NEUTROPHIL COLLAGENASES - ROLE IN RHEUMATOID ARTHRITIS

中性粒细胞胶原酶 - 在类风湿性关节炎中的作用

• 批准号: 3157985
• 负责人: CARLO L. MAINARDI
• 金额: $ 5.85万
• 依托单位: UNIVERSITY OF TENNESSEE HEALTH SCI CTR
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-09-16 至 1988-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3157985
• 关键词: collagenase; elastases; enzyme linked immunosorbent assay; enzyme structure; gel electrophoresis; human tissue; immunologic techniques; monoclonal antibody; neutrophil; radioimmunoassay; rheumatoid arthritis; synovial fluid

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.

胶原蛋白的酶促降解被认为在胶原蛋白的降解中起主要作用。 类风湿性关节炎患者的关节破坏， 来源于类风湿性滑膜细胞的胶原酶已经 在这一过程中受到牵连。 中性粒细胞是一种循环细胞， 在炎症部位，包括类风湿性滑膜细胞， 在这一过程中受到牵连 中性粒细胞是一种循环系统， 在炎症部位（包括类风湿性关节炎）积聚的细胞 滑液 该细胞具有至少三种溶胶原蛋白的 酶：一种降解I型、II型和III型的间质胶原酶 降解III型和IV型胶原的弹性蛋白酶;和 明胶酶降解天然V型胶原蛋白以及变性 胶原蛋白 这些酶在类风湿性关节炎中的相对作用 过程未知。 该提案的第一个目标是净化和 间质胶原酶和明胶酶的表征 人嗜中性粒细胞。 这将使用标准色谱法 技术与单克隆抗体的额外援助。 活性 这些酶的潜在形式将以特定的 关注激活过程中发生的变化。 一组单克隆抗体将针对这些酶产生， 以下目的：1）作为纯化中的助剂， 酶的表征; 2）确定 某些酶功能，如底物结合和实际底物 切割;和3）作为特异性探针用于检测 滑液 后一个目标将包括发展 特异性免疫测定，其将特异性地识别胶原酶 来源于中性粒细胞。 这些测定将用于定量这些 类风湿关节液中的胶原酶。 类风湿滑液中存在的物质能够 激活这些酶的潜在形式将被纯化 标准色谱技术。 这些物质将 其特征在于它们的分子结构和蛋白水解 活动