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

巨噬细胞和树突状细胞中含有 L. Major 吞噬溶酶体的细胞特异性差异的功能表征

• 批准号: 261181825
• 负责人: Professorin Dr. Ruth Esther von Stebut-Borschitz
• 金额: --
• 依托单位: Klinik für Dermatologie und Venerologie
• 依托单位国家: 德国
• 项目类别: Priority Programmes
• 财政年份: 2014
• 资助国家: 德国
• 起止时间: 2013-12-31 至 2016-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/261181825?language=en
• 关键词: Functional; characterization; cell; specific; differences

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.

主要利什曼原虫寄生虫通过白蛉叮咬传播到皮肤后，被巨噬细胞（MF）和树突状细胞（DC）吞噬。感染期前鞭毛体L.主要生命形式被皮肤驻留MF吸收，其中寄生虫转化为含有寄生虫的吞噬溶酶体内的专性细胞内生命形式，即寄生虫空泡（PV）。相反，DC优先摄取无鞭毛体。这两种吞噬细胞（MF，DC）的寄生虫内化后的细胞内事件没有得到很好的表征。然而，寄生虫在这些细胞内的命运及其细胞行为显著不同，例如仅在DC中观察到IL-12释放，而在MF中未观察到。最近，我们确定了细胞内，吞噬溶酶体TLR 9信号转导的DC衍生的IL-12的生产感染后的关键。因此，我们现在打算进一步研究在各个L中存在的成分和条件。主要包含PV更详细。使用TEM和公正的系统生物学方法来映射吞噬囊泡的蛋白质组的变化和差异，以绘制两种细胞类型感染后宿主和病原体转录组的变化，我们的目标是确定宿主以及病原体衍生的因子参与吞噬和PV维持以及病原体衍生的因子表征适应细胞类型内遇到的不同环境。