MOLECULAR BIOLOGY OF SCHISTOSOME-SNAIL INTERACTIONS

血吸虫-蜗牛相互作用的分子生物学

• 批准号: 3136005
• 负责人: WILLARD O GRANATH
• 金额: $ 15.62万
• 依托单位: UNIVERSITY OF MONTANA
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-07-01 至 1995-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3136005
• 关键词: Biomphalaria; SDS polyacrylamide gel electrophoresis; Schistosoma; Schistosoma mansoni; antibody; binding proteins; blood cells; blood proteins; cell biology; disease vectors; fluorescence microscopy; hemolymph; high performance liquid chromatography; host organism interaction; immunofluorescence technique; immunoregulation; laboratory mouse; laboratory rabbit; molecular biology; monoclonal antibody; peptides; protein structure function; radiotracer; scanning electron microscopy; schistosomiasis; surface antigens

The overall objective of this research is to elucidate the molecular, biochemical and cellular events responsible for determining susceptibility or resistance of the snail, Biomphalaria glabrata, to infection with the human blood fluke, Schistosoma mansoni. The system used consists of the NIH-Sm-PR-1 strain of S. mansoni, to which PR albino, M-line B. glabrata are susceptible, and the pigmented 10-R2 and the albino 13-16-R1 strains which are actively resistant to the parasite. There are three major goals of this proposal: 1. During the past grant period a 55 kDa polypeptide in the serum of 10-R2 B. glabrata that binds to S. mansoni sporocysts was discovered. Interestingly, similar molecular weight proteins from M-line serum did not bind to the parasite. Therefore, the structure and function of serum polypeptides (from the three snail strains) will be characterized using electrophoretic and peptide mapping techniques. Antibodies to these polypeptides will be generated, and immunofluorescent antibody tests (IFAT) and immunoscanning electron microscopy (ISEM) will be used to assess their binding properties. Lastly, these reagents will be used in important functional analysis experiments using several in vitro and in vivo systems. 2. During the past grant period an 80 kDa surface-polypeptide unique to hemocytes from resistant B. glabrata was identified. Such a polypeptide may play a crucial role in mediating larval schistosome destruction. Therefore, hemocytes will be thoroughly characterized by surface-labeling, electrophoretic and peptide mapping techniques. Next, antibodies to the major surface proteins will be generated, and IFAT and ISEM will be used to determine their binding characteristics. Furthermore, these reagents will be used in a variety of important functional analysis experiments. 3. Since the mechanism by which hemocytes respond to schistosome infection is unknown, hemocyte formation in B. glabrata will be studied by following the fate of injected thymidine-14C. Hemocyte activation will be examined by measuring RNA and protein synthesis using radioactive precursors. Finally, surface characteristics of activated hemocytes, and the functional relevance of these cells will be evaluated using several in vitro systems. This research will make a significant contribution to the understanding of how vector competence is manifested in schistosome-snail interactions, which will be useful in designing rational vector control programs.

这项研究的总体目的是阐明分子， 负责确定易感性的生化和细胞事件 或蜗牛的抗性，生物掌la glabrata，以感染 人类血液氟，曼森血吸虫。 使用的系统由 NIH-SM-PR-1链球菌菌株，pr blino，m-line B. glabrata 易感，色素为10-R2和白化病13-16-R1菌株 对寄生虫具有积极抗性。 有三个主要目标 该提议： 1。在过去的赠款期间，在10-R2的血清中有55 kDa多肽 B.与曼氏链球菌孢子囊肿结合的glabrata被发现。 有趣的是，来自M系血清的类似分子量蛋白没有 与寄生虫结合。 因此，血清的结构和功能 多肽（来自三个蜗牛菌株）将使用 电泳和肽映射技术。 这些抗体 将生成多肽，并免疫荧光抗体测试（IFAT） 和免疫统计电子显微镜（ISEM）将用于评估其 结合特性。 最后，这些试剂将用于重要 使用几种体外和体内系统的功能分析实验。 2。在过去的授予期间 鉴定出来自抗性的B. glabrata的血细胞。 这样的多肽 可能在介导幼虫斑点破坏中起着至关重要的作用。 因此，血细胞将以表面标记为彻底的特征， 电泳和肽映射技术。 接下来，抗体 将生成主要的表面蛋白质，IFAT和ISEM将用于 确定其结合特征。 此外，这些试剂将 用于各种重要的功能分析实验。 3。由于血细胞应对血吸虫感染的机制 尚不清楚，将通过以下 注射胸苷-14c的命运。 将检查血细胞激活 通过使用放射性前体测量RNA和蛋白质合成。 最后，活化血细胞的表面特征和功能 这些细胞的相关性将使用多种体外系统评估。 这项研究将为理解做出重大贡献 矢量能力如何体现在Scistosom-Snail相互作用中， 这将在设计理性矢量控制程序中有用。