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

最近，对吞噬体摄取乳胶后的蛋白质组分析促进了我们对这些隔室的生物学的理解。对含有吞噬体的病原体的类似分析迄今为止受到污染细胞组分的影响，这是蛋白质组学研究的严重障碍。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。应制备表达内吞途径的特定囊泡群体的荧光标记物的转基因小鼠。来自不同靶组织的原代宿主细胞感染了不同标记的病原体，然后将允许分离细胞内栖息地。标记的标记物限定了吞噬体成熟中可区分的步骤，因此即使在可能的异步感染的情况下，也应该允许一定的动力学解析。作为应用，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L.墨西哥。

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.