MOLECULAR ANALYSIS OF APICAL ORGANELLES OF PLASMODIUM

疟原虫顶端细胞器的分子分析

• 批准号: 2442531
• 负责人: John H Adams
• 金额: $ 11.33万
• 依托单位: UNIVERSITY OF NOTRE DAME
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-07-01 至 1999-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2442531
• 关键词: Plasmodium; SDS polyacrylamide gel electrophoresis; antigen antibody reaction; apical membrane; binding proteins; blood chemistry; enzyme linked immunosorbent assay; erythrocyte membrane; gene expression; genetic polymorphism; human subject; intracellular parasitism; laboratory mouse; laboratory rabbit; laboratory rat; molecular cloning; nucleic acid probes; polymerase chain reaction; protein structure function; protozoal vaccine; recombinant proteins; southern blotting; species difference; Plasmodium; SDS polyacrylamide gel electrophoresis; antigen antibody reaction; apical membrane; binding proteins; blood chemistry; enzyme linked immunosorbent assay; erythrocyte membrane; gene expression; genetic polymorphism; human subject; intracellular parasitism; laboratory mouse; laboratory rabbit; laboratory rat; molecular cloning; nucleic acid probes; polymerase chain reaction; protein structure function; protozoal vaccine; recombinant proteins; southern blotting; species difference

Plasmodium merozoite actively enter host erythrocytes by a complex but poorly understood invasion process. Molecules from the micronemes, an organelle of the merozoite apical complex, appear to facilitate parasite invasion by binding to specific receptors on the erythrocyte's surface. Different Plasmodium species use different erythrocyte receptors, but these parasite molecules which recognize the different receptors are genetically related. Members of this family of erythrocyte binding proteins, herein termed merozoite Microneme Protein-1 (MP-1), include the P. vivax and P. knowlesi Duffy binding protein family and the P. falciparum sialic acid binding protein (EBA175). These proteins have intrinsic interest for their biological function and also because antibody to the EBP175 has been shown to inhibit parasite development. Identification of conserved and immunogenic regions of these proteins which induce an inhibitory immune response will be important for the development of an asexual stage malaria vaccine. Genetic analysis of the MP-1 gene family is necessary to identify conserved and polymorphic regions. Towards this goal I have examined laboratory strains of P. knowlesi and field isolates of P. vivax. I have identified polymorphisms in the MP-1 genes of both species. I propose to further characterize polymorphism in field isolates of P. vivax in order to determine if the vivax MP-1 has more than one allele and to identify conserved and polymorphic regions of the MP-1. Laboratory models for the study of the MP-1 family will be established using the rodent malaria parasites P. berghei and P. yoelii. The MP-1 genes of P. berghei and P. yoelii have been identified in my laboratory and will be cloned and characterized. The MP-1 genes of P. vivax, P. berghei and P. voelii will be expressed as recombinant proteins and used to assay immune reactions to MP-1 in their respective hosts. Full length and separate regions of the MP-1 will be expressed as recombinant proteins and tested for growth inhibition in vaccinated animals. Monoclonal antibodies will be produced against the P. berhei MP-1 for use in characterizing immunogenic epitopes and functional regions. The experiments outlined in this proposal will help identify and characterize the essential functional regions of these important erythrocyte binding proteins so they can be developed as effective vaccines.

疟原虫通过复合物积极进入宿主红细胞 了解入侵过程还差。 来自微分子的分子，一个 Merozoite顶端综合体的细胞器似乎有助于寄生虫 通过与红细胞表面上的特定受体结合来侵袭。 不同的疟原虫使用不同的红细胞受体，但 这些识别不同受体的寄生虫分子是 遗传相关。 这个红细胞结合的家族的成员 蛋白质（以下称为梅罗唑岩微生物蛋白-1（MP-1））包括 P. vivax和P. knowlesi duffy结合蛋白家族和P. 恶性唾液酸结合蛋白（EBA175）。 这些蛋白质具有 其生物学功能的内在兴趣，也是因为 对EBP175的抗体已显示可抑制寄生虫的发育。 鉴定这些蛋白质的保守和免疫原状区域 诱导抑制免疫反应对 开发无性阶段疟疾疫苗。 MP-1基因家族的遗传分析对于鉴定是必要的 保守和多态区域。 对于这个目标，我已经检查了 P. knowlesi的实验室菌株和Vivax的田间分离株。 我有 在这两种物种的MP-1基因中鉴定出多态性。 我建议 为了进一步表征假霉菌的田间分离株中的多态性 为了确定Vivax MP-1是否具有多个等位基因和 确定MP-1的保守和多态区域。 实验室 使用MP-1家族的研究模型将使用 啮齿动物疟疾寄生虫P. Berghei和P. Yoelii。 P.的MP-1基因 Berghei和P. Yoelii已在我的实验室中确定，将是 克隆和特征。 Vivax，P。Berghei和P.的MP-1基因 VOELII将表示为重组蛋白，用于测定免疫 在其各自宿主中对MP-1的反应。 全长，分开 MP-1的区域将表示为重组蛋白并测试 用于疫苗接种动物的生长抑制作用。 单克隆抗体会 使用P. berhei MP-1生产以用于表征 免疫原性表位和功能区域。 实验概述了 在此提案中，将有助于识别和表征必不可少的 这些重要的红细胞结合蛋白的功能区域 它们可以作为有效的疫苗开发。