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Open Access Highly Accessed Research article

Increased frequency of Th17 cells in systemic sclerosis is related to disease activity and collagen overproduction

Xiaoqin Yang1, Ji Yang1, Xiaojing Xing1, Linlin Wan2 and Ming Li1*

Author Affiliations

1 Department of Dermatology, Zhongshan Hospital, Fudan University, 180 Fenglin Road, Shanghai 200032, China

2 Department of Dermatology, Shanghai Skin Diseases Hospital, 200 Wuyi Road, Shanghai 200050, China

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Arthritis Research & Therapy 2014, 16:R4  doi:10.1186/ar4430

Published: 7 January 2014

Additional files

Additional file 1: Figure S1:

IL-17 induces fibroblast proliferation and collagen production. (A) Fibroblasts isolated from SSc patients were cultured in 20 ng/ml IL-17 or vehicle for the indicated number of days, and their growth was analyzed by MMT assay. (B) Fibroblasts were cultured in the indicated doses of IL-17 for 48 hours; the gene expression of collagen 1 and collagen 3 was measured by real-time RT-PCR analysis.

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Additional file 2: Figure S2:

IL-17 derived from SSc patients induces collagen gene expression in fibroblasts. (A) Fibroblasts were stimulated with different concentrations of supernatants of PI-stimulated PBMCs from patients with active SSc (Active) and PI-stimulated PBMCs from healthy controls (Control) for 48 hours, and the gene expression of collagen 1 and collagen 3 was measured by real-time RT-PCR analysis. (B) Fibroblasts were stimulated with supernatants of PI-stimulated PBMCs from patients with active SSc (Active) and PI-stimulated PBMCs from healthy controls (Control) for different hours, and the gene expression of collagen 1 and collagen 3 was measured by real-time RT-PCR analysis. Results shown are representative of at least three independent experiments.

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Additional file 3: Figure S3:

ERK activation is involved in the IL-17-mediated fibroblast proliferation and collagen production. (A) Fibroblast was stimulated with indicated doses of IL-17 in the presence or absence of IL-17 neutralizing antibody for 24 hours; collagen-1 and collagen-3 protein expressions in fibroblast were analyzed with Western blot. (B) Fibroblast was stimulated with IL-17 and different doses of ERK- specific phosphorylation inhibitor-PD98059 for 24 hours; collagen-1 and collagen-3 protein expressions in fibroblast were analyzed with Western blot. (C) Fibroblast was stimulated with IL-17 and ERK specific phosphorylation inhibitor-PD98059 for indicated days, the proliferation of fibroblast was examined by cell counting kit-8.

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