Open Access Research article

Systemic sclerosis sera affect fibrillin-1 deposition by dermal blood microvascular endothelial cells: therapeutic implications of cyclophosphamide

Marilisa Villano1, Annalisa Borghini1, Mirko Manetti2*, Erica Gabbrielli1, Antonella Rossi1, Piersante Sestini3, Anna Franca Milia2, Francesca Nacci4, Serena Guiducci4, Marco Matucci-Cerinic4, Lidia Ibba-Manneschi2 and Elisabetta Weber1*

Author Affiliations

1 Department of Molecular and Developmental Medicine, University of Siena, Via Aldo Moro 2, 53100, Siena, Italy

2 Department of Experimental and Clinical Medicine, Section of Anatomy and Histology, University of Florence, Largo Brambilla 3, 50134, Florence, Italy

3 Department of Medicine, Surgery and Neuroscience, University of Siena, Azienda Ospedaliera Universitaria Senese, Viale M Bracci 16, 53100, Siena, Italy

4 Department of Experimental and Clinical Medicine, Section of Internal Medicine, Division of Rheumatology, University of Florence, Azienda Ospedaliero-Universitaria Careggi, Villa Monna Tessa, Viale G Pieraccini 18, 50139, Florence, Italy

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Arthritis Research & Therapy 2013, 15:R90  doi:10.1186/ar4270

Published: 20 August 2013

Abstract

Introduction

Systemic sclerosis (SSc) is a connective tissue disorder characterized by endothelial cell injury, autoimmunity and fibrosis. The following three fibrillin-1 alterations have been reported in SSc. (1) Fibrillin-1 microfibrils are disorganized in SSc dermis. (2) Fibrillin-1 microfibrils produced by SSc fibroblasts are unstable. (3) Mutations in the FBN1 gene and anti-fibrillin-1 autoantibodies have been reported in SSc. Fibrillin-1 microfibrils, which are abundantly produced by blood and lymphatic microvascular endothelial cells (B-MVECs and Ly-MVECs, respectively), sequester in the extracellular matrix the latent form of the potent profibrotic cytokine transforming growth factor β (TGF-β). In the present study, we evaluated the effects of SSc sera on the deposition of fibrillin-1 and microfibril-associated glycoprotein 1 (MAGP-1) and the expression of focal adhesion molecules by dermal B-MVECs and Ly-MVECs.

Methods

Dermal B-MVECs and Ly-MVECs were challenged with sera from SSc patients who were treatment-naïve or under cyclophosphamide (CYC) treatment and with sera from healthy controls. Fibrillin-1/MAGP-1 synthesis and deposition and the expression of αvβ3 integrin/phosphorylated focal adhesion kinase and vinculin/actin were evaluated by immunofluorescence and quantified by morphometric analysis.

Results

Fibrillin-1 and MAGP-1 colocalized in all experimental conditions, forming a honeycomb pattern in B-MVECs and a dense mesh of short segments in Ly-MVECs. In B-MVECs, fibrillin-1/MAGP-1 production and αvβ3 integrin expression significantly decreased upon challenge with sera from naïve SSc patients compared with healthy controls. Upon challenge of B-MVECs with sera from CYC-treated SSc patients, fibrillin-1/MAGP-1 and αvβ3 integrin levels were comparable to those of cells treated with healthy sera. Ly-MVECs challenged with SSc sera did not differ from those treated with healthy control sera in the expression of any of the molecules assayed.

Conclusions

Because of the critical role of fibrillin-1 in sequestering the latent form of TGF-β in the extracellular matrix, its decreased deposition by B-MVECs challenged with SSc sera might contribute to dermal fibrosis. In SSc, CYC treatment might limit fibrosis through the maintenance of physiologic fibrillin-1 synthesis and deposition by B-MVECs.

Keywords:
Systemic sclerosis; blood and lymphatic microvascular endothelial cells; fibrillin-1; focal adhesion molecules; cyclophosphamide