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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, and GUIDO ROSSI
ABSTRACT.
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. (J Rheumatol 2005;32:656-64) Key Indexing Terms:
GRANULOCYTE-MACROPHAGE COLONY STIMULATING FACTOR
From the Dipartimento di Biologia e Patologia Cellulare e Molecolare, Dipartimento di Scienze Biomorfologiche e Funzionali, Dipartimento di Medicina Clinica e Sperimentale, Università degli Studi "Federico II," Naples, Italy. Supported by grant L. 41 (1999) from Council of Regione Campania. L. Postiglione, PhD; P. Ladogana, MD, Dipartimento di Biologia e Patologia Cellulare e Molecolare; S. Montagnani, MD, Dipartimento di Scienze Biomorfologiche e Funzionali; G. Di Spigna, PhD, Dipartimento di Biologia e Patologia Cellulare e Molecolare; C. Castaldo, MD, Dipartimento di Scienze Biomorfologiche e Funzionali; M. Turano, PhD, Consorzio Biogem, Naples; E.M. Bruno, PhD, Dipartimento di Biologia e Patologia Cellulare e Molecolare; F. Di Meglio, MD, Dipartimento di Scienze Biomorfologiche e Funzionali; A. Riccio, PhD, Dip. di Medicina Clinica e Sperimentale; G. Rossi, MD, Dipartimento di Biologia e Patologia Cellulare e Molecolare. Address reprint requests to L. Postiglione, Department of Cellular and Molecular Biology and Pathology, Federico II University, Via S. Pansini 4, Ed. 19, 80131 Naples, Italy. E-mail: lorposti@unina.it Accepted for publication December 20, 2004. |