activated platelets for in vitro experiments, and a Siemens' Advia 120 hematology system to assess platelet aggregation by using platelet counts from in vivo experiments in a rodent model. Results: We found that Hb did not directly activate platelets. However, ADP bound to Hb could cause platelet activation. Furthermore, platelet activation caused by shearing of red blood cells (RBCs) was reduced in the presence of apyrase, which metabolizes ADP to AMP. The use of ROS scavengers did not affect platelet activation. We also found that cell-free Hb enhanced platelet activation by abrogating the inhibitory effect of NO on platelet activation. In vivo infusions of ADP and purified (ADP-free) Hb, as well as hemolysate, resulted in platelet aggregation, as shown by decreased platelet counts. Conclusion: Two primary mechanisms account for RBC hemolysis-associated platelet activation: ADP release, which activates platelets; and cell-free Hb release, which enhances platelet activation by lowering NO bioavailability.

AB - Summary: Background: Intravascular hemolysis occurs after blood transfusion, in hemolytic anemias, and in other conditions, and is associated with hypercoagulable states. Hemolysis has been shown to potently activate platelets in vitro and in vivo, and several mechanisms have been suggested to account for this, including: (i) direct activation by hemoglobin (Hb); (ii) increase in reactive oxygen species (ROS); (iii) scavenging of nitric oxide (NO) by released Hb; and (iv) release of intraerythrocytic ADP. Objective: To elucidate the mechanism of hemolysis-mediated platelet activation. Methods: We used flow cytometry to detect PAC-1 binding to activated platelets for in vitro experiments, and a Siemens' Advia 120 hematology system to assess platelet aggregation by using platelet counts from in vivo experiments in a rodent model. Results: We found that Hb did not directly activate platelets. However, ADP bound to Hb could cause platelet activation. Furthermore, platelet activation caused by shearing of red blood cells (RBCs) was reduced in the presence of apyrase, which metabolizes ADP to AMP. The use of ROS scavengers did not affect platelet activation. We also found that cell-free Hb enhanced platelet activation by abrogating the inhibitory effect of NO on platelet activation. In vivo infusions of ADP and purified (ADP-free) Hb, as well as hemolysate, resulted in platelet aggregation, as shown by decreased platelet counts. Conclusion: Two primary mechanisms account for RBC hemolysis-associated platelet activation: ADP release, which activates platelets; and cell-free Hb release, which enhances platelet activation by lowering NO bioavailability.

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KW - Nitric oxide

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