CHARACTERIZATION OF HUMAN MALARIA CRISIS-FORM FACTOR

人类疟疾危机形成因素的特征

• 批准号: 3134576
• 负责人: JAMES B JENSEN
• 金额: $ 8.96万
• 依托单位: MICHIGAN STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-09-30 至 1989-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3134576
• 关键词: Plasmodium falciparum; antibody; antiserum; cellular immunity; cytokine; high performance liquid chromatography; human tissue; interferons; interleukin 2; ion exchange chromatography; lymphokines; malaria; microorganism culture; microorganism immunology; monoclonal antibody; monokines; protozoal antigen; tissue /cell culture; tropical medicine; tumor necrosis factor alpha

Despite the evidence that functional malarial immunity is multifactorial, involving complex interactions of humoral and cellular immune responses, present knowledge of efferent mechanisms of cell-mediated immunity is limited. However, immunologic resolution of murine and primate malarias are often associated with intraerythrocytic retardation of parasite development resulting in moribund parasites known as crisis-forms. Human malaria immune serum from Sudan retards Plasmodium falciparum in vitro by a non-antibody substance termed crisis-form factor, CFF. Epidemologic evidence reveals that CFF is closely associated with clinical malaria immunity in Sudan, where serum CFF concentrations increase with malaria endemicity, and fluctuate with seasonal malaria transmission. Thus, CFF may be part of the acquired immunity to malaria, and a product of cell-mediated responses. It is the specific aim of this project to purify and characterize CFF; to determine its source and the stimulatory mechanism, or antigen associated with its production, and to determine how it retards parasite development. Purification of CFF from human immune serum will be conducted using standard column chromatographic procedures followed by high-performance liquid chromatography using FPLC. Polyclonal and monoclonal antibodies will be developed against purified CFF to be used for ultimate purification by affinity chromatography and for analysis of the cellular source and stimulatory cascade leading to CFF production. The relationship between CFF and known cytokines will be studied by comparing effects of huIFNGamma, Il 1,2,3, TNF and other known lymphokines and monokines on cultures of P. falciparum. Antiplasmodium activity of secretory products of human mononuclear cells stimulated in vitro with known activators will be compared to activity of CFF-containing human immune sera to determine the cell type responsible for CFF production, and as a prelude to investigation of the in vivo production of CFF and the possible role of malarial antigens in its production. The mode of action of CFF will be studied by monitoring incorporation of radiolabelled precursors of parasite protein and nucleic acid synthesis, glucose utilization, mitochondrial activity and other known parameters of parasite metabolism in highly synchronized cultures of P. falciparum.

尽管有证据表明功能性疟疾免疫是多因素的， 涉及体液和细胞免疫应答的复杂相互作用， 目前对细胞介导免疫的传出机制的认识是 有限公司 然而，小鼠和灵长类疟疾的免疫学解决方案 通常与红细胞内寄生虫迟滞有关 发展导致垂死的寄生虫称为危机形式。 人类 苏丹疟疾免疫血清对恶性疟原虫的体外抑制作用 非抗体物质称为危机形成因子CFF。 流行病学的 有证据表明，CFF与临床疟疾密切相关 苏丹的免疫力，那里的血清CFF浓度随着疟疾而增加 疟疾的流行率随着季节性疟疾传播而波动。 因此，CFF 可能是获得性免疫疟疾的一部分， 细胞介导的反应 这个项目的具体目标是净化 并表征CFF;以确定其来源和刺激性 机制，或与其生产相关的抗原，并确定如何 它能延缓寄生虫的发育。 人免疫细胞CFF的纯化 将使用标准柱色谱法进行血清分析 然后使用FPLC进行高效液相色谱分析。 多克隆 将开发针对纯化CFF的单克隆抗体， 用于通过亲和色谱法进行最终纯化和用于分析 导致CFF产生的细胞来源和刺激级联。 的 CFF与已知细胞因子之间的关系将通过比较来研究 IFN γ、IL 1、2、3、TNF和其他已知的淋巴因子的作用， 恶性疟原虫培养物上的单核因子。 抗疟原虫活性 体外刺激人单核细胞的分泌产物 将已知的活化剂与含CFF的人 免疫血清以确定负责CFF产生的细胞类型，和 作为研究CFF的体内产生和 疟疾抗原在其生产中的可能作用。 作用方式 将通过监测放射性标记的 寄生虫蛋白质和核酸合成的前体，葡萄糖 利用率、线粒体活性和寄生虫的其他已知参数 恶性疟原虫高度同步培养物中的代谢。