92-kD gelatinase is released by the prolonged survival neutrophils in gingival crevicular fluid and cleaves the extracellar domain of recombinant 180-kD bullous pemphigoid autoantigen.

本报告介绍了实验的结果，旨在研究的可能性，即中性粒细胞暴露于已知的调节炎症反应的药物可以影响中性粒细胞的生存和干扰这些细胞的凋亡的生理过程。此外，我们研究了中性粒细胞是否是92 kD明胶酶的可能来源，这种酶切割半桥粒元件，重组180 kD大疱性类天疱疮（BP）自身抗原的细胞外结构域，表皮半桥粒的跨膜分子。培养72小时后，100 ng/mL LPS处理的嗜中性粒细胞显示出96.7%的存活率（]SY+-[）10.5%，并且当嗜中性粒细胞用GCF样品处理时，在72小时，AP衍生的GCF的存活百分比为42.7%（]SY+-[）3.8%。根据GCF明胶酶在SDS-PAGE上的迁移模式和APMA活化的影响，鉴定了主要的GCF酶是与来自溶解的中性粒细胞的明胶酶一起迁移的潜伏的92-kD明胶酶，以及更快迁移的活化形式。牙周炎患者的总明胶酶活性（活性加潜伏性）水平高于对照组患者（AP的明胶酶活性中位数为1843单位，EOP为2241单位，对照组为452单位）。此外，评估如果92 kD明胶酶有助于半桥粒元素的破坏，我们孵育纯化，活化酶与42 kD的重组融合蛋白，包括大部分的第一和最大的胶原结构域。基于约8 kD的减少，92 kD明胶酶在胶原胞外结构域的开始切割重组BP 180，并且对于明胶，将胶原序列降解成太小而不能在凝胶上看到的片段。总之，我们的研究结果表明，92 kD明胶酶的完整的衰老中性粒细胞在GCF中的释放可能是一个关键的事件口袋的形成和牙周病的发展。

This report presents the results of experiments designed to examine the possibility that the exposure of neutrophils to agents known to regulate inflammatory reactions can influence neutrophils survival and interfere with the physiologic process of apoptosis these cells. Moreovere, we examined whether neutrophils were a possible source of the 92-kD gelatinase and this enzyme cleaved hemidesmosomal elements, the extracelular domain of recombinant 180-kD bullous pemphigoid (BP) autoantigen, a transmembrane molecule of the epidermal hemidesmosome. After 72 hrs of culture, 100 ng/mL LPS treated neutrophils showed a survival of 96.7%(]SY+-[)10.5%, and also when neutrophils were treated with GCF samples, at 72 hrs the survival percentages were for AP-derived GCF 42.7%(]SY+-[)3.8%. Based on the migration patterns of GCF gelatinase on SDS-PAGE and the effects of APMA activation, the major GCF enzymes were identified as the latent 92-kD progelatinase that comigrated with progelatinase from lysed neutrophils, and the faster migrating activated forms. Patients with periodontitis showed higher levels of total (active plus latent) gelatinase activity than control patients (median gelatinase activity of 1843 units for AP,2241 units for EOP and 452 units for controls). Furthermore, to assess if 92-kD gelatinase contributes to the destruction of hemidesmosomal elements, we incubated purified, activated enzyme with a 42-kD recombinant fusion protein, including a large portion of the first and largest collagenous domain. Based on a decrease of -8-kD,92-kD gelatinase cleaved recombinant BP180 at the beginning of the collagenous ectodomain, and as for gelatin, degraded the collagenous sequences into fragments too small to be seen on the gel. In conclusion, our findings suggest that the release of 92-kD gelatinase by intact senescent neutrophils in GCF may be a critical event in pocket formation and its development seen in periodontal disease.