IMMUNOBIOLOGY OF SCHISTOSOME-MOLLUSK INTERACTIONS

血吸虫-软体动物相互作用的免疫生物学

• 批准号: 3126215
• 负责人: TIMOTHY P. YOSHINO
• 金额: $ 15.26万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-07-01 至 1997-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3126215
• 关键词: Biomphalaria; SDS polyacrylamide gel electrophoresis; Schistosoma mansoni; affinity chromatography; alternatives to animals in research; cellular immunity; cytotoxicity; disease vectors; genetic strain; helminthic antigen; hemolymph; high performance liquid chromatography; host organism interaction; humoral immunity; immunity; immunochemistry; immunofluorescence technique; microinjections; monoclonal antibody; phagocytes; tissue /cell culture

The long-term objective of the proposed research is to define and characterize the molecular mechanisms underlying genetically-determined resistance and susceptibility in snails Biomphalaria glabrata to infection by larval Schistosoma mansoni, causative agent of human schistosomiasis mansoni. Defined susceptible (S) and resistant (R) strains of B. glabrata will be employed in comparative biochemical and immunological studies designed to provide an indepth analysis of the molecular interaction between snail hemocytes (primary anti-parasite effector cells) and the early intramolluscan stages of S. mansoni (miracidium, primary sporocyst). Surface polypeptides from S and R strain hemocytes will be labeled using a highly sensitive biotin-avidin system and subjected to SDS-PAGE analyses to compare the occurrence of individual surface polypeptides on cells of each host strain, and to determine which of the identified components serve as binding receptors for larval excretory-secretory proteins (ESP) or sporocyst tegumental surface proteins (TSP). S and R snail hemocyte cDNA expression libraries also will be constructed, and screened with hemocyte membrane-reactive antisera to identify cDNA inserts encoding potential cell surface polypeptides. Expressed recombinant hemocyte proteins will then be used in further structural comparisons between snail strains, and tested for their abilities to bind S. mansoni ESP/TSP and to alter hemocyte function as a consequence of binding. Finally, individual ESP and TSP components will be isolated by high performance liquid chromatography and preparative PAGE, and evaluated for their ability to bind S and R hemocyte surface proteins (receptors) and to modulate hemocyte-sporocyst interactions. this focused research plan directly addresses questions related to the biochemical basis for hemocyte recognition and/or activation by early schistosome larvae. It is anticipated that the information generated by this research will provide important insights into the basic mechanisms of pathogen recognition by the molluscan internal defense system, and eventually may be applied to novel approaches to control invertebrate vectors of human disease.

拟议研究的长期目标是定义和 表征遗传确定的分子机制 蜗牛的耐药性和易感性 幼虫血吸虫Mansoni的感染，人类的病因药物 血吸虫病Mansoni。 定义的易感性和抗性（R） B. glabrata菌株将用于比较生化和 免疫研究旨在提供对 蜗牛血细胞之间的分子相互作用（主要抗寄生虫 效应细胞）和曼氏链球菌的早期内膜内阶段 （Miracidium，原发性孢子囊）。 S和R的表面多肽 菌株血细胞将使用高度敏感的生物素 - 阿维丁标记 系统并进行SDS-PAGE分析，以比较 每个宿主应变细胞上的单个表面多肽，然后 确定哪些确定的成分用作结合受体 用于幼虫排泄分泌蛋白（ESP）或孢子囊 表面蛋白（TSP）。 S和R蜗牛血细胞cDNA表达库 还将构建并用血细胞膜反应筛选 抗血清以识别cDNA插入编码电势细胞表面的插入 多肽。 然后将使用表达的重组血细胞蛋白 在蜗牛菌株之间的进一步结构比较中，并测试了 它们结合链球菌ESP/TSP并改变血细胞功能的能力 由于结合的结果。 最后，个人ESP和TSP组件 将通过高性能液相色谱和 准备页面，并评估其绑定S和R的能力 血细胞表面蛋白（受体）并调节血细胞 - 孢子囊肿 互动。 这个重点研究计划直接解决了问题 与血细胞识别和/或的生化基础有关 早期血块幼虫的激活。 预计 这项研究产生的信息将提供重要的见解 进入软体动物的病原体识别的基本机制 内部防御系统，最终可能应用于新颖 控制人类疾病的无脊椎动物载体的方法。