Project description:Analysis of Metfromin induced changes in the lung cells of Mycobacterium tuberculosis infected mouse at gene expression level. The hypothesis tested in the present study was whether metformin has any effect on the host immune response in Mycobacterium tuberculosis infected mice? Results provide important information on the effect of metformin on the inflammatory response and immune activation associated with mycobacterial infection. In conclusion, Metfromin normalizes the chronic inflammation associated with Mycobacterium tuberculosis infection.
Project description:Paraffin-embedded lung and spleen tissues analyzed by Eksigent nanoLC-Ultra 2D System and QExactive mass spectrometer. Both lung and spleen tissues were extracted from animals at 4 different conditions (Not infected Ad libitum, Not infected Caloric restricted, Mycobacterium Tuberculosis (MTB) infected Ad libitum, Mycobacterium Tuberculosis (MTB) infected Caloric restricted). Globally, 24 and 23 runs are uploaded for lung and spleen tissues, respectively.
Project description:Neutrophils are accumulated in mouse lungs following Mycobacterium tuberculosis infections. Increased influx of neutrophils is associated with increased susceptibility to disease, but the underlying mechanisms of neutrophil mediated Tuberculosis disease pathogenesis is poorly understood. To understand the transcriptomic differences between resistant (C57 BL/6) and susceptible (C3HeB/FeJ) mouse lung neutrophils, we sorted them at Day 29 post infection and analyzed them by bulk mRNA sequencing.
Project description:Mycobacterium tuberculosis(Mtb) is known to reside in cells of innate immune system- macrophages and dendritic cells. A variety of non -conventional cell typeslike adipocytes, mesenchyal stem cells and osteoclasts can also be infected with Mtb. However, cellular transcriptional adapations enabling survival of Mtb in these cells remain known. We used microarrays to understand global changes in transcriptional profiling during macrophage to osteoclast transition in presence of Mycobacterium tuberculosis.
Project description:Tuberculosis (TB), caused by Mycobacterium tuberculosis (Mtb), is defined by granulomas—immune aggregates that either contain or support bacterial replication. Macrophages, fundamental components of these lesions, are crucial to TB pathogenesis, yet their phenotypic and functional diversity remains incompletely understood. Here, we used single-cell RNA sequencing and immunofluorescence to profile macrophages in lung tissue and granulomas from a nonhuman primate model of early TB. We identified transcriptionally distinct subsets, including embryonic-origin tissue-resident alveolar macrophages and monocyte-derived alveolar and interstitial macrophages, each with distinct spatial localization in granulomas. Tissue-resident alveolar macrophages and a subset undergoing epithelial-to-mesenchymal transition accounted for the highest frequency of Mtb-infected cells. Infected cells exhibited differential expression of immune- and migration-associated genes compared to uninfected counterparts, suggesting Mtb either induces or exploits these pathways as a survival strategy. These findings highlight macrophage heterogeneity as a major driver of differential susceptibility to Mtb and provide insights relevant to future immunomodulatory strategies.
Project description:Pyrazinamide (PZA) is one of the first line antibiotics used for the treatment of tuberculosis (TB). we have used human monocyte and a mouse model of pulmonary TB to investigate whether treatment with PZA, in addition to its known anti-mycobacterial properties, modulate the host immune response during Mycobacterium tuberculosis (Mtb) infection. Mice were infected with Mtb and treatment with PZA was started at 28 days post-infection. At 42 days and 63 days post-infection, group of animals were euthanized and lung tissue was collected to isolate total RNA and used in microarray experiments. Mtb-infected, untreated animals served as controls.
Project description:Infection by Mycobacterium tuberculosis (Mtb) continues to cause significant mortality due to pathogen persistence in lung cells derived from blood monocytes. Since persistent blood monocytosis is a hallmark of tuberculosis and monocyte-derived lung subsets differ in their ability to restrict the growth of intracellular Mtb in mice, the ontogeny of these subsets could inform development of host-directed therapies. Recent literature suggests that circulating monocytes are heterogeneous and derive from mobilization of distinct bone marrow (BM) or spleen progenitors that direct local differentiation. However, the role of the inflamed lung environment on this process has not been addressed. We compared monopoiesis in the BM of Mtb-infected mice and uninfected controls and found that these monocyte pools had similar differentiation within the infected lung. While Mtb infection induced splenic monopoiesis, we found no impact on lung monocyte differentiation in splenectomized mice. However, when monocytes were transferred into Mtb-infected recipients with a distinct lung environment, we observed altered differentiation. We identified monocyte-derived lung subsets with unique gene expression, associated with specific spatial distributions and cell neighborhoods. These findings suggest that the local lung environment has a larger influence on the phenotypic diversity of monocyte-derived lung cells than does the peripheral environment.