Project description:Mycobacterial transcripts were identified in exosomes released from M.tb infected RAW264.7 macrophages that were not present in uninfected exosomes suggesting export of mycobacterial RNA via exosomes Mycobacterial RNA was used as positive control and RNA from exosomes released from uninfected macrophages was used as negative control
Project description:Investigation to study mRNA transcripts present in exosomes from M.tb infected cells and how they compare to those derived from uninfected cells. Transcripts were also studied in donor macrophages as controls The gene expression study identified unique transcripts as well as differentially expressed transcripts present in exosomes released from infected macrophages
Project description:we studied the role of exosomes isolated from M.tb infected macrophages in modulating the macrophage response to IFN-γ. Nimblegen microarray gene expression studies were used to compare the suppression of IFN-γ inducible genes by exosomes relative to the virulent strain of M.tuberculosis. Overall our study suggest that exosomes, as carriers of M.tb pathogen associated molecular patterns (PAMPs), may provide a mechanistic link by which M.tb may exert its suppression of host immune response beyond the infected cell, and implies a physiological role for exosomes in immune surveillance of TB. Macrophages were treated with exosomes, infected with M.tb H37Rv or left untreated for 18 hours followed by +/- IFN-γ for an additional 18 hours. Cells were harvested and RNA was isolated and converted to double stranded cDNA and subsequently labeled and hybridized onto Mus musculus 4×72 Nimblegen microarray using Nimblegen Hybridization system 4 according to manufacturer’s instructions (Roche)
Project description:we studied the role of exosomes isolated from M.tb infected macrophages in modulating the macrophage response to IFN-γ. Nimblegen microarray gene expression studies were used to compare the suppression of IFN-γ inducible genes by exosomes relative to the virulent strain of M.tuberculosis. Overall our study suggest that exosomes, as carriers of M.tb pathogen associated molecular patterns (PAMPs), may provide a mechanistic link by which M.tb may exert its suppression of host immune response beyond the infected cell, and implies a physiological role for exosomes in immune surveillance of TB.
Project description:We infect bone marrow derived macrophages (BMDMs) with either a Hyper or Hypovirulent Mycobacterium tuberculolsis strain and assess the host response to these bacteria through RNAseq Uninfected BMDMs served as controls, BMDMs infected with Hypervirulent M.tb (designated R55), BMDMs infected with Hypovirulent m.tb (designated R50). 3 groups with 3 biological replicates, each analysed in triplicate.
Project description:We infect bone marrow derived macrophages (BMDMs) with Mycobacterium cultured with and without Tween 80 and asses the host response to these bacteria through RNAseq Uninfected BMDMs served as controls, BMDMs infected with detergent-free M.tb (designated RNT), BMDMs infected with Tween cultured m.tb (designated RT). 3 groups with 3 biological replicates, each analysed in triplicate.
Project description:In this study, RNA sequencing (Transcriptome sequencing) was employed to investigate the global transcriptome changes in the macrophages during the different strains of M.tb infection. THP-1 cells derived macrophages were exposed to the virulent M.tb strain H37Rv (Rv) or the avirulent M.tb strain H37Ra (Ra), and the M.tb BCG vaccine strain was used as a control. The cDNA libraries were prepared from M.tb infected macrophages and then sequenced.
Project description:Extracellular vesicles (EVs) released by Leishmania donovani infected RAW264.7 murine macrophages were evaluated for their proteomic composition and potential function in pathogenesis. EVs were collected after 72 hours infection and analyzed by LC-MS/MS using both in gel and in solution digestion approaches to identify protein composition of both host and parasite origin.