Molecular Composition and Function of Trypanosoma cruzi Shed Vesicles

克氏锥虫脱落囊泡的分子组成和功能

• 批准号: 8147543
• 负责人: IGOR C ALMEIDA
• 金额: $ 37.68万
• 依托单位: UNIVERSITY OF TEXAS EL PASO
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-29 至 2012-05-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8147543
• 关键词: Achievement; Address; Adhesions; Affect; Affinity; Animals; Antibodies; Antigens; Binding; Binding Proteins; Biochemical; Biological; Biological Assay; Biotin; CD44 Antigens; Calcium; Cardiac; Cell Adhesion; Cell membrane; Cell surface; Cells; Chagas Disease; Chronic Phase of Disease; Complex; Confocal Microscopy; Data; Dendritic Cells; Development; Digestive System Disorders; Electrons; Event; Flow Cytometry; GPI Membrane Anchors; Galectin 3; Glycoproteins; Glycosylphosphatidylinositols; Goals; Human; Immune; Immune response; Immunity; Immunoassay; Immunoelectron Microscopy; Immunohistochemistry; In Vitro; Infection; Inflammatory; Invaded; Knock-out; Knowledge; Label; Latin America; Lead; Learning; Lectin; Liquid Chromatography; Lysosomes; Mass Spectrum Analysis; Mediating; Methods; Modification; Molecular; Mucins; Mus; Nitric Oxide; Nomads; Oligosaccharides; Parasites; Pathogenesis; Pathway interactions; Pharmaceutical Preparations; Phosphopeptides; Phosphorylation; Physically Handicapped; Post-Translational Protein Processing; Principal Investigator; Process; Protein Analysis; Protein Binding; Proteins; Proteomics; Public Health; Receptor Cell; Recombinants; Recurrence; Resistance; Role; Serum; Signal Pathway; Signal Transduction; Site; Staging; Structure; Surface; Surface Antigens; Techniques; Toll-Like Receptor 2; Trypanosoma cruzi; United States; Urine; Vaccines; Vesicle; Virulence Factors; base; cytokine; effective therapy; glycosylation; immune resistance; immunoregulation; insight; macrophage; member; migration; parasite invasion; pathogen; programs; receptor; research study; response; tandem mass spectrometry; trans-sialidase

DESCRIPTION: Trypanosoma cruzi causes Chagas' disease, which affects over 11 million people in Latin America. Of those infected, about 5 million will develop severe cardiac and/or digestive disorders. Annually, up to 50,000 people may die and many others will become physically disabled. Recently, Chagas' disease became a public health concern in the United States, owing to the rising number of chronically infected migrants. Currently, there is only one partially effective drug commercially available; there is no human vaccine. Our long-term goal is to understand the molecular events involved in the interaction of T. cruzi with host cells, aiming at the establishment of rational bases for the development of more effective therapies Our hypothesis is that alphaGal-containing vesicles shed by the infective trypomastigote stage of T. cruzi (TcalphaGalVes) contain the major parasite virulence factors, responsible for both the recurrent cell invasion and escaping from host immunity. We also propose that TcalphaGalVes might be accountable for the marked inflammatory process observed in the chronic phase of the disease. Our hypothesis is based on the observations that TcalphaGalVes greatly enhance the host cell invasion by engaging Toll-like receptor 2 (TLR2). Our specific aims are: Specific Aim # 1: To determine the molecular composition of TcalphaGalVes. We will perform a detailed analysis of proteins and post-translational modifications (PTMs), such as glycosylation, glycosylphosphatidylinositol (GPI) anchoring, and phosphorylation. Specific Aim # 2: To define how TcalphaGalVes interact with host cell receptors and enhance cell invasion. We intend to identify and characterize the host alphaGal-binding protein, study its interaction with TLR2, and learn how this leads to the increase of parasite entry into the cell. Electron and confocal microscopy analyses of the early interactions between TcalphaGalVes and host cells will be carried out. We believe that the successful achievement of these specific aims will greatly advance our knowledge of the fine molecular mechanisms used by T. cruzi to invade host cells and escape from the host anti-parasitic immunity. Our ultimate goal is to establish a rational basis for the development of more effective therapies against this deadly pathogen.

描述：克氏锥虫引起恰加斯病，影响拉丁美洲超过 1100 万人。在感染者中，约 500 万人将出现严重的心脏和/或消化系统疾病。每年，可能有多达 50,000 人死亡，还有许多人身体残疾。最近，由于慢性感染的移民人数不断增加，恰加斯病成为美国的一个公共卫生问题。目前，市面上只有一种部分有效的药物；没有人类疫苗。我们的长期目标是了解克氏锥虫与宿主细胞相互作用所涉及的分子事件，旨在为开发更有效的疗法奠定合理基础。我们的假设是，感染性锥鞭毛体脱落的含有 alphaGal 的囊泡克氏锥虫 (TcalphaGalVes) 阶段含有主要的寄生虫毒力因子，负责反复细胞入侵和逃避宿主免疫。我们还提出 TcalphaGalVes 可能与疾病慢性期观察到的明显炎症过程有关。我们的假设基于这样的观察：TcalphaGalVes 通过与 Toll 样受体 2 (TLR2) 结合，极大地增强了宿主细胞的侵袭。我们的具体目标是： 具体目标#1：确定 TcalphaGalVes 的分子组成。我们将对蛋白质和翻译后修饰 (PTM) 进行详细分析，例如糖基化、糖基磷脂酰肌醇 (GPI) 锚定和磷酸化。具体目标#2：确定 TcalphaGalVes 如何与宿主细胞受体相互作用并增强细胞侵袭。我们打算鉴定和表征宿主 αG​​al 结合蛋白，研究其与 TLR2 的相互作用，并了解这如何导致寄生虫进入细胞的增加。将对 TcalphaGalVes 和宿主细胞之间的早期相互作用进行电子和共聚焦显微镜分析。我们相信，这些具体目标的成功实现将极大地增进我们对克氏锥虫入侵宿主细胞并逃避宿主抗寄生虫免疫的精细分子机制的了解。我们的最终目标是为开发针对这种致命病原体的更有效疗法奠定合理的基础。