Leukocyte- and platelet-derived microparticles correlate with thrombus weight and tissue factor activity in an experimental mouse model of venous thrombosis

Eduardo Ramacciotti; Angela E. Hawley; Diana M. Farris; Nicole E. Ballard; Shirley K. Wrobleski; Daniel D. Myers Jr; Peter K. Henke; Thomas W. Wakefield

doi:10.1160/TH08-09-0620

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2009; 101(04): 748-754
DOI: 10.1160/TH08-09-0620

Animal Models

Schattauer GmbH

Leukocyte- and platelet-derived microparticles correlate with thrombus weight and tissue factor activity in an experimental mouse model of venous thrombosis

Eduardo Ramacciotti

¹Conrad Jobst Vascular Research Laboratories, Section of Vascular Surgery, University of Michigan Medical Center, Ann Arbor, Michigan, USA

,

Angela E. Hawley

¹Conrad Jobst Vascular Research Laboratories, Section of Vascular Surgery, University of Michigan Medical Center, Ann Arbor, Michigan, USA

,

Diana M. Farris

¹Conrad Jobst Vascular Research Laboratories, Section of Vascular Surgery, University of Michigan Medical Center, Ann Arbor, Michigan, USA

,

Nicole E. Ballard

¹Conrad Jobst Vascular Research Laboratories, Section of Vascular Surgery, University of Michigan Medical Center, Ann Arbor, Michigan, USA

,

Shirley K. Wrobleski

¹Conrad Jobst Vascular Research Laboratories, Section of Vascular Surgery, University of Michigan Medical Center, Ann Arbor, Michigan, USA

,

Daniel D. Myers Jr

¹Conrad Jobst Vascular Research Laboratories, Section of Vascular Surgery, University of Michigan Medical Center, Ann Arbor, Michigan, USA

²Unit for Laboratory Animal Medicine, University of Michigan Medical Center, Ann Arbor, Michigan, USA

,

Peter K. Henke

¹Conrad Jobst Vascular Research Laboratories, Section of Vascular Surgery, University of Michigan Medical Center, Ann Arbor, Michigan, USA

,

Thomas W. Wakefield

¹Conrad Jobst Vascular Research Laboratories, Section of Vascular Surgery, University of Michigan Medical Center, Ann Arbor, Michigan, USA

› Author Affiliations

Abstract

Summary

Microparticles (MP) are lipid vesicles from platelets, leukocytes and endothelial cells that are involved in early thrombogenesis. We evaluated a detailed time-course analysis of MPs on thrombogenesis and the associated tissue factor (TF) activity in wild-type, in gene-deleted for E- and P-selectins and with high levels of P-selectin expression after the initiation of venous thrombosis in mice.Inferior vena cava (IVC) ligation was performed on C57BL/6 mice (n =191, 59 = wild-type [WT], 55 = gene-deleted for E- and P – selectins [knock-outs, EPKO] and 77 = elevated levels of soluble P-selectin, named Delta Cytoplasmic Tail (ΔCT). Animals were euthanised at various time points to assess MP production, origin and thrombus weight. MPs were re-injected into separate mice at concentrations of 80,000 and 160,000 units, as well as from different ages. In addition, MPs from thrombosed animals were pooled and TF activity quantitated using a chromogenic assay. Thrombus weight correlated negatively with MPs derived from leukocytes, and positively with MPs derived from platelets for WT animals (p<0.05), while MPs from platelets presented a positive correlation to thrombus weight in the WT and EPKO groups (p<0.01). Total MPs correlated negatively with thrombus weight in the ΔCT group (p<0.05). MP re-injections led to greater thrombus weight, while older MP reinjections tended to form larger thrombus than younger. Finally, TF bearing MPs showed a significant correlation to MP concentrations (R=0.99). In conclusion, MPs appear to be an important element in venous thrombogenesis.

Keywords

Venous thrombosis - inflammation - microparticles - plasminogen - tissue factor - tissue factor activity - animal models

Full Text

References

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