Tomimura Saori, Ogawa Fumihide, Iwata Yohei, Komura Kazuhiro, Hara Toshihide, Muroi Eiji, Takenaka Motoi, Shimizu Kazuhiro, Hasegawa Minoru, Fujimoto Manabu, Sato Shinichi
Department of Dermatology, Nagasaki University Graduate School of Biomedical Sciences, 1-7-1 Sakamoto, Nagasaki, Japan.
J Dermatol Sci. 2008 Oct;52(1):47-54. doi: 10.1016/j.jdermsci.2008.04.013. Epub 2008 Jun 18.
Localized scleroderma (LSc) is characterized by cutaneous fibrosis and various autoantibodies.
To determine the presence or levels of antibodies (Abs) against matrix metalloproteinase (MMP)-1 and their clinical relevance in LSc.
Anti-MMP-1 Ab was examined by ELISA (Enzyme-Linked ImmunoSorbent Assay) and immunoblotting using human recombinant MMP-1. MMP-1 collagenase activity was determined using biotinylated collagen as substrate and the amount of cleaved biotinylated fragments of collagen by MMP-1 was measured by ELISA.
LSc patients exhibited significantly elevated IgG anti-MMP-1 Ab levels relative to normal controls at similar level of patients with systemic sclerosis (SSc). However, IgG anti-MMP-1 Ab levels were comparable among the 3 LSc subgroups: morphea, linear scleroderma, and generalized morphea. When absorbance values higher than the mean+2S.D. of normal controls were considered positive, IgG or IgM anti-MMP-1 Ab was found in 46% and 49% of total LSc patients and SSc patients, respectively. Anti-MMP-1 Ab was detected most frequently in morphea patients (60%), followed by linear scleroderma patients (47%) and then generalized morphea patients (25%). LSc patients positive for IgG anti-MMP-1 Ab had elevated levels of IgG anti-single-stranded DNA Ab, IgG anti-nucleosome Ab, and shorter disease duration relative to those negative. The presence of anti-MMP-1 Ab in LSc patients was confirmed by immunoblotting. IgG isolated from LSc patients' sera positive for IgG anti-MMP-1 Ab by ELISA inhibited MMP-1 collagenase activity.
These results suggest that anti-MMP-1 autoantibody is a novel autoantibody in LSc.
局限性硬皮病(LSc)的特征是皮肤纤维化和多种自身抗体。
确定抗基质金属蛋白酶(MMP)-1抗体(Abs)的存在情况或水平及其在局限性硬皮病中的临床相关性。
使用人重组MMP-1通过酶联免疫吸附测定(ELISA)和免疫印迹法检测抗MMP-1抗体。以生物素化胶原为底物测定MMP-1胶原酶活性,通过ELISA测定MMP-1切割生物素化胶原片段的量。
与正常对照相比,局限性硬皮病患者的IgG抗MMP-1抗体水平显著升高,与系统性硬化症(SSc)患者处于相似水平。然而,在局限性硬皮病的3个亚组(硬斑病、线状硬皮病和泛发性硬斑病)中,IgG抗MMP-1抗体水平相当。当吸光度值高于正常对照平均值+2个标准差时被视为阳性,分别在46%的局限性硬皮病患者和49%的系统性硬化症患者中发现IgG或IgM抗MMP-1抗体。抗MMP-1抗体在硬斑病患者中检出率最高(60%),其次是线状硬皮病患者(47%),然后是泛发性硬斑病患者(25%)。IgG抗MMP-1抗体阳性的局限性硬皮病患者相对于阴性患者,其IgG抗单链DNA抗体、IgG抗核小体抗体水平升高且病程较短。通过免疫印迹法证实了局限性硬皮病患者中抗MMP-1抗体的存在。从ELISA检测IgG抗MMP-1抗体阳性的局限性硬皮病患者血清中分离出的IgG可抑制MMP-1胶原酶活性。
这些结果表明抗MMP-1自身抗体是局限性硬皮病中的一种新型自身抗体。