Project description:C3heB/FeJ mice were infected with M. tuberculosis to form necrotic granulomatous lesions. FFPE samples of infected lungs with granulomas were microdissected into three distinct regions, Caseum, foamy macrophage, and Cell. Proteins were extracted from microdissected samples, followed by LC-MS/MS.
Project description:The study aims at understanding the global nature of the immune responses within the the lung tissue 28 days post infection with Mycobacterium tuberculosis. Comparing the whole transcriptome of infected lungs to that of an uninfected lungs revealed a plethora of immune mechanisms driven by various cytokines. IFN-γ, IL-6, IL-2, TNFα were the major cytokines observed. We observed significant differential expression of gene involved in the JAK-STAT and the MAPK signalling pathways. We observed an interplay of the immune regulatory genes and various non-immune genes controlling the metabolism, apoptosis, cell growth, post translational modifications etc.
Project description:To gain insight into the host cell types, cellular and molecular pathways possibly involved in the differential permissiveness to pulmonary replication of M. tuberculosis, we carried out transcript profiling studies on M. tuberculosis-infected lungs from congenic and parental strains. We were particularly interested in two groups of transcripts. The first group consists of transcripts which expression in the lung is regulated in response to M. tuberculosis infection (global response to infection), and that is obtained by comparing transcripts profiles of infected vs. uninfected lungs. The second group of transcripts is associated with increased resistance to M. tuberculosis infection of B6 and D2.B6-Chr7 mice. That list consists in the overlap between the lists commonly expressed in response to infection between resistant B6 and D2.B6-Chr7 but that show a significant difference in modulation when compared to infected susceptible D2.<br><br> In these experiments, B6, D2 as well as the D2.B6-Chr19, and D2.B6-Chr7 congenic lines were infected with M. tuberculosis and lungs were harvested at day 30 and day 70, and RNA was prepared. Three independent RNA samples from each group were converted to labeled cDNAs and hybridized to Affymetrix oligonucleotides arrays (Mouse Genome 430 2.0 array). Hybridization results were analyzed with the Genesifter analysis program to characterize changes in gene expression.
Project description:Define genes differentially expressed by Nur77-GFP HI and LO CD4 T cells FACS sorted from the lungs of Mycobacterium tuberculosis-infected mice
Project description:Treatment of chronic inflammatory diseases with tumor necrosis factor alpha antagonists has been associated with increased risk of tuberculosis (TB). We examined the usefulness of the rabbit model of active pulmonary TB for studying the impact of the human immune modulatory reagent etanercept on the host immune response. Control of Mycobacterium tuberculosis (Mtb) infection, disease pathology and the global transcriptional response in Mtb-infected lungs of rabbits were studied. Etanercept treatment exacerbated disease pathology and reduced bacillary control in the lungs, compared to infected untreated animals. The microarray experiments involves comparison of changes in gene expression between Mtb-HN878 infected and Etanercept treated and untreated rabbit lungs at 4 and 8 weeks post-treatment, starting at 4 weeks post-infection.
Project description:Define genes differentially expressed by Nur77-GFP HI and LO CD4 T cells FACS sorted from the lungs of Mycobacterium tuberculosis-infected mice with bulk RNA seq
Project description:Using whole genome microarrays, we compared changes in gene expression patterns in the lungs of TB-resistant A/Sn and TB-susceptible I/St mice at day 14 following infection with Mycobacterium tuberculosis H37Rv. Analyses of differentially expressed genes for representation of gene ontology terms and activation of regulatory pathways revealed interstrain differences in antigen presentation, NK, T and B cell activation pathways. In general, resistant A/Sn mice exhibited a more complex pattern and stronger activation of host defense pathways compared to the TB-susceptible I/St mouse strain. In addition, in I/St mice elevated activation of genes involved in neutrophil response was observed and confirmed by quantitative RT-PCR and histopathology. Furthermore, a specific post infection upregulation of cysteine protease inhibitors was found in susceptible I/St mice. To monitor the global transcriptome patterns in the two mouse strains, gene expression was assessed using Agilent 4 x44K whole genome microarrays. Genes that changed their expression level at least two-fold compared to non-infected controls with a p-value of 0.1 were considered as differentially expressed genes (DEG).
Project description:We sought to develop and characterize a novel paucibacillary model in mice, which develop necrotic lung granulomas following infection with Mycobacterium tuberculosis. Paucibacillary infection was established, recapitulating the sterilizing activities of human LTBI regimens. TNF neutralization led to increased lung bacillary load, disrupted granuloma architecture with expanded necrotic foci and reduced tissue hypoxia, and accelerated animal mortality. TNF-neutralized mouse lungs and sera showed significant upregulation of IFN?, IL-1?, IL-6, IL-10, CCL2, CCL3, and matrix metalloproteinase genes Six weeks after aerosol-immunization with recombinant M. bovis BCG overexpressing the 30-kilodalton antigen, C3HeB/FeJ mice were aerosol-infected with M. tuberculosis H37Rv. Six weeks later, mice were treated with one of three standard regimens for latent TB infection (LTBI) or TNF-neutralizing antibody. Mouse lungs were analyzed by histology, positron emission tomography/computed tomography, whole-genome microarrays, and RT-PCR. Lungs and sera were studied by multiplex enzyme-linked immunosorbent assays