Verraes S, Hornebeck W, Polette M, Borradori L, Bernard P
FRE 2260 CNRS, IFR 53 Biomolecules, University of Reims-Champagne Ardennes, Reims, France.
J Invest Dermatol. 2001 Nov;117(5):1091-6. doi: 10.1046/j.0022-202x.2001.01521.x.
Bullous pemphigoid is a blistering disorder associated with autoantibodies directed against two components of hemidesmosomes, BP180 and BP230. Autoantibodies to the extracellular collagenous domain of BP180 are thought to play a key role in the pathogenesis of the disease. In a murine model of bullous pemphigoid, neutrophil elastase and 92 kDa gelatinase (matrix metalloproteinase 9) have been implicated in subepidermal blister formation via proteolytic degradation of BP180. In this study we sought to elucidate the contribution of these two enzymes to subepidermal blister formation by assessing the expression, localization, and activity of the two proteases in lesional skin, serum samples, and blister fluids obtained from 17 patients with bullous pemphigoid. The results indicate that (i) neutrophil elastase is found in skin biopsy specimens from bullous pemphigoid lesions and is recovered as active enzyme in blister fluids, and (ii) although proform of matrix metalloproteinase 9 is present in lesional skin, it is present only as proenzyme in blister fluids, which also contain high levels of tissue inhibitor of metalloproteinase-1. Next, the capacity of matrix metalloproteinase 9 and neutrophil elastase to degrade a recombinant protein corresponding to the extracellular collagenous domain of the BP180 was studied. Our data illustrate that (i) recombinant matrix metalloproteinase 9, neutrophil elastase, and blister fluid from bullous pemphigoid patients are all able to hydrolyze recombinant BP180; (ii) the pattern of recombinant BP180 proteolysis with blister fluid was similar to that obtained with neutrophil elastase; and (iii) recombinant BP180 degradation by blister fluid could be inhibited by chloromethylketone, a specific elastase inhibitor, but not by batimastat, a wide spectrum matrix metalloproteinase inhibitor. Our results confirm the importance of neutrophil elastase but not matrix metalloproteinase 9 in the direct cleavage of BP180 autoantigen and subepidermal blister formation in human bullous pemphigoid.
大疱性类天疱疮是一种水疱性疾病，与针对半桥粒的两种成分BP180和BP230的自身抗体有关。针对BP180细胞外胶原结构域的自身抗体被认为在该疾病的发病机制中起关键作用。在大疱性类天疱疮的小鼠模型中，中性粒细胞弹性蛋白酶和92 kDa明胶酶（基质金属蛋白酶9）通过对BP180的蛋白水解降解参与表皮下疱的形成。在本研究中，我们通过评估17例大疱性类天疱疮患者的皮损、血清样本和疱液中这两种蛋白酶的表达、定位及活性，试图阐明这两种酶对表皮下疱形成的作用。结果表明：（i）在大疱性类天疱疮皮损的皮肤活检标本中发现中性粒细胞弹性蛋白酶，并在疱液中以活性酶形式回收；（ii）虽然基质金属蛋白酶9的前体形式存在于皮损中，但它在疱液中仅以前酶形式存在，疱液中还含有高水平的金属蛋白酶组织抑制剂-1。接下来，研究了基质金属蛋白酶9和中性粒细胞弹性蛋白酶降解对应于BP180细胞外胶原结构域的重组蛋白的能力。我们的数据表明：（i）重组基质金属蛋白酶9、中性粒细胞弹性蛋白酶以及大疱性类天疱疮患者的疱液均能够水解重组BP180；（ii）疱液对重组BP180的蛋白水解模式与中性粒细胞弹性蛋白酶相似；（iii）疱液对重组BP180的降解可被氯甲基酮（一种特异性弹性蛋白酶抑制剂）抑制，但不能被batimastat（一种广谱基质金属蛋白酶抑制剂）抑制。我们的结果证实了中性粒细胞弹性蛋白酶而非基质金属蛋白酶9在人源大疱性类天疱疮中对BP180自身抗原的直接切割及表皮下疱形成中的重要性。
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