Clade specific virulence patterns of M. tuberculosis complex strains in human macrophages: Molecular mechanisms underlying phagosomal escape and induction of cell death.

结核病的转归取决于结核分枝杆菌的毒力和耐药性。结核菌复合体（MTC）菌株和宿主巨噬细胞的杀伤能力。在目前的研究中，我们将研究病原体的变异性和宿主细胞信号转导的变异性的MTC菌株窝藏室，吞噬体的分子组成的影响。该研究包括（i）比较分析来自人原代巨噬细胞的含有MTC进化枝I和进化枝II的吞噬体的蛋白质和脂质组成，（ii）鉴定控制M.结核菌在巨噬细胞中的生长，以及（iii）从人和鼠巨噬细胞分离的吞噬体的差异分析，以定义在两个物种中存在并参与的“核心蛋白质组组分”和在两个物种中的一个中存在或专门募集到吞噬体的“个体蛋白质组组分”。一种新的脂质为基础的程序来标记细菌表面和快速免疫磁性技术将被用来分离完整的含菌室从小鼠和人类的原代细胞，以进一步表征它们的电子和原子力显微镜，流式细胞术，蛋白质印迹和质谱。新开发的方法有助于细胞内病原体王国的比较研究，并将提供给优先计划的其他成员。

The outcome of tuberculosis is determined by both the virulence of the infecting M. tuberculosis complex (MTC) strain and the killing capacity of the host macrophage. In the current study we will examine the impact of pathogen variability and host cell signaling variability on the molecular composition of the MTC strain harboring compartment, the phagosome. The study comprises (i) the comparative analysis of the protein and lipid composition of MTC clade I and clade II containing phagosomes from human primary macrophages, (ii) the identification of critical host cell factors that govern control of M. tuberculosis growth in macrophages and (iii) the differential analysis of phagosomes isolated from human and murine macrophages to define “core proteome components” which are present and engaged in both species and “individual proteome components” which are present or exclusively recruited to the phagosome in one of the two species. A new lipid-based procedure to label bacterial surfaces and a rapid immunomagnetic technique will be used to isolate intact bacteria-containing compartments from mouse and human primary cells in order to further characterize them by electron and atomic force microscopy, flow cytometry, Western blot, and mass spectrometry. The newly developed method facilitates comparative studies across the kingdom of intracellular pathogens and will be made available to other members of the priority program.