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THP-1 cells infected with the clinical isolates of extensively drug-resistant and drug sensitive tuberculosis

ABSTRACT: Extensively drug resistant tuberculosis (XDR-TB) showed many different characteristics including the extreme drug resistance versus the drug sensitive clinical isolates (DS-TB), to know better about the reasons we used the tuberculosis host cells named as THP-1 (one kind of the macrophage cells) to be infected by the XDR-TB and DS-TB.DS strain A36 and the XDR strain B42 and was typical and selected by our lab. Then the total RNA of infected or uninfected THP-1 cells was extract and purified for the analysis by the chip (22K Human Genome chip representing the 21522 ORF of human with the oligonucleotide probe of 70 mer from CapitalBio Corp., Beijing, China). The results reflected the different expressed genes involved in apoptosis, secreted cytokines and signal pathway and so on. Those results might indicate the how the XDR-TB cause the pathogenesis. In this study, the well grown THP-1 cells were separated and cultured in three ampoules. Cells in one ampoules were infected with XDR-TB strain of B42. Cells in another ampoules were infected with DS-TB strain of A36, with the cells in the third one were not infected and just treated with PBS as the control. Then the dual channel method was used for detecting the hybridization of B42 vs the control or A36 vs control. This work was repeated for three times.

ORGANISM(S): Homo sapiens

SUBMITTER: Yi Liu

PROVIDER: E-GEOD-15539 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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THP-1 cells infected with the clinical isolates of extensively drug-resistant and drug sensitive tuberculosis

Project description:Extensively drug resistant tuberculosis (XDR-TB) showed many different characteristics including the extreme drug resistance versus the drug sensitive clinical isolates (DS-TB), to know better about the reasons we used the tuberculosis host cells named as THP-1 (one kind of the macrophage cells) to be infected by the XDR-TB and DS-TB.DS strain A36 and the XDR strain B42 and was typical and selected by our lab. Then the total RNA of infected or uninfected THP-1 cells was extract and purified for the analysis by the chip (22K Human Genome chip representing the 21522 ORF of human with the oligonucleotide probe of 70 mer from CapitalBio Corp., Beijing, China). The results reflected the different expressed genes involved in apoptosis, secreted cytokines and signal pathway and so on. Those results might indicate the how the XDR-TB cause the pathogenesis.

2009-04-04 | GSE15539 | GEO

Expresson from human THP-1 stimulated with IFN-g and/or infected with TB

Project description:Investigation of interacting effects of TB and IFN-g on gene expression regulation within human macrophage-like THP-1 cells To investigate the effect TB infection has on THP-1 macrophageâ??s response to IFN-g post-infection, in multiple experiments, THP-1 cells were exposed to each of the following four conditions: not treated, IFN-Î³ treated, M. tuberculosis infected and M. tuberculosis infected then IFN-Î³ treated. Total cellular RNA was then extracted. Recovered RNAs were pre-qualified for reproducibility by comparing qRT-PCR results of IFIT-2 and IP10 between replicate experiments. Four most reproducible replicate experiments were then selected for genome-wide mRNA abundance assays using Affymetrix microarray-based technology.

2010-03-09 | E-GEOD-17477 | biostudies-arrayexpress

Changes in the membrane proteome of exosomes released from human macrophages after infection with Mycobacterium tuberculosis.

Project description:One of the greatest obstacles to eliminating TB is the lack of predictive and diagnostic biomarkers. Exosomes are a promising alternative source of biomarkers for TB since they can carry mycobacterial antigens. We hypothesize that exosomes from Mtb-infected macrophages exhibit a characteristic selection of human protein that can be evaluated as TB biomarkers. We looked for the localization—exosomal membrane or lumen—of the differentially abundant proteins. Exosomes were obtained from THP-1-derived macrophages (Mtb infected and uninfected controls). The exosome population was validated by nanoparticle tracking analysis and western blot. The protein composition of exosomes was evaluated by tandem mass spectrometry. Differences in protein composition and abundances between exosomes from infected and control cells were evaluated by t-test the proteomics findings were confirmed by western blot. Using a biotinylation strategy we verified the protein localization. Forty-seven proteins were significantly more abundant in exosomes from Mtb-infected cells, 66% were predicted to be membrane associated. The biotinylation pattern confirmed the membrane-association of some of these proteins.

2016-12-20 | PXD004062 | Pride

Changes in the Membrane-Associated Proteins of Exosomes Released from Human Macrophages after Mycobacterium tuberculosis Infection - Biotinylated

Project description:Tuberculosis (TB) is the deadliest infectious disease worldwide. One obstacle hindering the elimination of TB is our lack of understanding of host-pathogen interactions. Exosomes, naturally loaded with microbial molecules, are circulating markers of TB. Changes in the host protein composition of exosomes from Mycobacterium tuberculosis (Mtb)-infected cells have not been described, can contribute to our understanding of the disease process, and serve as a direct source of biomarkers or as capture targets to enrich for exosomes containing microbial molecules. Here, the protein composition of exosomes from Mtb-infected and uninfected THP-1-derived macrophages was evaluated by tandem-mass-spectrometry and differences in protein abundances were assessed. Our results show that infection with Mtb leads to significant changes in the protein composition of exosomes. Specifically, 41 proteins were significantly more abundant in exosomes from Mtb-infected cells; 63% of these were predicted to be membrane associated. Thus, we used a novel biotinylation strategy to verify protein localization, and confirmed the localization of some of these proteins in the exosomal membrane. Our findings reveal another important scenario where Mtb could be influencing changes in host cells that unveil new features of the host-pathogen interaction and may also be exploited as a source of biomarkers for TB.

