Leishmania major promastigote entry of the of the autophagic compartment and amastigote escape from the parasitophorous vacuole

利什曼原虫主要前鞭毛体进入自噬室和无鞭毛体从寄生液泡逃逸

• 批准号: 197967827
• 负责人: Professor Dr. Paul Walther
• 金额: --
• 依托单位: Zentrale Einrichtung Elektronenmikroskopie
• 依托单位国家: 德国
• 项目类别: Priority Programmes
• 财政年份: 2011
• 资助国家: 德国
• 起止时间: 2010-12-31 至 2019-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/197967827?language=en
• 关键词: Leishmania; major; promastigote; entry; autophagic

During the first three years we will (i) visualize the leishmania major containing parasitophorous vacuoles in human macrophages using state of the art high pressure freezing and electron tomography focusing on promastigote interactions with autophagy and amastigote escape from the parasitophorous vacuoles (ii) we will try to down regulate autophagy and lysosomal maturation in macrophage cell lines and primary macrophages (iii) we will establish a new assay for correlative electron microscopy. During the second 3-year funding period (i) we aim to dissect the molecular basis leading to promastigote entry of the autophagic pathway and amastigote escape. (ii) We will apply the correlative electron microcopy technique on new target proteins coming from our mass spectrometry screen. In addition we will test L. donovani known to block phagolysosomal maturation and L. amazonensis known to induce large parasitophorous vacuoles. (iii) Moreover we will focus on the parasitophorous vacuoles in human neutrophils that can only be infected with promastigotes and dendritic cells that can only be infected with amastigotes.

在前三年中，我们将（i）使用最先进的高压冷冻和电子断层扫描技术，观察人类巨噬细胞中含有寄生性空泡的利什曼原虫，重点关注前鞭毛体与自噬的相互作用以及无鞭毛体从寄生性空泡中逃逸（ii）我们将尝试下调巨噬细胞系和原代巨噬细胞中的自噬和溶酶体成熟 (iii)我们将建立一种新的关联电子显微镜检测方法。在第二个 3 年资助期 (i)，我们的目标是剖析导致前鞭毛体进入自噬途径和无鞭毛体逃逸的分子基础。 (ii) 我们将把相关电子显微镜技术应用于来自我们的质谱筛选的新目标蛋白。此外，我们将测试已知可阻止吞噬溶酶体成熟的杜氏乳杆菌和已知可诱导大寄生泡的亚马逊乳杆菌。 (iii) 此外，我们将重点关注只能被前鞭毛体感染的人类中性粒细胞中的寄生液泡和只能被无鞭毛体感染的树突状细胞。