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Open Access Research article

Neutrophil-derived microparticles induce myeloperoxidase-mediated damage of vascular endothelial cells

Thassila Nogueira Pitanga12, Luciana de Aragão França1, Viviane Costa Junqueira Rocha1, Thayna Meirelles3, Valéria Matos Borges1, Marilda Souza Gonçalves1, Lain Carlos Pontes-de-Carvalho1, Alberto Augusto Noronha-Dutra14 and Washington Luis Conrado dos-Santos1*

Author Affiliations

1 Fundação Oswaldo Cruz-BA, Gonçalo Moniz Research Center, Salvador, BA, Brazil

2 Health Science Institute, Federal University of Bahia, Salvador, BA, Brazil

3 Heart Institute, Faculty of Medicine, USP, São Paulo, SP, Brazil

4 University College London, Gower Street, London, UK

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BMC Cell Biology 2014, 15:21  doi:10.1186/1471-2121-15-21

Published: 11 June 2014

Abstract

Background

Upon activation neutrophil releases microparticles - small plasma membrane vesicles that contain cell surface proteins and cytoplasmic matter, with biological activities. In this study we investigated the potential role of myeloperoxidase in the endothelial cell injury caused by neutrophil-derived microparticles.

Results

Microparticles were produced by activating human neutrophils with a calcium ionophore and characterized by flow cytometry and transmission and scanning electron microscopy. Myeloperoxidase activity was measured by luminol-dependent chemiluminescence. Neutrophil microparticles-induced injuries and morphological alterations in human umbilical vein endothelial cells (HUVECs) were evaluated by microscopy and flow cytometry. Neutrophil microparticles were characterized as structures bounded by lipid bilayers and were less than 1 μm in diameter. The microparticles also expressed CD66b, CD62L and myeloperoxidase, which are all commonly expressed on the surface of neutrophils, as well as exposition of phosphatidylserine. The activity of the myeloperoxidase present on the microparticles was confirmed by hypochlorous acid detection. This compound is only catalyzed by myeloperoxidase in the presence of hydrogen peroxide and chloride ion. The addition of sodium azide or taurine inhibited and reduced enzymatic activity, respectively. Exposure of HUVEC to neutrophil microparticles induced a loss of cell membrane integrity and morphological changes. The addition of sodium azide or myeloperoxidase-specific inhibitor-I consistently reduced the injury to the endothelial cells. Taurine addition reduced HUVEC morphological changes.

Conclusions

We have demonstrated the presence of active myeloperoxidase in neutrophil microparticles and that the microparticle-associated myeloperoxidase cause injury to endothelial cells. Hence, the microparticle-associated myeloperoxidase-hydrogen peroxide-chloride system may contribute to widespread endothelial cell damage in conditions of neutrophil activation as observed in vasculitis and sepsis.

Keywords:
Neutrophil microparticles; Endothelial cell injury; Myeloperoxidase; Oxidative stress