NEUTROPHIL COLLAGENASES - ROLE IN RHEUMATOID ARTHRITIS

胶原蛋白的酶降解被认为在 类风湿性关节炎患者的致残性关节破坏 来自类风湿滑膜细胞的胶原酶已经被 参与了这一过程。中性粒细胞是一种循环细胞，它 聚集在炎症部位，包括类风湿滑膜细胞 在这一过程中受到了牵连。中性粒细胞是一种循环 聚集在包括类风湿在内的炎症部位的细胞 滑液。该细胞至少有三种胶原酶。 酶：一种间质胶原酶，能降解I型、II型和III型 胶原蛋白；降解III型和IV型胶原的弹性淀粉酶；以及 明胶酶，降解天然的V型胶原和变性的 胶原蛋白。这些酶在类风湿疾病中的相对作用 进程处于未知状态。 这项提议的第一个目标是提纯和 牙周炎间质胶原酶和明胶酶的性质 人类中性粒细胞。这将使用标准层析来完成 技术和辅助的额外的单抗。主动者 而这些酶的潜在形式将具有特殊的特征 关注激活过程中发生的变化。 一组针对这些酶的单抗将被提出用于 以下用途：1)作为提纯和 酶的特性；2)确定两者之间的关系 某些酶功能，如底物结合和实际底物 裂解和；以及3)作为检测酶的特异性探针 滑液。后一项目标将包括发展 能识别胶原酶的特异性免疫分析 来源于中性粒细胞。这些化验将被用来量化这些 类风湿滑液中的胶原酶。 类风湿滑液中存在的物质(S)，它能够 激活这些酶的潜在形式将使用 标准的层析技术。这些物质将是 关于它们的分子结构和蛋白水解性 活动。

The enzymatic degradation of collagen is felt to play a major role in the crippling joint destruction seen in patients with rheumatoid arthritis and the collagenase derived from rheumatoid synovial cells has been incriminated in this process. The neutrophil is a circulating cell which accumulates at sites of inflammation including rheumatoid synovial cells has been incriminated in this process. The neutrophil is a circulating cell which accumulates at sites of inflammation including rheumatoid synovial fluid. This cell possesses at least three collagenolytic enzymes: an interstitial collagenase which degrades types I, II, and III collagens; elastase which degrades types III and IV collagens; and a gelatinase which degrades native type V collagen as well as denatured collagens. The relative contribution of these enzymes in the rheumatoid process in unknown. The first objective of this proposal is the purification and characterization of the interstitial collagenase and the gelatinase of the human neutrophil. This will be accomplished using standard chromatographic techniques with the additional aid of monoclonal antibodies. The active and latent forms of these enzymes will be characterized with particular focus on the changes which occur during activation. A panel of monoclonal antibodies will be raised against these enzymes for the following purposes: 1) as an aid in the purification and characterization of the enzymes; 2) to determine the relationship between certain enzyme function such as substrate binding and actual substrate cleavage and; and 3) as specific probes for the detection of the enzymes in synovial fluid. This latter objective will include the development of specific immunoassays which will specifically recognize the collagenases derived from neutrophils. These assays will be used to quantitate these collagenases in rheumatoid synovial fluid. The substance(s) present in rheumatoid synovial fluid which are capable of activating the latent forms of these enzymes will be purified using standard chromatographic techniques. These substances will be characterized with regard to their molecular structure and proteolytic activity.