Purification of Proplatelet Formation (PPF) Stimulating Factor: Thrombin/Antithrombin III Complex Stimulates PPF of Megakaryocytes In Vitro and Platelet Production In Vivo

Yoji Ishida; Kazuo Yano; Toshiharu Ito; Hiroki Shigematus; Kohsuke Sasaki; Shuhei Kondo; Shin-Ichiro Kuriya

doi:10.1055/s-0037-1615691

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2001; 85(02): 349-355
DOI: 10.1055/s-0037-1615691

Review Article

Schattauer GmbH

Purification of Proplatelet Formation (PPF) Stimulating Factor: Thrombin/Antithrombin III Complex Stimulates PPF of Megakaryocytes In Vitro and Platelet Production In Vivo

Yoji Ishida

¹Division of Hematology, IIIrd Department of Internal Medicine, Iwate Medical University School of Medicine, Morioka, Iwate, Japan

,

Kazuo Yano

²Bio-Science Laboratory, Life Science Research Laboratories, Asahi Chemical Industry, Fuji, Shizuoka, Japan

,

Toshiharu Ito

¹Division of Hematology, IIIrd Department of Internal Medicine, Iwate Medical University School of Medicine, Morioka, Iwate, Japan

,

Hiroki Shigematus

²Bio-Science Laboratory, Life Science Research Laboratories, Asahi Chemical Industry, Fuji, Shizuoka, Japan

,

Kohsuke Sasaki

³Department of Pathology, Yamaguchi University School of Medicine, Ube, Yamaguchi, Japan

,

Shuhei Kondo

²Bio-Science Laboratory, Life Science Research Laboratories, Asahi Chemical Industry, Fuji, Shizuoka, Japan

,

Shin-Ichiro Kuriya

¹Division of Hematology, IIIrd Department of Internal Medicine, Iwate Medical University School of Medicine, Morioka, Iwate, Japan

› Author Affiliations

Abstract

Summary

In this study, the protein which stimulates proplatelet formation (PPF) of megakaryocytes was purified from normal human plasma using 7 steps procedures. Two different protease inhibitors were identified based on their amino acid sequences, i.e. antithrombin III (AT III) and C1 inhibitor. They were included in high density lipoprotein (HDL). HDL was necessary for AT III to be active in PPF in vitro. The biological effects of the AT III /HDL or thrombin-AT III (TAT)/HDL were studied in vitro. PPF of murine megakaryocytes was stimulated by negative control (BSA) (1.8 ± 0.3%), AT III (2.0 ± 0.4%), HDL (1.2 ± 0.9%), AT III /HDL (14.8 ± 2.1%) or TAT/HDL (23.3 ± 3.5%), respectively. TAT/HDL also had a synergistic effect with the mpl ligand, judging by the acetylcholinesterase (AchE) expression of murine megakaryocytes (2.7 fold increase). In vivo subcutaneous administration of AT III alone or TAT for 3 days significantly stimulated thrombocytosis (136% and 144%, respectively, p <0.05) and AT III/HDL showed rapid and further stimulation (150%, p <0.01). These results and the previous studies indicate that megakaryocytopoiesis is regulated by the mpl ligand, while a protease/protease inhibitor complex such as TAT, which is involved in the coagulation cascade associated with platelet consumption, might be one of the regulators in platelet production.

Keywords

Thrombocytopoiesis - proplatelet formation stimulating factor - anti-thrombin III - thrombin - high density lipoprotein

Full Text

References

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