BIOLOGICAL STUDIES OF HUMAN INTERLEUKIN I

人白细胞介素 I 的生物学研究

• 批准号: 3176051
• 负责人: Lawrence B Lachman
• 金额: $ 6.14万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 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 1987-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3176051
• 关键词: acute phase protein; affinity chromatography; antiserum; cell growth regulation; collagenase; enzyme linked immunosorbent assay; gel electrophoresis; high performance liquid chromatography; human tissue; immunological substance; inflammation; leukemia; leukocyte activation /transformation; leukopoiesis; macrophage; monoclonal antibody; monocyte; neoplastic cell; pyrogens

The role of monocytes and macrophages in inflammatory and immune responses has yet to be characterized at the molecular level. Although it is clear that activated macrophages an monocytes release a large number of biological mediators at the site of inflammation, the effect of these mediators upon lymphocytes, fibroblasts, endothelial and possibly other cells remains to be determined. A single pluripotent monokine, Interleukin 1 (IL 1), has been demonstrated to affect the inflammatory process at several stages. Although Interleukin 1 has been extensively investigated, it has not been purified to homogeneity in a biologically active form. In addition, monoclonal antibodies to IL 1, which are required to measure IL 1 in biological fluids, to affinity purify IL 1, and to isolate polysomal mRNA for gene cloning, have yet to be developed. Once the primary sequence of IL 1 has been determined, it will be possible to predict antigenic regions to select possible sequences for nuceotide probes specific for IL 1 coding m RNA, and to indicate possible amino acid sequences for antagonists of IL 1 which may be potential anti-inflammatory and anti arthritic agents. It is proposed to determine the primary structure of human IL 1 purified to homogeneity by SDS-polyacrylamide gel electrophoresis. (2) To prepare homogeneously purified human IL 1 which retains biological activity. (3) To prepare monoclonal antibodies and heteranisera to human IL 1. (4) To use the abovementioned antibodies to develop ELISA assays which can quantitate IL 1 levels in complex bodily fluids, and to prepare affinity reagents which can remove IL 1 from these fluids. (5) To continue biological studies using homogeneously purified, biologically active IL 1 to unequivocally establish that a single molecule elevates fever, induces fibroblast proliferation stimulates collagenase release from fibroblasts, stimulates acute phase reactants when injected in vivo, induces B and T lymphocyte proliferation and granulopoeisis, and possibly affects vascular permeability.

单核细胞和巨噬细胞在炎症和免疫反应中的作用 还有待于在分子水平上进行鉴定。 虽然很明显 激活的巨噬细胞和单核细胞释放大量的 炎症部位的生物介质，这些介质的作用 介质对淋巴细胞，成纤维细胞，内皮细胞和可能的其他 细胞仍有待确定。 一种单一的多能性单核因子，白细胞介素 1（IL 1），已被证明可以影响炎症过程， 几个阶段。 虽然白细胞介素1已被广泛研究， 它还没有被纯化成生物活性形式的均一性。 在 此外，IL 1的单克隆抗体，这是测量IL 1所必需的 在生物流体中，亲和纯化IL 1，并分离多聚体 用于基因克隆的mRNA还有待开发。 一旦主序列 IL 1的含量已经确定，可以预测抗原性 选择IL 1特异性核苷酸探针的可能序列的区域 编码mRNA，并指示拮抗剂的可能氨基酸序列 IL-1可能是潜在的抗炎和抗关节炎 剂. 本研究旨在确定人IL-1的一级结构 通过SDS-聚丙烯酰胺凝胶电泳纯化至均一。 (2)到 制备均匀纯化的人IL-1， 活动 (3)目的制备抗人源性大肠杆菌单克隆抗体和异源性大肠杆菌单克隆抗体 IL 1. (4)使用上述抗体开发ELISA测定法 它可以定量复杂体液中的IL 1水平， 可以从这些液体中去除IL 1的亲和试剂。 (5)继续 使用均匀纯化的生物活性IL 1的生物学研究 明确地确定一个单一的分子会引起发烧， 成纤维细胞增殖刺激胶原酶从成纤维细胞释放， 在体内注射时刺激急性期反应物，诱导B和T 淋巴细胞增殖和粒细胞生成，并可能影响血管 磁导率