HUMORAL FACTORS IN INFLAMMATION

炎症中的体液因素

• 批准号: 3338174
• 负责人: TONY E HUGLI
• 金额: $ 21.51万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-05-08 至 1994-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3338174
• 关键词: aminoacid analyzer; anaphylatoxins; antibody formation; cell type; cellular immunity; chemical structure function; complement; complement receptor; crosslink; guinea pigs; high performance liquid chromatography; humoral immunity; inflammation; iodine; laboratory mouse; laboratory rabbit; laboratory rat; macrophage; monocyte; protein biosynthesis; radionuclides; radiotracer; receptor binding; receptor expression; tissue /cell culture

The anaphylatoxins mediate spasmogenic, chemotactic and immunoregulatory responses primarily through specific receptor interactions with granulocytes and monocytes. The primary targets of the ligand C5a in man are believed to be the neutrophil and the monocyte (macrophage). Involvement of other cell types such as platelets, lymphocytes, endothelial cells and mast cells may occur as an indirect consequence of granulocyte or monocyte activation and secondary mediator release. Evidence exists that a C5a receptor of approximately 40,000 kDa occurs on the human neutrophil surface and that this membrane component has an affinity for the ligand of 1-3 nM Kd. The human neutrophil has no receptor for the C3a anaphylatoxin. This proposal concerns both C5a and C3a receptors on monocytes (macrophages). Preliminary evidence indicates that monocytes and tissue macrophages from human and animal sources have C3a and C5a receptors. Using chemical cross-linking techniques to visualize these membrane components, a C3a binding protein of approximately 1000,000 kDa and a C5a binding protein of approximately 40,000 kDa were observed. We plan to isolate these receptors, characterize them in terms of carbohydrate content and solubility. Monospecific immune reagents will be prepared to study functional attributes of the C5a receptor on macrophages and for use in purification of the receptor in quantities adequate for partial sequence analysis. Probes designed from protein sequence analysis will be used for complete molecular analysis of the membrane component. We have observed that transformed murine macrophage cell lines express C5a receptors. When these cell lines are injected intraperitoneally into mice, tumors are grown that contain large quantities of the macrophages having markedly enhanced number of C5a receptors (i.e. upregulated from membrane components desired). We plan to use this discovery to advantage in isolation and characterization of the macrophage chemotactic (C5a) receptor. A biologic, biochemical and molecular analysis of the anaphylatoxic receptors on monocytes and macrophages should lead to a better understanding of the role that humoral factors play in modulating immune and inflammatory responses. Circulating monocytes and tissue macrophages appear to play a pivotal role in host defense mechanisms. Early events in target cell activation are elicited by factors such as anaphylatoxins and bacterial products indicating that they represent activators of major biologic consequence.

过敏毒素介导痉挛性、趋化性和 主要通过特定受体的免疫调节反应 与粒细胞和单核细胞的相互作用。 主要目标 据信，人类配体 C5a 是中性粒细胞， 单核细胞（巨噬细胞）。 其他细胞类型的参与，例如 血小板、淋巴细胞、内皮细胞和肥大细胞可能发生 作为粒细胞或单核细胞激活的间接结果 和次级介体释放。 有证据表明 C5a 人类中性粒细胞上存在约 40,000 kDa 的受体 表面并且该膜成分对 1-3 nM Kd 的配体。 人类中性粒细胞没有受体 C3a过敏毒素。 该提案涉及单核细胞上的 C5a 和 C3a 受体 （巨噬细胞）。 初步证据表明单核细胞和 人类和动物来源的组织巨噬细胞含有 C3a 和 C5a 受体。 使用化学交联技术可视化 这些膜成分，大约是 C3a 结合蛋白 1000,000 kDa 和约 40,000 kDa 的 C5a 结合蛋白 被观察到。 我们计划分离这些受体，表征 它们的碳水化合物含量和溶解度。 单特异性 将制备免疫试剂来研究功能属性 巨噬细胞上的 C5a 受体并用于纯化 受体的数量足以进行部分序列分析。 根据蛋白质序列分析设计的探针将用于 膜成分的完整分子分析。 我们有 观察到转化的小鼠巨噬细胞系表达 C5a 受体。 当这些细胞系被腹腔注射时 小鼠体内长出的肿瘤含有大量的 巨噬细胞的 C5a 受体数量显着增加（即 从所需的膜成分上调）。 我们计划使用这个 发现有利于分离和表征 巨噬细胞趋化 (C5a) 受体。 生物、生化 单核细胞过敏毒性受体的分子分析 和巨噬细胞应该可以更好地理解其作用 体液因子在调节免疫和炎症中发挥作用 回应。 循环单核细胞和组织巨噬细胞似乎 在宿主防御机制中发挥着关键作用。 早期事件发生在 靶细胞激活是由以下因素引起的 过敏毒素和细菌产物表明它们 代表主要生物学后果的激活剂。