Functional characterization of cell-specific differences of L. major-containing phagolysosomes in macrophages and dendritic cells

在被沙蝇叮咬传播到皮肤后，利什曼原虫主要原生动物寄生虫被巨噬细胞（MF）和树突状细胞（DC）吞噬。感染期promastigote L.的主要生命形式由皮肤寄生MF占据，在此期间，寄生虫在含有寄生虫的吞噬溶酶体内转化为专性细胞内生命形式，即寄生液泡（PV）。相反，DC优先摄取无纺丝。这两种吞噬细胞（MF， DC）被寄生虫内化后的细胞内事件尚未得到很好的表征。然而，寄生虫在这些细胞内的命运及其细胞行为存在显著差异，例如，仅在DC中观察到IL-12释放，而在MF中没有。最近，我们发现细胞内吞噬溶酶体TLR9信号对于感染后dc来源的IL-12产生至关重要。因此，我们现在打算进一步研究各自含l - major的pv中存在的成分和条件。利用透射电镜和无偏差的系统生物学方法绘制吞噬囊泡蛋白质组的变化和差异，绘制两种细胞感染后宿主和病原体转录组的变化，我们旨在识别参与吞噬和PV维持的宿主和病原体衍生因子，以及表征细胞类型内遇到的不同环境适应性的病原体衍生因子。

After transmission into the skin by the bite of a sand fly, Leishmania major protozoan parasites are phagocytosed by macrophages (MF), and dendritic cells (DC). The infectious stage promastigote L. major life form is taken up by skin-resident MF, in which the parasite transforms into the obligate intracellular life form within the parasite-containing phagolysosome, the parasitophorous vacuole (PV). In contrast, DC preferentially ingest amastigotes. The intracellular events after parasite internalization by these two phagocytes (MF, DC) are not well characterized. However, the fate of the parasite within these cells and their cellular behaviour differ significantly, e.g. IL-12 release is only observed in DC, but not MF. Recently, we identified intracellular, phagolysosomal TLR9 signalling as critical for DC-derived IL-12 production after infection. Thus, we now intend to further study the constituents and conditions that exist in the respective L. major-containing PVs in more detail. Using TEM and unbiased systems biology approaches to map changes and differences in the proteome of phagocytic vesicles to chart changes in both host and pathogen transcriptomes after infection in the two cell types, we aim to identify host as well as pathogen-derived factors involved in phagocytosis and PV maintenance as well as pathogen-derived factors characterizing adaptation to the different environments encountered inside the cell types.