Thrombopoietin Contributes to Enhanced Platelet Activation in Patients With Unstable Angina
Enrico Lupia, MD*,
Ornella Bosco, PhD ,
Serena Bergerone, MD ,
Anna Erna Dondi, MD,
Alberto Goffi, MD,
Elena Oliaro, MD,
Marco Cordero, MD,
Lorenzo Del Sorbo, MD,
Giampaolo Trevi, MD and
Giuseppe Montrucchio, MD,*
* Azienda Ospedaliera San Giovanni Battista-Molinette, Turin, Italy
Department of Clinical Pathophysiology, University of Turin, Turin, Italy
Department of Internal Medicine, University of Turin, Turin, Italy.
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Figure 1 (A) Plasma thrombopoietin (TPO) levels in healthy control subjects (C) and in patients with stable angina (SA) and unstable angina (UA). (B) Monocyte-platelet aggregates and (C) platelet P-selectin expression detected in vivo, by flow cytometry, in healthy control subjects and in patients with SA and UA. One-way analysis of variance with Newman-Keuls multicomparison test was performed.
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Figure 2 (A) Representative fluorescence-activated cell sorting (FACS) analysis showing the percentage of c-Mpl expression on platelet surface detected in vivo in healthy control subjects (C) and in patients with stable angina (SA) and unstable angina (UA). The solid curve represents fluorescence due to binding of non-specific antimouse immunoglobulin G monoclonal antibody; the open curve represents fluorescence due to binding of anti–c-Mpl monoclonal antibody. (B) Quantification of c-Mpl expression on platelet surface detected in vivo in healthy controls and in patients with SA and UA by FACS analysis. One-way analysis of variance with Newman-Keuls multicomparison test was performed. (C) Representative Western blot analysis of the phosphorylation state of c-Mpl in platelet lysates from healthy control subjects and from patients with SA and UA. Platelets from control subjects or patients were either unstimulated or stimulated with saturating concentrations of recombinant human thrombopoietin (rhTPO). The c-Mpl was phosphorylated in platelets from SA patients and healthy control subjects but not in platelets from UA patients. Shown is a representative experiment out of 4 with similar results.
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Figure 3 Representative aggregation traces and quantification of the in vitro priming activity induced by plasma from healthy control subjects (C) and patients with stable angina (SA) and unstable angina (UA) on (A) adenosine 5'-diphosphate (ADP)- or (B) epinephrine (EPI)-induced platelet aggregation in PRP. Representative aggregation traces and quantification of the in vitro priming activity induced by plasma from healthy controls and patients with SA and UA on (C) ADP- or (D) EPI-induced platelet aggregation in whole blood. One-way analysis of variance with Newman-Keuls multicomparison test was performed.
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Figure 4 (A) In vitro effect of plasma from healthy control subjects (C) and patients with stable angina (SA) and unstable angina (UA) on adenosine 5'-diphosphate-induced monocyte-platelet aggregation (left) and platelet P-selectin expression (right) in whole blood. (B) In vitro effect of plasma from healthy control subjects and patients with SA and UA on epinephrine-induced monocyte-platelet aggregation (left) and platelet P-selectin expression (right) in whole blood. Monocyte-platelet aggregates and platelet P-selectin expression were analyzed by flow cytometry. Analysis of variance with Newman-Keuls multicomparison test was performed.
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Figure 5 Representative aggregation traces and quantification of the effect of thrombopoietin receptor (TPOR)-Fc chimera on the in vitro priming activity induced by plasma from patients with unstable angina (UA) on (A) adenosine 5'-diphosphate (ADP)- or (B) epinephrine (EPI)-induced platelet aggregation in platelet-rich plasma. Representative aggregation traces and quantification of the effect of TPOR-Fc chimera on the in vitro priming activity induced by plasma from UA patients on (C) ADP- or (D) EPI-induced platelet aggregation in whole blood. One-way analysis of variance with Newman-Keuls multicomparison test was performed.
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Figure 6 (A) Effect of thrombopoietin receptor (TPOR)-Fc chimera on the priming activity induced by plasma from patients with unstable angina (UA) on adenosine 5'-diphosphate (ADP)-induced monocyte-platelet aggregation (left) and platelet P-selectin expression (right) in whole blood. (B) Effect of TPOR-Fc chimera on the priming activity induced by plasma from patients with UA on epinephrine (EPI)-induced monocyte-platelet aggregation (left) and platelet P-selectin expression (right) in whole blood. Pretreatment of plasma sample with the TPOR-Fc chimera significantly reduced the percentages of ADP- or EPI-induced monocyte-platelet binding and platelet P-selectin expression observed after preincubation of whole blood from healthy adult donors with plasma from UA patients. Analysis of variance with Newman-Keuls multicomparison test was performed.
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