MiRNA profiles of lung tissues from Mtb H37Rv infected C57B/6 mice and uninfected controls
ABSTRACT: The C57B/6 mice were infected with Mtb H37Rv (CFU=200) for 28 days, and the miRNA expression profile from lung tissues were analyzed with deep sequencing. Overall design: miRNA profiles of lung tissues from Mtb H37Rv infected C57B/6 mice and uninfected controls were generated by deep sequencing, in triplicate.
Tuberculosis (TB) caused by Mycobacterium tuberculosis (Mtb) kills millions every year, and there is urgent need to develop novel anti-TB agents due to the fast-growing of drug-resistant TB. Although autophagy regulates the intracellular survival of Mtb, the role of calcium (Ca2+) signaling in modulating autophagy during Mtb infection remains largely unknown. Here, we show that microRNA miR-27a is abundantly expressed in active TB patients, Mtb-infected mice and macrophages. The target of miR-27 ...[more]
Project description:The peritoneal macrophages were infected with Mtb H37Rv for 4 hours, and the miRNA expression profile were analyzed with deep sequencing. Overall design: miRNA profiles of Mtb H37Rv infected peritoneal macrophages and uninfected controls were generated by deep sequencing, in triplicate.
Project description:Mtb H37Rv was found to be highly susceptible to ATD-3169 (Redox generating compound). To further understand the mechanism of ATD-3169 action, we performed microarray analysis of Mtb H37Rv exposed to 3µM, 30 µM and 150µM of ATD-3169 for 4h. Overall design: Mtb H37Rv was grown till early log phase and treated with ATD-3169 concentrations (3uM, 30uM, 150uM) for 4 hours at 37 degrees at 180rpm. Three independent experiments were done for each of the strain.
Project description:Genomic comparisons among different lineages Mtb isolates. Overall design: Laboratoy strain Mtb H37Rv was compered to nine Mtb clinical isolates of the Beijing lineage. One, two or three replicates were performed for each comparison. Laboratory strain Mtb H37Rv was also compared to two laboratory, genetically-modified derivates from Mtb HN878 clinical isolate in which one copy of the dosR gene was inactivated. Two replicates were performed for each comparison.
Project description:Transcriptional profiling of RAW264.7 cells infected with M. tuberculosis H37Rv at an MOI of 10 performed 4 and 24 hours post-infection. RAW264.7 cells were infected with Mtb for 4 hours at an MOI of 10. Cells were washed and treated with gentamycin for 2 hours in order to remove adhered bacteria. Incubation was continued further for 4 and 24 hours followed by total RNA isolation. Total RNA was labelled with Agilent’s quick-Amp labelling kit (p/n: 5190-0444) to generate fluorescent complementary RNA by using T7 promoter based linear amplification. The control sample was labelled with Cy3 while the infected samples were labelled with Cy5 and hybridized to an Agilent oligo microarray kit.
Project description:Since WhiB3 expression was shown to be maximally upregulated at pH 4.5 and MtbΔwhiB3 was shown to be defective in survival in Ph 4.5, we did microarray at pH 4.5 so as to study comprehensive role of WhiB3 in regulating gene expression at acidic Ph Overall design: Mtb H37Rv and MtbΔwhiB3 were grown till early log phase and exposed to 7H9-Tylaxapol at pH 4.5 for 2 hours at 37 degree at 150rpm. Two independent experiments were done for each of the strain.
Project description:Amox-Clav combination downregulates the expression of WhiB4. Further, MtbΔwhiB4 survives better upon treatment with this combination. Therefore, to study the role of WhiB4 in regulating the response of Mtb to this combination, we carried out global transcriptome profiling of Wt Mtb and MtbΔwhiB4 strains after treatment. Overall design: Mtb H37Rv and MtbΔwhiB4 were grown till early log phase and treated with 10X Augmentin at 100 µg/ml of Amoxicillin and 8 µg/ml of Clavulanate for 6 h at 37 degree at 150rpm. Wildtype Mtb H37Rv were treated with 1X Augmentin (10 µg/ml of Amoxicillin and 8 µg/ml of Clavulanate), or 5X Augmentin (50 µg/ml of Amoxicillin and 8 µg/ml of Clavulanate) for 6 or 12 h at 37 degree at 150rpm. Experiments were done in duplicate, triplicate, tetraplicate or pentaplicate.
Project description:Transcriptional profiling of H37Rv and ∆mtrB strains grown to log phase in MB7H9 Overall design: Comparison of ∆mtrB transcriptional profile with H37Rv as control. Two technical replicates per strain, one experiment.
Project description:Expression profile of Mycobacterium tuberculosis H37Rv biofilm as induced by DTT (Reduced) 6mM DTT reduced at 6 mM concentration was added to log phase culture of Mtb H37Rv. After 29 hours RNA was isolated and hybridization was done on microarrays
Project description:Tuberculosis kills nearly 2 million people through out the world, every year. The outcome of M. tuberculosis infection is determined by the host and bacterial factors. A strong host immune response controls the growth of the bacilli effectively. However, in a host with suboptimal immune response, the bacilli grows and mounts disease. Activation of immune response following M. tuberculosis infection affects the expression of many host genes that are involved in the production of immune system molecules such as cytokines, chemokines, surface receptors and transcriptional regulators that manifest in the change of subsequent cellular events, including chemotaxis and proliferation of effector cells. The infecting bacilli counteracts the bactericidal activities of the host immune cells, primarily by modifying their gene expression. However, the specific nature of the host-pathogen interactions and the outcome of Mtb infection are not fully understood. Tumor necrosis factor-alpha (TNF-a), produced by the immune cells, is an important cytokine in protecting the host against Mtb infection. However, excessive TNF-a production leads to severe inflammation and host cell destruction. In fact, pharmacologic inhibition of TNF-a production has been considered as a therapeutic modality in inflammatory diseases. Interestingly, inhibitors of host phosphodiesterase-4 (PDE4) have been shown to reduce TNF-a production and dampen inflammation without complete immune suppression of the host. In this study, we have explored the use of one of the PDE4 inhibitors, CC-3052, as an adjunct immune modulatory drug, in combination with isoniazid (INH) treatment in rabbit pulmonary tuberculosis. We hypothesize that reducing TNF-a levels during Mtb infection would reduce the environmental pressure on the bacteria, rendering them more amenable to killing by INH. Mtb infected rabbits were treated with CC-3052 or INH or both and the lung tissue were harvested after 4 and 8 weeks of treatment. Lung bacterial load, histologic changes and host and bacterial gene expression were determined for each timepoint and compared between various treatment groups. The results of our study provides data to support the idea that combining anti-TB drugs with an adjunctive immune modulator may enhance the efficacy of current TB therapy regimens and shorten the duration of treatment if applied appropriately to humans. The microarray experiments involves 2 comparison groups. 1) Changes in rabbit gene expression between Mtb infected and uninfected animals; 2). Changes in rabbit gene expression between CC-3052 treated and untreated animals during Mtb infection at 4,8 and 12 weeks post infection. New Zealand White rabbits were infected with Mtb HN878 at 3.2log10 (on day 0). At 4 weeks post infection, one group of infected rabbits were treated with a phosphodiesterase-4 (PDE4) inhibitor, CC-3052, and the treatment was continued up to 12 weeks post infection. The compound was used at 25mg/kg body weight, dissolved in distilled water and administered through gavage five days a weeks. Lung tissue from Uninfected, Mtb-infected and Untreated or CC-3052 treated rabbits were isolated at Day0, 4,8 and 12 weeks post infection and used for total RNA extraction. Only the 8 and 12 week timepoints correspond to the associated publication.