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

Joellen H H Lin, Mathieu Garand, Branislava Zagorac, Steven L. Schadinger, Corey Scipione, Marlys L. Koschinsky, Michael B. Boffa

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)999-1009
Number of pages11
JournalThrombosis and Haemostasis
Volume105
Issue number6
DOIs
Publication statusPublished - 1 Jun 2011
Externally publishedYes

Fingerprint

Carboxypeptidase B2
Forensic Anthropology
Blood Vessels
Cell Line
Messenger RNA
Endothelial Cells
Macrophages
Carboxypeptidases
Polymerase Chain Reaction
Umbilical Veins
Enzyme Precursors
Umbilical Arteries
Hep G2 Cells
Conditioned Culture Medium
Vascular Diseases
Cell Differentiation
Hepatocellular Carcinoma

Keywords

  • Gene expression
  • Megakaryocytes
  • Monocytes/macrophages
  • Platelets
  • Thrombin-activatable fibrinolysis inhibitor (TAFI)

ASJC Scopus subject areas

  • Hematology

Cite this

Lin, J. H. H., Garand, M., Zagorac, B., Schadinger, S. L., Scipione, C., Koschinsky, M. L., & Boffa, M. B. (2011). Identification of human thrombin-activatable fibrinolysis inhibitor in vascular and inflammatory cells. Thrombosis and Haemostasis, 105(6), 999-1009. https://doi.org/10.1160/TH10-06-0413

Identification of human thrombin-activatable fibrinolysis inhibitor in vascular and inflammatory cells. / Lin, Joellen H H; Garand, Mathieu; Zagorac, Branislava; Schadinger, Steven L.; Scipione, Corey; Koschinsky, Marlys L.; Boffa, Michael B.

In: Thrombosis and Haemostasis, Vol. 105, No. 6, 01.06.2011, p. 999-1009.

Research output: Contribution to journalArticle

Lin, JHH, Garand, M, Zagorac, B, Schadinger, SL, Scipione, C, Koschinsky, ML & Boffa, MB 2011, 'Identification of human thrombin-activatable fibrinolysis inhibitor in vascular and inflammatory cells', Thrombosis and Haemostasis, vol. 105, no. 6, pp. 999-1009. https://doi.org/10.1160/TH10-06-0413
Lin, Joellen H H ; Garand, Mathieu ; Zagorac, Branislava ; Schadinger, Steven L. ; Scipione, Corey ; Koschinsky, Marlys L. ; Boffa, Michael B. / Identification of human thrombin-activatable fibrinolysis inhibitor in vascular and inflammatory cells. In: Thrombosis and Haemostasis. 2011 ; Vol. 105, No. 6. pp. 999-1009.
@article{88d5d4fec8ac4eb98e506d18ed1c7d13,
title = "Identification of human thrombin-activatable fibrinolysis inhibitor in vascular and inflammatory cells",
abstract = "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 = "Gene expression, Megakaryocytes, Monocytes/macrophages, Platelets, Thrombin-activatable fibrinolysis inhibitor (TAFI)",
author = "Lin, {Joellen H H} and Mathieu Garand and Branislava Zagorac and Schadinger, {Steven L.} and Corey Scipione and Koschinsky, {Marlys L.} and Boffa, {Michael B.}",
year = "2011",
month = "6",
day = "1",
doi = "10.1160/TH10-06-0413",
language = "English",
volume = "105",
pages = "999--1009",
journal = "Thrombosis and Haemostasis",
issn = "0340-6245",
publisher = "Schattauer GmbH",
number = "6",

}

TY - JOUR

T1 - Identification of human thrombin-activatable fibrinolysis inhibitor in vascular and inflammatory cells

AU - Lin, Joellen H H

AU - Garand, Mathieu

AU - Zagorac, Branislava

AU - Schadinger, Steven L.

AU - Scipione, Corey

AU - Koschinsky, Marlys L.

AU - Boffa, Michael B.

PY - 2011/6/1

Y1 - 2011/6/1

N2 - 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.

AB - 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.

KW - Gene expression

KW - Megakaryocytes

KW - Monocytes/macrophages

KW - Platelets

KW - Thrombin-activatable fibrinolysis inhibitor (TAFI)

UR - http://www.scopus.com/inward/record.url?scp=79958177151&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=79958177151&partnerID=8YFLogxK

U2 - 10.1160/TH10-06-0413

DO - 10.1160/TH10-06-0413

M3 - Article

C2 - 21505719

AN - SCOPUS:79958177151

VL - 105

SP - 999

EP - 1009

JO - Thrombosis and Haemostasis

JF - Thrombosis and Haemostasis

SN - 0340-6245

IS - 6

ER -