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PLoS By Category | Recent PLoS Articles
Biochemistry - Dermatology - Immunology - Rheumatology

Differentiation Potential of CD14+ Monocytes into Myofibroblasts in Patients with Systemic Sclerosis
Published: Wednesday, March 14, 2012
Author: Nadine Binai et al.

by Nadine Binai, Steven O'Reilly, Bridget Griffiths, Jacob M. van Laar, Thomas Hügle

Background

Circulating monocytes are a highly plastic and functionally heterogeneic cell type with an activated phenotype in patients with systemic sclerosis (SSc). CD14+ monocytes have the potential to differentiate into extra-cellular matrix (ECM) producing cells, possibly participating in fibrogenesis.

Aim

To study the effect of GM-CSF, IL-4 and endothelin -1 (ET-1) alone or in combination on monocyte differentiation into myofibroblasts.

Methods

CD14+ cells were isolated from peripheral blood from 14 SSc patients and healthy controls by positive selection and incubated with different combinations of GM-CSF, IL-4 and ET-1 for 14 days. Type-1 collagen and a-SMA were detected by Western blot, qPCR and confocal microscopy. HLA-DR, CD11c and CD14 expression was analysed by flow cytometry. A collagen gel contraction assay was performed for functional myofibroblast assessment.

Results

GM-CSF both induced collagen and a-SMA expression after 14 days. ET-1 further increased GM-CSF-induced collagen expression in a dose dependent manner up to 30-fold. IL-4/GM-CSF combination leads to a more DC-like phenotype of monocytes associated with reduced collagen and a-SMA expression compared to GM-CSF alone. Collagen and a-SMA expression was higher in monocytes from SSc patients and monocytes were more prone to obtain a spindle form. In contrast to controls, ET-1 and IL-4 alone were sufficient to induce a-SMA expression in monocytes from SSc patients. Despite the induction of a-SMA expression, monocyte-derived myofibroblasts only had a moderate capability of contraction in functional analyses.

Conclusion

SSc monocytes display increased maturation towards myofibroblasts demonstrated by their phenotype and a-SMA expression when compared to monocytes from healthy controls, however only with minor functional contraction properties.

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