Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2011; 105(06): 999-1009
DOI: 10.1160/TH10-06-0413

Blood Coagulation, Fibrinolysis and Cellular Haemostasis

Schattauer GmbH

Identification of human thrombin-activatable fibrinolysis inhibitor in vascular and inflammatory cells

Joellen H. H. Lin

¹Department of Biochemistry, Queen’s University, Kingston, Ontario, Canada

,

Mathieu Garand

²Department of Chemistry and Biochemistry, University of Windsor, Windsor, Ontario, Canada

,

Branislava Zagorac

²Department of Chemistry and Biochemistry, University of Windsor, Windsor, Ontario, Canada

,

Steven L. Schadinger

¹Department of Biochemistry, Queen’s University, Kingston, Ontario, Canada

,

Corey Scipione

²Department of Chemistry and Biochemistry, University of Windsor, Windsor, Ontario, Canada

,

Marlys L. Koschinsky

²Department of Chemistry and Biochemistry, University of Windsor, Windsor, Ontario, Canada

,

Michael B. Boffa

²Department of Chemistry and Biochemistry, University of Windsor, Windsor, Ontario, Canada

› Author Affiliations

Summary

TAFI (thrombin-activatable fibrinolysis inhibitor) is a carboxypeptidase zymogen originally identified in plasma. The TAFI pathway helps to regulate the balance between the coagulation and fibrinolytic cascades. Activated TAFI (TAFIa) can also inactivate certain pro-inflammatory mediators, suggesting that the TAFI pathway may also regulate communication between coagulation and inflammation. Expression in the liver is considered to be the source of plasma TAFI. TAFI has also been identified in platelets and CPB2 (the gene encoding TAFI) mRNA has been detected in megakaryocytic cell lines as well as in endothelial cells. We have undertaken a quantitative analysis of CPB2 mRNA and TAFI protein in extrahepatic cell types relevant to vascular disease. Using RT-PCR and quantitative RT-PCR, we detected CPB2 mRNA in the human megakaryoblastic cell lines MEG-01 and Dami, the human monocytoid cell line THP-1 as well as THP-1 cells differentiated into a macrophage-like phenotype, and in primary human umbilical vein and coronary artery endothelial cells. CPB2 mRNA abundance in MEG-01, Dami, and THP-1 cells was modulated by the state of differentiation of these cells. Using a recently developed TAFIa assay, we detected TAFI protein in the lysates of the human hepatocellular carcinoma cell line HepG2 as well as in MEG-01 and Dami cells and in the conditioned medium of HepG2 cells, differentiated Dami cells, and THP-1 macrophages. We have obtained clear evidence for extrahepatic expression of TAFI, which has clear implications for the physiological and pathophysiological functions of the TAFI pathway.

Keywords

Keywords

Thrombin-activatable fibrinolysis inhibitor (TAFI) - gene expression - megakaryocytes - platelets - monocytes/macrophages

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