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Mycobacterium avium subsp. paratuberculosis (MAP) during bovine monocyte-derived macrophage infection: untreated vs. bafilomcyin A1 treatment

ABSTRACT: Transcriptional profiling of M. avium subsp. paratuberculosis inside bovine monocyte-derived macrophages (MDMs; in vitro infection) during phagosome acidification at 30 min post-infection. M. avium subsp. paratuberculosis profiles from bafilomcyin A1 (acidification inhibitor) treated and untreated monocyte-derived macrophages were compared. Two-condition experiment; 3 control (bafilomcyin treated) and 3 test, independently conducted. 1 control, 1 test per microarray.

ORGANISM(S): Mycobacterium avium subsp. paratuberculosis

SUBMITTER: Elise Lamont

PROVIDER: E-GEOD-84708 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Mycobacterium avium subsp. paratuberculosis (MAP) during bovine monocyte-derived macrophage infection: untreated vs. bafilomcyin A1 treatment

Project description:Transcriptional profiling of M. avium subsp. paratuberculosis inside bovine monocyte-derived macrophages (MDMs; in vitro infection) during phagosome acidification at 30 min post-infection. M. avium subsp. paratuberculosis profiles from bafilomcyin A1 (acidification inhibitor) treated and untreated monocyte-derived macrophages were compared.

2016-07-29 | GSE84708 | GEO

Expression analysis of Mycobacterium avium subsp. paratuberculosis K-10 within macrophage cell line

Project description:Once entering the cell, M. avium subsp.paratuberculosis is known to survive harsh microenvironments, especially those inside activated macrophages. To improve our understanding of M. avium subsp.paratuberculosis pathogenesis, we examined the phagosome maturation associated with transcriptional responses of M. avium subsp.paratuberculosis during macrophage infection. Monitoring cellular markers, only live M. avium subsp.paratuberculosis bacilli were able to prevent phagosome maturation and reduce its acidification. On the transcriptional level, over 300 of M. avium subsp.paratuberculosis genes were significantly, differentially regulated in both naM-CM-/ve and IFN-M-NM-3-activated macrophages. These genes include the sigma factor H (sigH) that was shown to be important during persistent infection in M. tuberculosis. Bacterial total RNA was purified from Mycobacterium avium subsp. paratuberculosis that was infected into J774A.1 cells grown with or without interferon-gamma activation at 2 or 24 h post-infection. RNA from bacteria incubated with PRMI-1640 medium for 2 h, which was the infection conditon, is used as control condition. Each condition has two biological replicates. Each hybridization represents expression levels of all 4,350 annotated genes with 20 60-mer oligonucleotides and with three technical replicates.

2013-01-22 | E-GEOD-43645 | biostudies-arrayexpress

Global host transcriptional profiles following Mycobacterium avium subsp. paratuberculosis infection in mice

2014-10-31 | GSE62836 | GEO

Global host transcriptional profiles following Mycobacterium avium subsp. paratuberculosis infection in mice

Project description:We focused on how Mycobacterium avium subsp. paratuberculosis influences the subsequent host response to investigate the host immunopathology accompanying the host anti-mycobacterial immune response during Mycobacterium avium subsp. paratuberculosis infection in spleen of mice. We analyzed altered transcription in the spleen of mice at 3, 6, and 12 weeks following Mycobacterium avium subsp. paratuberculosis infection.

2014-10-31 | E-GEOD-62836 | biostudies-arrayexpress

Expression analysis of Mycobacterium avium subsp. paratuberculosis K-10 within macrophage cell line

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The Intracellular Life of Mycobacterium avium subsp. paratuberculosis During Macrophage Stress: A Key Role for the Alternative Sigma Factor, SigH

