STRUCTURE AND FUNCTION OF HUMAN SECRETORY COMPONENT

人体分泌成分的结构和功能

• 批准号: 3131510
• 负责人: Randall M Goldblum
• 金额: $ 9.93万
• 依托单位: UNIVERSITY OF TEXAS MED BR GALVESTON
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-07-01 至 1988-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3131510
• 关键词: affinity chromatography; chemical structure function; conformation; enzyme linked immunosorbent assay; epithelium; gel electrophoresis; high performance liquid chromatography; immunofluorescence technique; immunoglobulin A; monoclonal antibody; protease inhibitor; proteolysis; receptor mediated endocytosis; secretion; secretory immune system

Secretory component (SC) plays a central role in the mucosal immune system. It functions as an intergal membrane receptor, transcellular transporter, and protease inhibitor for its ligand polymeric IgA. A panel of monoclonal antibodies to human SC, characterized in our laboratories, offers a unique opportunity to study the immunobiology of SC. Our basic objective is to examine the structural characteristics which allow SC to function in its multiple specialized roles. Clinical studies will seek evidence for heterogeneity in size and structure of SC in different secretions and from different individuals. These studies may provide evidence for a modulatory role for SC and insight into alterations in SC metabolism in malignant epithelial cells. The SC from various secretions will be isolated by immunoaffinity chromatography and certain preparations will be modified by deglycosylation and limited and enzymatic proteolysis. Initially SDS polyacrylamide gel electrophoresis, HPLC and reaction with monoclonal antibodies will be used to characterize the size and epitopic structure of the products. Subsequently, amino acid and carbohydrate composition and amino acid sequence analysis will be determined for certain SC fragments. The in vitro function of the SC preparations will be assessed by evaluating polymeric IgA binding and inhibition of IgA proteolysis. Intracellular events will be examined using a colon carcinoma derived cell line. The intracellular pathway of SC will be followed by immunocytochemistry using monoclonal antibodies which distinguish the various physical forms of SC. Glycosylation will be inhibited with tunicamycin and attention focused on the role of carbohydrate in SC cellular function. Human IgG antibodies to SC will be used to probe the in vivo conformation of the receptor form of SC by comparing the spectrum of antibodies in the serum and secretions of individuals. A modulatory role for these antibodies will be tested in an in vitro model of SC function. Deliniation of the normal structure and function of SC will enhance our understanding of mucosal immunity and its control. Therapeutic applications of this knowledge may include the development of methods for targeting active or passive immunity or other therapeutic agents to the site of mucosal infections. The proposed studies may also elucidate alterations in SC metabolism in malignant epithelial cells which may allow direction of cytotoxic or chemotherapeutic agent selectively to these tumors.

分泌成分（Secretory component，SC）在粘膜免疫中起着重要作用， 系统 它作为一种跨细胞膜受体， 转运蛋白和其配体多聚伊加的蛋白酶抑制剂。 面板 在我们的实验室中鉴定的抗人SC的单克隆抗体， 提供了一个独特的机会，研究SC的免疫生物学。我们的基本 目的是检查允许SC的结构特征， 发挥其多个专业作用。 临床研究将寻求 不同地区SC大小和结构的异质性证据 分泌物和不同的人。 这些研究可以提供 SC调节作用的证据和对SC改变的认识 恶性上皮细胞的代谢。 将通过免疫亲和法从各种分泌物中分离SC 某些制剂将通过去糖基化进行修饰 和有限的酶促蛋白水解。 SDS聚丙烯酰胺凝胶 将使用电泳、HPLC和与单克隆抗体的反应 以表征产物的尺寸和外延结构。 随后，氨基酸和碳水化合物的组成和氨基酸 将确定某些SC片段的序列分析。 的in 通过评价SC制剂的体外功能， 多聚伊加结合和伊加蛋白水解的抑制。 细胞内 将使用结肠癌衍生的细胞系检查事件。 的 SC的细胞内途径将通过免疫细胞化学进行， 单克隆抗体，区分SC的各种物理形式。 衣霉素可抑制糖基化， 碳水化合物在SC细胞功能中的作用 SC的人IgG抗体将用于探测体内构象 SC受体形式通过比较抗体的光谱， 个体的血清和分泌物。 这些调节作用 将在SC功能的体外模型中测试抗体。 阐明SC的正常结构和功能，将有助于我们更好地了解SC的功能， 了解粘膜免疫及其控制。 治疗 这些知识的应用可能包括开发方法， 将主动或被动免疫或其它治疗剂靶向于 粘膜感染部位。 拟议的研究还可以阐明 恶性上皮细胞中SC代谢的改变， 选择性地将细胞毒性剂或化学治疗剂导向这些 肿瘤的