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Abstract

Effect of granulocyte macrophage-colony stimulating factor on extracellular matrix deposition by dermal fibroblasts from patients with scleroderma.

Loredana Postiglione, Paolo Ladogana, Stefania Montagnani, Gaetano di Spigna, Clotilde Castaldo, Mimmo Turano, Eugenia Maria Bruno, Franca Di Meglio, Antonio Riccio and Guido Rossi
The Journal of Rheumatology April 2005, 32 (4) 656-664;
Loredana Postiglione
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Paolo Ladogana
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Stefania Montagnani
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Gaetano di Spigna
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Clotilde Castaldo
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Mimmo Turano
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Eugenia Maria Bruno
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Franca Di Meglio
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Antonio Riccio
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Guido Rossi
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Abstract

OBJECTIVE:. To investigate the in vitro effect of granulocyte macrophage-colony stimulating factor (GM-CSF) on the deposition of extracellular matrix (ECM) in fibroblasts obtained from the skin of patients with systemic sclerosis (Ssc), compared to healthy controls. METHODS: Dermal fibroblasts obtained from 14 patients with SSc (7 with the diffuse form and 7 with CREST syndrome) and from 7 controls were studied. Both SSc and normal skin fibroblast cultures were stimulated for 4 and 8 days with 100 ng/ml GM-CFS. GM-CSF receptor (GM-CSFR) expression was determined by Western blot of cell lysates. Immunofluorescence was used to determine GM-CSFR expression and to investigate the deposition of ECM (type I collagen, fibronectin, and tenascin). Quantitative analysis of ECM was performed by ELISA. Expression of type I collagen and metalloproteinase 1 (MMP-1) mRNA was determined by real-time quantitative PCR. RESULTS: Deposition of ECM by normal fibroblasts appeared not to be influenced by stimulation with GM-CSF; in contrast, after stimulation with GM-CSF SSc fibroblasts showed increased deposition of fibronectin and tenascin, while type I collagen production was decreased; these results were found with both immunofluorescence and ELISA. Quantitative PCR revealed that GM-CSF inhibited the expression of mRNA type I collagen in SSc fibroblasts but not in normal fibroblasts, whereas levels of the main collagenolytic enzyme, MMP-1, were not affected. CONCLUSION: These results suggest that in SSc fibroblasts GM-CSF exerts a blocking effect on the deposition of type I collagen, through an inhibitory action on mRNA, while the production of other components of ECM such as fibronectin and tenascin is increased by stimulation with this cytokine.

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The Journal of Rheumatology
Vol. 32, Issue 4
1 Apr 2005
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Effect of granulocyte macrophage-colony stimulating factor on extracellular matrix deposition by dermal fibroblasts from patients with scleroderma.
Loredana Postiglione, Paolo Ladogana, Stefania Montagnani, Gaetano di Spigna, Clotilde Castaldo, Mimmo Turano, Eugenia Maria Bruno, Franca Di Meglio, Antonio Riccio, Guido Rossi
The Journal of Rheumatology Apr 2005, 32 (4) 656-664;

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Effect of granulocyte macrophage-colony stimulating factor on extracellular matrix deposition by dermal fibroblasts from patients with scleroderma.
Loredana Postiglione, Paolo Ladogana, Stefania Montagnani, Gaetano di Spigna, Clotilde Castaldo, Mimmo Turano, Eugenia Maria Bruno, Franca Di Meglio, Antonio Riccio, Guido Rossi
The Journal of Rheumatology Apr 2005, 32 (4) 656-664;
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