STRUCTURE AND BIOLOGICAL FUNCTION OF THE MONOKINE ECEF

单因子 ECEF 的结构和生物学功能

• 批准号: 3455273
• 负责人: DAVID S SIBERSTEIN
• 金额: $ 11.76万
• 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/3455273
• 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， 组织和杀死寄生生物。 嗜酸性粒细胞的其他产物 增加微血管渗透性并募集炎性细胞， 组织部位，从而对患者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， 调节嗜酸性粒细胞功能对于 免疫学和医学。 早期的研究表明，LPS刺激的单核细胞或PMA/LPS- 刺激U937细胞产生一种因子， 体外细胞毒功能。 该因子（ECEF）现已纯化。 部分氨基酸序列的测定表明，ECEF是一个 新型单核细胞 在家兔体内制备了ECEF特异性抗体。 拟议的研究涉及结构和生物学功能 的ECEF。 编码ECEF的cDNA的分子克隆计划在 为了确定ECEF的一级结构， 用于功能研究的基因定义的ECEF的量。 cDNA和 将使用单克隆抗体探针和肽图谱进行比较 天然ECEF的结构转化为重组ECEF。 的条件 将研究ECEF诱导单核细胞合成和释放。 的 外周血单核细胞和组织单核细胞产生ECEF 将分析小鼠和人类受试者的嗜酸性肉芽肿 血吸虫病 ECEF对嗜酸性粒细胞的作用机制 体外将进行研究，特别强调的阈值 激活刺激和分泌速率。 ECEF对其他人的影响 将使用功能性和 受体结合测定。