Project description:Mycobacterium avium subspecies paratuberculosis (a.k.a. M. paratuberculosis) causes JohneM-bM-^@M-^Ys disease, an enteric infection in cattle and other ruminants, greatly afflicting the dairy industry worldwide. Once inside the cell, M. paratuberculosis is known to survive harsh microenvironments, especially those inside activated macrophages. To improve our understanding of M. paratuberculosis pathogenesis, we examined the phagosome maturation associated with transcriptional responses of M. paratuberculosis during macrophage infection. Monitoring cellular markers, only live M. paratuberculosis bacilli were able to prevent phagosome maturation and reduce its acidification. On the transcriptional level, over 300 of M. paratuberculosis genes were significantly, differentially regulated in both naM-CM-/ve and IFN-M-NM-3-activated macrophages. These genes include the sigma factor H (sigH) that was shown to be important during persistent infection in M. tuberculosis. Interestingly, a sigH-knockout mutant showed increased sensitivity to a sustained level of thiol-specific oxidative stress. Large-scale RNA sequence analysis revealed that a large number of genes belong to the sigH regulon, especially following diamide stress. Genes involved in oxidative stress and virulence were among the induced genes in the sigH regulon with a putative consensus sequence for SigH binding was recognized in a subset of these genes (N=30), suggesting direct regulation with SigH. Finally, mice infections showed a significant attenuation of the M-bM-^HM-^FsigH mutant compared to its parental strain suggesting a role for sigH in M. paratuberculosis virulence. Such analysis could identify potential targets for further testing as vaccine candidates against JohneM-bM-^@M-^Ys disease. Examination of role played by alternative sigma factor, SigH during M. paratuberculosis infection.

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The Intracellular Life of Mycobacterium avium subsp. paratuberculosis During Macrophage Stress: A Key Role for the Alternative Sigma Factor, SigH

Project description:Mycobacterium avium subspecies paratuberculosis (a.k.a. M. paratuberculosis) causes Johne’s disease, an enteric infection in cattle and other ruminants, greatly afflicting the dairy industry worldwide. Once inside the cell, M. paratuberculosis is known to survive harsh microenvironments, especially those inside activated macrophages. To improve our understanding of M. paratuberculosis pathogenesis, we examined the phagosome maturation associated with transcriptional responses of M. paratuberculosis during macrophage infection. Monitoring cellular markers, only live M. paratuberculosis bacilli were able to prevent phagosome maturation and reduce its acidification. On the transcriptional level, over 300 of M. paratuberculosis genes were significantly, differentially regulated in both naïve and IFN-γ-activated macrophages. These genes include the sigma factor H (sigH) that was shown to be important during persistent infection in M. tuberculosis. Interestingly, a sigH-knockout mutant showed increased sensitivity to a sustained level of thiol-specific oxidative stress. Large-scale RNA sequence analysis revealed that a large number of genes belong to the sigH regulon, especially following diamide stress. Genes involved in oxidative stress and virulence were among the induced genes in the sigH regulon with a putative consensus sequence for SigH binding was recognized in a subset of these genes (N=30), suggesting direct regulation with SigH. Finally, mice infections showed a significant attenuation of the ∆sigH mutant compared to its parental strain suggesting a role for sigH in M. paratuberculosis virulence. Such analysis could identify potential targets for further testing as vaccine candidates against Johne’s disease.

2012-12-29 | GSE43182 | GEO

Data-dependent acquisition based proteomic analysis of Mycobacterium avium subsp. paratuberculosis ATCC 19698

Project description:Detection of species-specific proteotypic peptides for accurate and easy characterization of infectious non-tuberculous mycobacteria such as Mycobacterium avium subsp. paratuberculosis is essential. Therefore, we conducted an in-depth global proteomic experiment using M. avium subsp. paratuberculosis ATCC 19698 (Map) strain followed by proteome database search and spectral library generation. The lysate was subjected to in-solution proteomic sample preparation and fractionated using an offline C18 StageTip. Each fraction was acquired in technical triplicates using a 180 min data-dependent acquisition (DDA) method in Orbitrap Fusion Tribrid (Thermo Scientific) mass spectrometer. The resulting raw DDA data were searched against the M. avium subsp. paratuberculosis proteome database using Proteome Discoverer and FragPipe. The resulting peptide spectrum matches were converted into a spectral library using BiblioSpec.

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Transcriptional profiling of whole-blood in Mycobacterium avium subsp. paratuberculosis-infected cattle

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