Mechanism of serum-mediated endothelial injury in scleroderma: Identification of a granular enzyme in scleroderma skin and sera
Mechanism of serum-mediated endothelial injury in scleroderma: Identification of a granular enzyme in scleroderma skin and sera
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DOI：
10.1006/clin.1996.4322
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发表时间：
1997-04-01
期刊：
影响因子：
--
通讯作者：
Fan, PS
中科院分区：
文献类型：
--
作者：
Kahaleh, MB;Fan, PS
硬皮病 (SSc) 血清中描述了分子量为 48-60 kDa 的循环内皮细胞生长抑制因子，并显示出具有蛋白水解作用。鉴于最近在 SSc 中细胞免疫激活的证明，并且由于活化的溶细胞 T 细胞颗粒（颗粒酶）中新型丝氨酸蛋白酶的描述，我们假设颗粒酶代表了 SSc 血清中的内皮抑制原理。从IL-2激活的非贴壁正常淋巴细胞中分离出颗粒酶，并使用苯甲脒亲和柱和分子筛柱分离出60-kDa颗粒酶。用分离的颗粒酶免疫兔子产生多克隆抗血清。抗颗粒酶抗体消除了 SSc 血清介导的 EC 生长抑制。此外，在苯甲脒柱纯化的 SSc 血清的蛋白质印迹上，将与分离的颗粒酶相似的循环蛋白鉴定为 60 kDa 手。使用抗颗粒酶抗体对 SSc 皮肤活检进行的免疫荧光研究表明？存在颗粒酶反应性，而健康对照组织呈阴性。此外，通过 BCR 方法在 SSc 皮肤活检中鉴定了颗粒酶 A 基因表达。这些数据表明细胞溶解机制参与了硬皮病的发病机制。 (C) 1997 年学术出版社。
Circulating endothelial cell growth-inhibitory factor with a molecular weight of 48-60 kDa was described in scleroderma (SSc) sera and shown to have a proteolytic action. In view of the recent demonstration of cellular immune activation in SSc, and because of the description of novel serine proteases in the granules of activated cytolytic T cells (granzymes), we hypothesized that granzymes represent the endothelial inhibitory principal in SSc sera. Granular enzymes were isolated from IL-2-activated nonadherent normal lymphocytes, and a 60-kDa granzyme was isolated using benzamidine-affinity column and molecular sieve column. A polyclonal antiserum was generated by immunizing rabbits with the isolated granzyme. Anti-granzyme antibody abolished SSc serum-mediated EC growth inhibition, Furthermore, a circulating protein similar to isolated granzyme was identified as a 60-kDa hand on Western blots of benzamidine column-purified SSc sera. Immunofluorescence studies of SSc skin biopsies using anti-granzyme antibody demonstrated? the presence of granzyme reactivity, while healthy control tissues were negative. Moreover, granzyme A gene expression was identified in SSc skin biopsies by a BCR method. The data suggest cytolytic mechanism involvement in the pathogenesis of scleroderma. (C) 1997 Academic Press.