Project description:Whole cell lysates of THP-1 macrophages infected with of M. tuberculosis CDC1551, a ppe38-ppe71 knock out mutant and a complemented strain was performed at 4h and 18h post infection to find the differentially regulated proteins.
Project description:I infected 8 flasks of resting murine bone marrow derived macrophages with M. tuberculosis CDC1551. Individual samples of intracellular Mtb were collected at 2hr, and days 2,4,6,8,10,12, and 14 post-infection using the GTC (guanidine thiocyanate) method as previously published. A 2hr "no macrophage" control was used as a common denominator in the two-color hybridizations. I have two independent biological replicates of this experiment. Groups of assays that are related as part of a time series. Time: culture time post-infection
Project description:I infected 8 flasks of resting murine bone marrow derived macrophages with M. tuberculosis CDC1551. Individual samples of intracellular Mtb were collected at 2hr, and days 2,4,6,8,10,12, and 14 post-infection using the GTC (guanidine thiocyanate) method as previously published. A 2hr "no macrophage" control was used as a common denominator in the two-color hybridizations. I have two independent biological replicates of this experiment. Groups of assays that are related as part of a time series. Time: culture time post-infection time_series_design
Project description:CDC1551 induced more vigorous immune response in murine bone marrow derived macrophage (BMM). In contrast, in HN878-infected cells, host transcriptional response was delayed but lasted longer. HN878 induced more genes involved in host lipid metabolism than CDC1551 did. Murine bone marrow derived macrophages were infected with M. tuberculosis CDC1551 or HN878 up to 24 h. Total RNA was processed for microarray and global gene expression was read.
Project description:The aim of this study was to identify the signaling pathways differentially engaged upon infection with either live or heat killed Mycobacterium tuberculosis in murine bone marrow derived macrophages. Based on preliminary data that type I IFN signaling dominates the transcriptional differences, we investigated what roles the type I IFN receptor IFNAR and the signaling molecule STING play during infection with either live or heat killed Mycobacterium tuberculosis in bone marrow derived macrophages. In WT and STING KO macrophages, IFNbeta treatment was added either by itself or in addition to both of the infection conditions to test whether the lack of IFNbeta induction in these cells accounted for any differences in transcriptional respones.added IFNbeta treatment either by itself or in addition to both of the infection conditions.
Project description:Transcriptional profiling of Mycobacterium tuberculosis mRNA enriched from host-pathogen samples from in vitro bone marrow derived macrophages (Mus musculus).
Project description:This study uses microarray analyses to examine transcriptional responses of Mycobacterium tuberculosis complex clinical isolates to phagosomal cues encountered inside activated (IFN-gamma+LPS) murine bone marrow-derived macrophages 24hr post-infection. Transcript levels of intracellular mycobacteria were compared to extracellular bacteria of the same strain (An aliquot of the inoculum used to infect macrophages was incubated in the absence of macrophages for 24hr in an identical flask). Set of arrays that are part of repeated experiments Keywords: Biological Replicate Biological Replicate Computed
Project description:This study uses microarray analyses to examine transcriptional responses of Mycobacterium tuberculosis complex clinical isolates to phagosomal cues encountered inside resting murine bone marrow-derived macrophages 24hr post-infection. Transcript levels of intracellular mycobacteria were compared to extracellular bacteria of the same strain (An aliquot of the inoculum used to infect macrophages was incubated in the absence of macrophages for 24hr in an identical flask). Set of arrays that are part of repeated experiments Keywords: Biological Replicate Biological Replicate Computed
Project description:Macrophages play an essential role in the early immune response to Mycobacterium tuberculosis and are the cell type preferentially infected in vivo. Primary macrophages and macrophage-like cell lines are commonly used as infection models, although the physiological relevance of cell lines, particularly for host-pathogen interaction studies, is debatable. Here we use high-throughput RNA-sequencing to analyse transcriptome dynamics of two macrophage models in response to M. tuberculosis infection. Specifically, we study the early response of bone marrow-derived mouse macrophages and cell line J774 to infection with live and -irradiated (killed) M. tuberculosis. We show that infection with live bacilli specifically alters the expression of host genes such as RSAD2, IFIT1/2/3 and RIG-I, whose potential roles in resistance to M. tuberculosis infection have not yet been investigated. In addition, the response of primary macrophages is faster and more intense than that of J774 cells in terms of number of differentially expressed genes and magnitude of induction/repression. Our results point to potentially novel processes leading to immune containment early during M. tuberculosis infection, and support the idea that important differences exist between primary macrophages and cell lines, which should be taken into account when choosing a macrophage model to study host-pathogen interactions.
Project description:We investigated the molecular mechanisms by which colchicine regulates host defense in macrophages against M. tuberculosis infection. To elucidate this, we performed global gene expression analysis of M. tuberculosis infected-bone marrow-derived macrophages after colchicine treatment.
