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

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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.