Corallo Claudio, Paulesu Luana, Cutolo Maurizio, Ietta Francesca, Carotenuto Claudio, Mannelli Chiara, Romagnoli Roberta, Nuti Ranuccio, Giordano Nicola
Department of Medicine, Surgery and Neurosciences, University of Siena, Italy.
Department of Life Sciences, University of Siena, Italy.
Clin Exp Rheumatol. 2015 Jul-Aug;33(4 Suppl 91):S98-105. Epub 2015 May 25.
To investigate serum levels, tissue/cellular expression of macrophage migration inhibitory factor (MIF) in patients with limited (lSSc) and diffuse (dSSc) systemic sclerosis.
10 lSSc-patients, 10 dSSc-patients and 10 controls were enrolled. MIF serum levels were assayed by ELISA. MIF and its receptors CD74/CD44 were evaluated by immunohistochemistry on skin biopsies from patients with dSSc, lSSc (affected and not-affected skin) and controls. MIF levels were assessed (ELISA) in supernatants of healthy dermal microvascular endothelial cells (MVECs) and in control (CTR), non-affected SSc (NA) and affected (SSc) fibroblasts treated for 48 h with 10% control serum and 10% SSc-serum. MIF supernatant (ELISA) and mRNA (quantitative real-time PCR) levels were determined in SSc dermal fibroblasts and in control dermal fibroblasts untreated or stimulated at 6 h-24 h-48 h with bleomycin (50 mU/ml).
Serum MIF was significantly higher in dSSc (18.7±4.1 ng/ml, p<0.001) and in lSSc (10.4±4.4 ng/ml, p<0.001) patients respect to controls (2.6±1.4 ng/ml). Enhanced MIF immunoreactivity was found in keratinocytes, fibroblasts, endothelium, sebaceous/sweat glands from lSSc/dSSc affected skin. Faint MIF immunoreactivity was found in control skin and not-affected skin of lSSc patients. No differences were found in CD74/CD44 receptors' analysis among control and dSSc/lSSc affected and non-affected skin. MVECs and fibroblasts (CTR, NA and SSc) produced significantly more MIF, when stimulated with SSc serum respect to control-serum (p<0.001). Finally, MIF mRNA levels significantly increased at 6h (p<0.001) and decreased at 48 h (p<0.001) in control fibroblasts treated with bleomycin compared to control untreated. Simultaneously, MIF supernatant protein levels increased after 48 h (p<0.01) in bleomycin-treated fibroblasts respect to untreated ones.
These results suggest that MIF could be implicated in the pathogenesis of SSc, probably acting as protective factor against the SSc stressful conditions.
研究局限性（lSSc）和弥漫性（dSSc）系统性硬化症患者血清中巨噬细胞移动抑制因子（MIF）水平、组织/细胞表达情况。
纳入10例lSSc患者、10例dSSc患者和10例对照。采用酶联免疫吸附测定（ELISA）法检测MIF血清水平。通过免疫组织化学法对dSSc、lSSc患者（受累及未受累皮肤）及对照的皮肤活检标本进行MIF及其受体CD74/CD44评估。采用ELISA法检测健康真皮微血管内皮细胞（MVECs）上清液以及用10%对照血清和10%SSc血清处理48小时的对照（CTR）、未受累SSc（NA）和成纤维细胞（SSc）中的MIF水平。采用ELISA法和定量实时聚合酶链反应（PCR）法检测未处理或用博来霉素（50 mU/ml）在6小时、24小时、48小时刺激的SSc真皮成纤维细胞和对照真皮成纤维细胞中的MIF上清液和mRNA水平。
与对照（2.6±1.4 ng/ml）相比，dSSc患者（18.7±4.1 ng/ml，p<0.001）和lSSc患者（10.4±4.4 ng/ml，p<0.001）血清MIF水平显著升高。在lSSc/dSSc受累皮肤的角质形成细胞、成纤维细胞、内皮细胞、皮脂腺/汗腺中发现MIF免疫反应性增强。在对照皮肤和lSSc患者未受累皮肤中发现微弱的MIF免疫反应性。在对照、dSSc/lSSc受累及未受累皮肤的CD
