Endotoxaemia-augmented murine venous thrombosis is dependent on TLR-4 and ICAM-1, and potentiated by neutropenia

Andrea T. Obi; Elizabeth Andraska; Yogendra Kanthi; Chase W. Kessinger; Megan Elfline; Cathy Luke; Teruna J. Siahaan; Farouc A. Jaffer; Thomas W. Wakefield; Peter K. Henke

doi:10.1160/TH16-03-0218

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2017; 117(02): 339-348
DOI: 10.1160/TH16-03-0218

Blood Cells, Inflammation and Infection

Schattauer GmbH

Endotoxaemia-augmented murine venous thrombosis is dependent on TLR-4 and ICAM-1, and potentiated by neutropenia^[*]

Andrea T. Obi

¹Conrad Jobst Vascular Research Laboratory, Section of Vascular Surgery, Department of Surgery, University of Michigan, Ann Arbor, Michigan, USA

,

Elizabeth Andraska

¹Conrad Jobst Vascular Research Laboratory, Section of Vascular Surgery, Department of Surgery, University of Michigan, Ann Arbor, Michigan, USA

²University of Michigan Medical School, Ann Arbor, Michigan, USA

,

Yogendra Kanthi

³Department of Cardiology, University of Michigan, Ann Arbor, Michigan, USA

,

Chase W. Kessinger

⁴Cardiovascular Research Center, Cardiology Division, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, USA

,

Megan Elfline

¹Conrad Jobst Vascular Research Laboratory, Section of Vascular Surgery, Department of Surgery, University of Michigan, Ann Arbor, Michigan, USA

,

Cathy Luke

¹Conrad Jobst Vascular Research Laboratory, Section of Vascular Surgery, Department of Surgery, University of Michigan, Ann Arbor, Michigan, USA

,

Teruna J. Siahaan

⁵School of Pharmacology, University of Kansas, Lawrence, Kansas, USA

,

Farouc A. Jaffer

⁴Cardiovascular Research Center, Cardiology Division, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, USA

,

Thomas W. Wakefield

¹Conrad Jobst Vascular Research Laboratory, Section of Vascular Surgery, Department of Surgery, University of Michigan, Ann Arbor, Michigan, USA

,

Peter K. Henke

¹Conrad Jobst Vascular Research Laboratory, Section of Vascular Surgery, Department of Surgery, University of Michigan, Ann Arbor, Michigan, USA

› Author Affiliations

Abstract

Summary

Venous thromboembolism is a major cause of death during and immediately post-sepsis. Venous thrombosis (VT) is mediated by cell adhesion molecules and leukocytes, including neutrophil extracellular traps (NETs). Sepsis, or experimentally, endotoxaemia, shares similar characteristics and is modulated via toll like receptor 4 (TLR4). This study was undertaken to determine if endotoxaemia potentiates early stasis thrombogenesis, and secondarily to determine the role of VT TLR4, ICAM-1 and neutrophils (PMNs). Wild-type (WT), ICAM-1^-/- and TLR4^-/- mice underwent treatment with saline or LPS (10 mg/kg i.p.) alone, or followed by inferior vena cava (IVC) ligation to generate stasis VT. In vivo microscopy of leukocyte trafficking was performed in non-thrombosed mice, and tissue and plasma were harvested during early VT formation. Pre-thrombosis, circulating ICAM-1 was elevated and increased leukocyte adhesion and rolling occurred on the IVC of LPS-treated mice. Post-thrombosis, endotoxaemic mice formed larger, platelet-poor thrombi. Endotoxaemic TLR4^-/- mice did not have an augmented thrombotic response and exhibited significantly decreased circulating ICAM-1 compared to endotoxaemic WT controls. Endotoxaemic ICAM-1^-/- mice had significantly smaller thrombi compared to controls. Hypothesising that PMNs localised to the inflamed endothelium were promoting thrombosis, PMN depletion using anti-Ly6G antibody was performed. Paradoxically, VT formed without PMNs was amplified, potentially related to endotoxaemia induced elevation of PAI-1 and circulating FXIII, and decreased uPA. Endotoxaemia enhanced early VT occurs in a TLR-4 and ICAM-1 dependent fashion, and is potentiated by neutropenia. ICAM-1 and/or TLR-4 inhibition may be a unique strategy to prevent sepsis-associated VT.

Supplementary Material to this article is available online at www.thrombosis-online.com.

Keywords

TLR4 - cellular adhesion molecules - ICAM-1 - sepsis - LPS - venous thrombosis - venous thromboembolism

Full Text

References

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Supplementary Material

Online Supplementary Material (PDF)

Endotoxaemia-augmented murine venous thrombosis is dependent on TLR-4 and ICAM-1, and potentiated by neutropenia[*]

Endotoxaemia-augmented murine venous thrombosis is dependent on TLR-4 and ICAM-1, and potentiated by neutropenia^[*]