Role of Monocyte Chemoattractant Protein-1 and its Receptor,CCR-2, in the Pathogenesis of Bleomycin-Induced Scleroderma Toshiyuki Yamamoto, Kiyoshi Nishioka

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Presentation transcript:

Role of Monocyte Chemoattractant Protein-1 and its Receptor,CCR-2, in the Pathogenesis of Bleomycin-Induced Scleroderma Toshiyuki Yamamoto, Kiyoshi Nishioka Journal of Investigative Dermatology Volume 121, Issue 3, Pages 510-516 (September 2003) DOI: 10.1046/j.1523-1747.2003.12408.x Copyright © 2003 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 Immunohistologic detection of MCP-1 and CCR-2 in the lesional skin following treatment with either PBS (4 wk) or bleomycin (BLM; 1–4 wk). MCP-1 expression was faint to absent in mononuclear cells in the PBS-treated skin (A). By contrast, MCP-1 expression is upregulated on the infiltrating mononuclear cells in the lower dermis at 2 wk (B), and furthermore on fibroblastic cells (arrowheads) in the sclerotic skin at 4 wk (C). CCR-2 expression was weakly detected in the PBS-treated skin (D), whereas increased on the infiltrating mononuclear cells (E) as well as on fibroblastic cells (arrowheads) (F) in the bleomycin-treated skin. Magnification: (A,B,D,E)×á200; (C,F)×á400. Scale bar: (A,B,D,E) 50 μm; (C,F) 25 μm. Journal of Investigative Dermatology 2003 121, 510-516DOI: (10.1046/j.1523-1747.2003.12408.x) Copyright © 2003 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Reverse transcriptase–PCR analysis of MCP-1, MIP-1α, MIP-1β, and RANTES. Total RNA was isolated from skin samples treated with either PBS for 4 wk (Cont) or bleomycin (1–4 wk), and analyzed with specific primers. Results shown are representative of three independent experiments. Journal of Investigative Dermatology 2003 121, 510-516DOI: (10.1046/j.1523-1747.2003.12408.x) Copyright © 2003 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 Reverse transcriptase–PCR analysis of CCR-1 and CCR-2. Total RNA was isolated from skin samples treated with either PBS for 4 wk (Cont) or bleomycin (1–4 wk), and analyzed with specific primers. Results shown are representative of three independent experiments. Journal of Investigative Dermatology 2003 121, 510-516DOI: (10.1046/j.1523-1747.2003.12408.x) Copyright © 2003 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 4 Effect of anti-MCP-1 antibody on bleomycin-induced dermal sclerosis. (A) Anti-MCP-1 antibody blocks the induction of scleroderma induced by bleomycin. Mice were treated with bleomycin (1 mg per mL) together with intravenous administration of either anti-MCP-1 antibody (n=6) or NGS (n=6). Mice treated with bleomycin and NGS for 4 wk demonstrate definite dermal sclerosis with thickened collagen bundles in the thickened dermis (upper). Mice treated with bleomycin and anti-MCP-1 neutralizing antibody for 4 wk show reduction of dermal sclerosis (lower). Hematoxylin and eosin stain, original magnification×á140. Scale bar: 100 μm. (B) Type I collagen mRNA expression. Total RNA was isolated from the lesional skin and analyzed by reverse transcriptase–PCR. Lane 1: mice treated with bleomycin and NGS for 4 wk. Lane 2: anti-MCP-1 treated mice together with bleomycin for 4 wk. Results shown are representative of three independent experiments. (C) Collagen content in the skin. Each value (% of control) was shown as closed circles. Mean±SD are shown. Collagen content was significantly suppressed in mice treated with intravenous administration of anti-MCP-1 antibody together with local bleomycin, compared with those treated with NGS and bleomycin (p<0.05). Journal of Investigative Dermatology 2003 121, 510-516DOI: (10.1046/j.1523-1747.2003.12408.x) Copyright © 2003 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 5 Effect of MCP-1 on α1(I) collagen, fibronectin, decorin, and biglycan mRNA expression in human skin fibroblasts. Semiconfluent cultures of normal dermal fibroblasts were incubated for various time periods (6–24 h) in the presence of 10 ng per mL MCP-1 in serum-free medium. Total RNA (5 μg per lane) was analyzed by northern blot hybridization with cDNA probes, as described in Materials and Methods. As a control, RNA from a parallel culture (unstimulated), which was maintained for 24 h without MCP-1, was loaded. Results shown are representative of three independent experiments. Journal of Investigative Dermatology 2003 121, 510-516DOI: (10.1046/j.1523-1747.2003.12408.x) Copyright © 2003 The Society for Investigative Dermatology, Inc Terms and Conditions