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01.12.2017 | Research article | Ausgabe 1/2017 Open Access

Arthritis Research & Therapy 1/2017

Increased serum levels of fractalkine and mobilisation of CD34+CD45 endothelial progenitor cells in systemic sclerosis

Arthritis Research & Therapy > Ausgabe 1/2017
Audrey Benyamine, Jérémy Magalon, Sylvie Cointe, Romaric Lacroix, Laurent Arnaud, Nathalie Bardin, Pascal Rossi, Yves Francès, Fanny Bernard-Guervilly, Gilles Kaplanski, Jean-Robert Harlé, Pierre-Jean Weiller, Philippe Berbis, David Braunstein, Elisabeth Jouve, Nathalie Lesavre, Françoise Couranjou, Françoise Dignat-George, Florence Sabatier, Pascale Paul, Brigitte Granel
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Electronic supplementary material

The online version of this article (doi:10.​1186/​s13075-017-1271-7) contains supplementary material, which is available to authorized users.



The disruption of endothelial homeostasis is a major determinant in the pathogenesis of systemic sclerosis (SSc) and is reflected by soluble and cellular markers of activation, injury and repair. We aimed to provide a combined assessment of endothelial markers to delineate specific profiles associated with SSc disease and its severity.


We conducted an observational, single-centre study comprising 45 patients with SSc and 41 healthy control subjects. Flow cytometry was used to quantify circulating endothelial microparticles (EMPs) and CD34+ progenitor cell subsets. Colony-forming unit-endothelial cells (CFU-ECs) were counted by culture assay. Circulating endothelial cells were enumerated using anti-CD146-based immunomagnetic separation. Blood levels of endothelin-1, vascular endothelial growth factor (VEGF) and soluble fractalkine (s-Fractalkine) were evaluated by enzyme-linked immunosorbent assay. Disease-associated markers were identified using univariate, correlation and multivariate analyses.


Enhanced numbers of EMPs, CFU-ECs and non-haematopoietic CD34+CD45 endothelial progenitor cells (EPCs) were observed in patients with SSc. Patients with SSc also displayed higher serum levels of VEGF, endothelin-1 and s-Fractalkine. s-Fractalkine levels positively correlated with CD34+CD45 EPC numbers. EMPs, s-Fractalkine and endothelin-1 were independent factors associated with SSc. Patients with high CD34+CD45 EPC numbers had lower forced vital capacity values. Elevated s-Fractalkine levels were associated with disease severity, a higher frequency of pulmonary fibrosis and altered carbon monoxide diffusion.


This study identifies the mobilisation of CD34+CD45 EPCs and high levels of s-Fractalkine as specific features of SSc-associated vascular activation and disease severity. This signature may provide novel insights linking endothelial inflammation and defective repair processes in the pathogenesis of SSc.
Additional file 1: Table S1. Ongoing treatments in patients with SSc. Table S2. Biological characteristics of the study population. Table S3. Summary of the main immunophenotypic and functional characteristics of the investigated circulating progenitor cell subsets. Table S4. Multivariate logistic regression analysis for SSc with robust estimator. (DOCX 16 kb)
Additional file 2: Figure S1. Gating strategy of endothelial progenitor cells (EPCs) by flow cytometry. CD34+CD45+ haematopoietic progenitor cells (HPCs) and CD34+CD45− EPCs were identified within 7-AAD-negative viable (a), CD34+ cells (b), with CD45dim or CD45− expression (c), and displaying forward scatter (FS)/side scatter (SS) characteristics corresponding to the lymphocyte cluster (d, e). Non-specific binding of CD34-PE antibody is checked on CD34+CD45+ (f) and CD34+CD45− gate of each sample (g) by means of a control tube in which CD34-PE antibody is replaced by its isoclonic control. LYMPH Lymphocytes. Figure S2. CFU-EC count in the peripheral venous blood of healthy control subjects (HC) and patients with systemic sclerosis (SSc). Number (n) of CFU-ECs was determined after cell culture of MNCs. ** P < 0.005. (PDF 311 kb)
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