2018-08-06 | PXD010659 | Pride

Transcriptomics of vitamin D treatment effects in Mycobacterium tuberculosis-infected THP-1 cells

Project description:Previous reports have shown low vitamin D serum levels and polymorphisms in the vitamin D receptor (VDR) to be associated with increased risk for TB. Given that 1?,25-dihydroxyvitamin D3 has a role in lipid metabolism control, we tested whether the link between 1?,25-dihydroxyvitamin D3 and tuberculosis involves macrophage lipid metabolism. Since formation of lipid droplets (LD) is a hallmark of lipid dysregulation in M. tuberculosis-infected macrophages, we measured LD content as a readout of altered lipid metabolism in infected THP-1 cells. Induction of LD, which peaked by 24 hours post-infection was prevented by addition of 1?,25-dihydroxyvitamin D3 at the time of infection. To investigate the mechanism of 1?,25-dihydroxyvitamin D3 modulation of LD formation, we analyzed the transcriptome of M. tuberculosis-infected THP-1 cells with and without 1?,25-dihydroxyvitamin D3 treatment. THP-1 cells were cultured in 24-well flat bottom plates, pre-treated with 20ng/ml PMA for 24 h, and incubated with M. tuberculosis H37Rv. After 4 hours of infection, cells were washed to remove extracellular bacteria and treated with 100 nM of 1?,25-dihydroxyvitamin D3. THP-1 cells were collected and analyzed at 24h. 3 biological replicates in each of the 4 groups were prepared: Infected, Infected plus 1?,25-dihydroxyvitamin D3, Non-infected, Non-infected plus 1?,25-dihydroxyvitamin D3.

2014-07-01 | E-GEOD-57028 | biostudies-arrayexpress

Transcription profiling of human cell line THP-1 cells exposed to Daboiatoxin and cytokines involved in melioidosis caused by B. pseudomalli

Project description:Gene expression profiles of human cell (THP-1) lines exposed to a novel Daboiatoxin (DbTx) isolated from Daboia russelli russelli, and specific cytokines and inflammatory pathways involved in acute infection caused by Burkholderia pseudomallei. Experiment Overall Design: 1. Group I:- Human monocytic macrophage (THP-1) cell lines grown in the culture medium without any bacterial infection served as untreated control group (Three Biological Replicates). Experiment Overall Design: 2. Group II:- THP-1 cells were infected with Burkholderia pseudomallei (A600 nm = OD 0.6, ~5 x 107 cfu/ml) for 24h served as a disease control group (Three Biological Replicates). Experiment Overall Design: 3. Group III:- THP-1 cells were infected with B. pseudomallei and treated with Daboiatoxin (0.5 mM) isolated from Daboia russelli russelli venom served as a treatment group (Three Biological Replicates). Experiment Overall Design: 4. Group IV:- THP-1 cells were infected with B. pseudomallei (A600 nm = OD 0.6, ~5 x 107 cfu/ml) treated with standard antimicrobial drug ceftazidime (10mg/ml) served as a drug control (Three Biological Replicates). Experiment Overall Design: 5. Group V:- THP-1 cells were exposed to Daboiatoxin (0.5 mM) without bacterial infection (Three Biological Replicates).

2009-04-04 | E-GEOD-7577 | biostudies-arrayexpress

MtrA is involved in progression of Mycobacterium tuberculosis cell cycle

Project description:Synchronised cultures of Mycobacterium tuberculosis (M. tb H37Ra DnaAcos115) were used to determine the cell cycle dependent gene expression. Mycobacterium tuberculosis H37Ra DnaAcos115 strain was grown in Middlebrook 7H9 broth at 30C for 30h which arrests replication and later cultures were allowed to grow at 37oC for 30h. Samples were collected at 0, 2, 6, 12,18, 24 and 30 hours. Samples at 30oC for 30h were considered as '0' time point. A total of 18 samples, consisting of triplicates for 6 time-points were analyzed using microarray.

2015-12-31 | E-GEOD-53640 | biostudies-arrayexpress

Monocyte-like THP-1 Cells: Trichosporon asahii Infected vs Control

Project description:Transcriptional profiling of Monocyte-like THP-1 Cells with Trichosporon asahii Infected THP-1 Cells comparing untreated THP-1 cells Two-condition experiment,Trichosporon asahii Infected THP-1 Cells vs THP-1 Cells. Biological replicates: 3 infected THP-1 Cells, independently grown and harvested.One replicate per array.

2012-11-03 | E-GEOD-41983 | biostudies-arrayexpress

Metabolomic analysis of Mycobacterium tuberculosis reveals metabolic profiles for identification of drug-resistant tuberculosis

Project description:Metabolomics of 150 Mtb isolates (54 pre-XDR, 63 XDR-TB and 33 pan-susceptible; pan-S)

2023-02-23 | MSV000091354 | MassIVE

Expression analysis of Mycobacterium tuberculosis H37Rv biofilm after treatment with DTT ( Reduced) for 29 hrs

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

2016-02-08 | E-GEOD-75847 | biostudies-arrayexpress

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