A coloured approach to study the dynamics and composition of organelles harboring intracellular pathogens such as Leishmania sp..

一种彩色方法，用于研究含有利什曼原虫等细胞内病原体的细胞器的动态和组成。

• 批准号: 5434929
• 负责人: Dr. Anton Aebischer
• 金额: --
• 依托单位: Fachgebiet 16: Erreger von Pilz- und Parasiteninfektionen und Mykobakteriosen
• 依托单位国家: 德国
• 项目类别: Priority Programmes
• 财政年份: 2004
• 资助国家: 德国
• 起止时间: 2003-12-31 至 2006-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/5434929?language=en
• 关键词: coloured; approach; study; dynamics; composition

The proteome analysis of phagosomes after uptake of latex boosted recently our understanding of the biology of these compartments. Similar analyses for pathogen containing phagosomes suffered so far from contaminating cellular components, a serious obstacle for proteomic studies. We aim to develop a flow fluorimetric approach to purify intracellular habitats from primary host cells. Transgenic mice that express fluorescent markers of particular vesicle populations of the endocytic pathway shall be made. Primary host cells from diverse target tissues infected with differently labeled pathogens would then allow to isolate intracellular habitats. the labeled markers define distinguishable steps in phagosome maturation and should therefore allow a certain kinetic resolution, even in the likely case of asynchronous infection. As an application the parasite habitats in dendritic cells (DC) and in macrophages (Mf) from these mice will be compared adopting this approach since we recently noticed a functional difference between these cells due to infection with L. mexicana.

吸收乳胶后吞噬体的蛋白质组学分析最近促进了我们对这些隔室生物学的理解。到目前为止，对含有吞噬体的病原体的类似分析还没有污染细胞成分，这是蛋白质组学研究的一个严重障碍。我们的目标是开发一种流动荧光法来纯化原代宿主细胞的细胞内栖息地。应制作表达内吞途径特定囊泡群荧光标记的转基因小鼠。来自不同目标组织的原代宿主细胞感染了不同标记的病原体，然后可以分离细胞内栖息地。标记的标记物定义了吞噬体成熟的可区分步骤，因此即使在可能的非同步感染情况下，也应该允许一定的动力学分辨率。作为一种应用，我们将采用这种方法比较来自这些小鼠的树突状细胞（DC）和巨噬细胞（Mf）中的寄生虫栖息地，因为我们最近注意到由于感染墨西哥乳杆菌而导致这些细胞之间的功能差异。