New Opportunistic Infections in AIDS Microsporidiosis

艾滋病微孢子虫病的新机会性感染

• 批准号: 7406048
• 负责人: Louis M. Weiss
• 金额: $ 39.92万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-04-01 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7406048
• 关键词: Acquired Immunodeficiency Syndrome; Binding; Biochemical; Cells; Cloning; Code; Complement; Cryoelectron Microscopy; Development; Disease; Encephalitozoon cuniculi; Facility Construction Funding Category; Genes; Genetic Recombination; Genome; Germination; Glutathione S-Transferase; Human; Immunocompetent; Infection; Laboratories; Lead; Location; Mass Spectrum Analysis; Microsporidia; Microsporidiosis; Molecular; Morphogenesis; Morphology; Opportunistic Infections; Organelles; Organism; Penetration; Pharmaceutical Preparations; Proteins; Proteomics; Reproduction spores; Structure; Techniques; Therapeutic Agents; Therapeutic Intervention; Tube; Yeasts; base; extracellular; insight; novel therapeutics; pathogen; three dimensional structure; tomography; transmission process; yeast two hybrid system

DESCRIPTION (provided by applicant): The microsporidia are "emerging" human and veterinary pathogens that cause disease in both immunocompromized and immunocompetent hosts. Our efforts are directed at understanding the invasion mechanism of these organisms involving the polar tube and its associated structures which form an organelle critical for cell invasion. The polar tube serves as a vehicle for transmission of infection by piercing an adjacent host cell inoculating the sporoplasm directly into that cell. The biochemical components of this structure and the mechanism of its formation during invasion remain to be definitively determined. Study of the composition, formation and function of this organelle during germination and invasion should provide a basis for the development of new strategies for control of these important parasitic protists. We plan to use a proteomics (mass spectrometry) approach to the characterization of the proteins of the polar tube (PTPs) and spore wall (SWPs) of the microsporidia. Encephalitozoon cuniculi, a human pathogenic microsporidia, will be used for these studies as the availability of its complete genome facilitates this proteomic approach. Recombination cloning will be used to express and characterize the identified proteins as this will allow high throughput expression analysis. The molecular interaction(s) of PTP1 with itself and with other PTPs will be evaluated using yeast two hybrid and GST pull- down techniques. Our characterization of these structures at the molecular level will be complemented by ultrastructural studies investigating the development and location of PTPs and SWPs during morphogenesis and eversion of the polar tube. Cryoelectron microscopy and EM tomography will provide insights into the 3D structure of the polar tube and spore wall during eversion, providing answers to fundamental questions on the organism of these structures that have not been resolved by traditional EM. For example, our observations suggest that the polar tube may be extracellular in the spore. We believe that the information gained by delineating, in detail, the function and components of the polar tube will eventually lead to novel therapeutic interventions that could limit or interdict the transmission of these emerging pathogens. RELEVANCE. These are new pathogens for which there are limited effective drugs. Studies of how these organisms cause infection should result in the development of new therapeutic agents.

描述（由申请人提供）：微孢子虫是“新兴”人类和兽医病原体，可在免疫功能低下和免疫功能正常的宿主中引起疾病​​。我们的努力旨在了解这些生物体的侵袭机制，涉及极管及其相关结构，这些结构形成对细胞侵袭至关重要的细胞器。极管作为感染传播的载体，通过刺穿相邻的宿主细胞，将孢子质直接接种到该细胞中。该结构的生化成分及其在入侵过程中形成的机制仍有待明确确定。研究这种细胞器在萌发和入侵过程中的组成、形成和功能应该为开发控制这些重要寄生原生生物的新策略提供基础。我们计划使用蛋白质组学（质谱）方法来表征微孢子虫的极管（PTP）和孢子壁（SWP）的蛋白质。 Encephalitozoon cuniculi（一种人类致病性微孢子虫）将用于这些研究，因为其完整基因组的可用性有助于这种蛋白质组学方法。重组克隆将用于表达和表征已鉴定的蛋白质，因为这将允许高通量表达分析。 PTP1 与其自身以及与其他 PTP 的分子相互作用将使用酵母二杂交和 GST Pull-down 技术进行评估。我们在分子水平上对这些结构的表征将得到超微结构研究的补充，这些研究调查了极管形态发生和外翻过程中 PTP 和 SWP 的发育和位置。冷冻电子显微镜和电磁断层扫描将深入了解外翻期间极管和孢子壁的 3D 结构，为传统电磁学尚未解决的有关这些结构的生物体的基本问题提供答案。例如，我们的观察表明极管可能位于孢子的细胞外。我们相信，通过详细描述极管的功能和成分所获得的信息最终将导致新的治疗干预措施，从而限制或阻止这些新出现的病原体的传播。关联。这些是新的病原体，有效药物有限。对这些微生物如何引起感染的研究应该会导致新治疗剂的开发。