CHARACTERIZATION OF HUMAN MALARIA CRISIS-FORM FACTOR

人类疟疾危机形成因素的特征

• 批准号: 3564417
• 负责人: JAMES B JENSEN
• 金额: $ 11.67万
• 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/3564417
• 关键词: Plasmodium falciparum; antibody; antiserum; cellular immunity; cytokine; high performance liquid chromatography; human tissue; interferons; interleukin 1; interleukin 2; ion exchange chromatography; laboratory mouse; laboratory rabbit; 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.

尽管有证据表明疟疾的功能性免疫是多因素的， 涉及体液和细胞免疫反应的复杂相互作用， 目前关于细胞免疫传出机制的知识是 有限的。然而，小鼠和灵长类疟疾的免疫分辨 常与寄生虫的红细胞内滞留有关 发展导致濒临死亡的寄生虫，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 称为危象因子(CFF)的非抗体物质。流行病学 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浓度随着疟疾的增加而增加 地方性，并随季节性疟疾传播而波动。因此，CFF 可能是对疟疾后天免疫力的一部分，也是 细胞介导的反应。提纯是本项目的具体目标 并对CFF进行表征，以确定其来源和刺激作用 机制，或与其产生相关的抗原，并确定如何 它会阻碍寄生虫的发展。人免疫来源CFF的纯化 血清将使用标准柱层析程序进行处理 然后用FPLC进行高效液相色谱分离。多克隆 并将开发针对纯化的CFF的单抗以供使用 用于亲和层析的最终纯化和用于分析 导致CFF产生的细胞源和刺激级联。这个 将通过比较来研究CFF与已知细胞因子的关系 环磷酰胺、IL-1、2、3、肿瘤坏死因子等淋巴因子的作用及机制 单因子对恶性疟原虫培养的影响。抗疟原虫活性的研究 人单核细胞体外刺激的分泌产物 已知的激活剂将与含有CFF的人的活性进行比较 免疫血清以确定负责CFF产生的细胞类型，以及 作为研究CFF体内生产的前奏和 疟疾抗原在其生产中的可能作用。行动模式 将通过监测放射性标记的掺入来研究CFF 寄生虫蛋白质和核酸合成的前体，葡萄糖 寄生虫的利用、线粒体活性和其他已知参数 高度同步化培养的恶性疟原虫的代谢。