STRUCTURE AND BIOLOGICAL FUNCTION OF THE MONOKINE ECEF

单因子 ECEF 的结构和生物学功能

• 批准号: 3455272
• 负责人: DAVID S SIBERSTEIN
• 金额: $ 9.37万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-08-01 至 1994-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3455272
• 关键词: antibody specificity; autoradiography; bioassay; cell mediated cytotoxicity; chemical structure function; complementary DNA; eosinophil; eosinophilia; genetic manipulation; high performance liquid chromatography; human subject; immunological substance; inflammation; laboratory mouse; laboratory rabbit; leukocyte activation /transformation; molecular cloning; monokines; nucleic acid probes; pathologic process; protein sequence; radioimmunoassay; tissue /cell culture

Activated eosinophils release toxic cationic proteins that can damage tissues and kill parasitic organisms. Other products of eosinophils increase microvascular permeability and recruit inflammatory cells to tissue sites, thus having a major impact on the pathophysiology of eosinophilia-associated diseases. For this reason, the factors that regulate eosinophil function are of considerable importance to immunology and medicine. Earlier studies showed that LPS-stimulated monocytes or PMA/LPS- stimulated U937 cells produce a factor that enhances eosinophil cytotoxic function in vitro. This factor (ECEF) has now been purified. A partial amino acid sequence determination suggests that ECEF is a novel monokine. ECEF-specific antibody has been produced in rabbits. The proposed research deals with the structure and biological functions of ECEF. The molecular cloning of the cDNA encoding ECEF is planned in order to determine the primary structure of ECEF and to produce quantities of genetically-defined ECEF for functional studies. cDNA and monoclonal antibody probes and peptide mapping will be used to compare the structure of natural ECEF to recombinant ECEF. Conditions that induce monocyte synthesis and release of ECEF will be studied. The production of ECEF by peripheral blood monocytes and tissue monocytes in eosinophilic granulomas will be analyzed for mice and human subjects with schistosomiasis. The mechanism of action of ECEF on eosinophils in vitro will be studied, with particular emphasis on the threshold of activation stimuli and rates of secretion. The effects of ECEF on other types of inflammatory cells will be evaluated, using functional and receptor binding assays.

活化的嗜酸性粒细胞释放有毒的阳离子蛋白，可破坏 组织和杀死寄生生物。嗜酸性粒细胞的其他产物 增加微血管通透性，募集炎性细胞 组织部位，从而对心脏的病理生理学产生重大影响。 嗜酸性粒细胞增多症相关疾病。因此，这些因素 调节嗜酸性粒细胞功能对 免疫学和医学。 早期的研究表明，内毒素刺激单核细胞或PMA/内毒素- 受刺激的U937细胞产生一种增强嗜酸性粒细胞的因子 体外细胞毒作用。该因子(ECEF)现已提纯。 部分氨基酸序列测定表明，ECEF是一种 新奇的单倍体。已经在兔体内产生了ECEF特异性抗体。 拟议的研究涉及结构和生物学功能。 ECEF的。编码ECEF的cDNA的分子克隆计划在 以确定ECEF的一级结构并生产 用于功能研究的大量遗传定义的ECEF。CDNAs和 将使用单抗探针和多肽图谱来比较 将天然ECEF的结构转化为重组ECEF。符合以下条件的条件 诱导单核细胞合成和释放ECEF将进行研究。这个 外周血单核细胞和组织单核细胞产生ECEF的研究 将分析小鼠和人类受试者的嗜酸性肉芽肿 患有血吸虫病。ECEF对小鼠嗜酸粒细胞的作用机制 将对体外培养进行研究，特别强调 激活刺激和分泌率。ECEF对其他组织的影响 炎症细胞的类型将被评估，使用功能性和 受体结合分析。