Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Monocyte-derived macrophages stimulated with the TLR8 agonist 3M-002 at 10 microM show a prominent activation of TLR-dependent genes as assessed b "Human Toll-like Receptor PCR array pathway" over time

ABSTRACT: We were interested in the effects of TLR8 triggering of human monocyte-derived macrophages. We observed a striking upregulation of those genes. This work was done in the context of examining the effects of TLR8 triggering for the activation of HIV. We generated monocyte-derived macrophages (MDM) by initially isolating monocytes and subsequently culturing them in RPMI supplemented with human AB serum. MDM were stimulated for 18 h with the TLR8 agonist 3M-002, and subsequently harvested for extraction of total RNA. Total RNA was then used to quantify the TLR-dependent genes using a PCR array pathway.

ORGANISM(S): Homo sapiens

SUBMITTER: Roberto Speck

PROVIDER: E-GEOD-26470 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

TLR8 activates HIV from latently infected cells of myeloid-monocytic origin directly via the MAPK pathway and from latently infected CD4+ T cells indirectly via TNF-α.

Schlaepfer Erika E Speck Roberto F RF

Journal of immunology (Baltimore, Md. : 1950) 20110228 7

We previously showed that the TLR7/8 agonist, R-848, activated HIV from cells of myeloid-monocytic origin. In this work, we show that this effect was solely due to triggering TLR8 and that NF-κB was involved in the TLR8-mediated activation of HIV from latently infected cells of myeloid-monocytic origin. Inhibition of Erk1/2 or p38α resulted in attenuation of TLR8-mediated activation of NF-κB. Western blots confirmed that TLR8 triggering activated Erk1/2 and p38α but, surprisingly, not JNK. Altho ...[more]

PMID: 21357269

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Project description:Background: Mycobacterium avium subspecies paratuberculosis (MPTb) is the causative agent of Johne’s disease, an intestinal disease of ruminants with major economic consequences. MPTb bacilli are phagocytosed by host macrophages upon exposure where they persist, resulting in lengthy subclinical phases of infection that can lead to immunopathology and disease dissemination. Consequently, analysis of the macrophage transcriptome in response to MPTb infection can provide valuable insights into the molecular mechanisms that underlie Johne’s disease. Here, we investigate pan-genomic gene expression in bovine monocyte-derived macrophages (MDM) purified from seven age-matched non-related females, in response to in vitro infection with MPTb (multiplicity of infection 2:1) at intervals of 2 hours, 6 hours and 24 hours post-infection. Results: Differentially expressed genes were identified by comparing the transcriptomes of the infected MDM to the control MDM at each time point (adjusted P-value threshold ≤ 0.10). 1,050 differentially expressed unique genes were identified 2 hours post-infection, with 974 and 84 differentially expressed unique genes detected 6 hours and 24 hours post-infection, respectively. Furthermore, in the infected MDM the number of upregulated genes exceeded the number of downregulated genes at each time point, with the fold-change in expression for the upregulated genes markedly higher than that for the downregulated genes. Conclusions: Inspection and systems analysis of the differentially expressed genes revealed changes in the expression profiles of genes involved in the inflammatory response, cell signalling pathways and apoptosis. The transcriptional changes associated with cellular signalling and the inflammatory response may reflect different immuno-modulatory mechanisms that underlie host-pathogen interactions during infection. Affymetrix GeneChip® Bovine Genome Arrays were used to examine gene expression of bovine monocyte-derived macrophages (MDM) after in vitro challenge with Mycobacterium avium subspecies paratuberculosis across a time series of 2 hr, 6 hr and 24 hr post-challenge. A 0 hr control treatment was also generated and seven different age-matched female Holstein-Friesian cattle were used for each time-point/treatment combination for a total of 49 microarrays, one of which was excluded from analysis since it did not pass quality control.

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Dataset Information

Monocyte-derived macrophages stimulated with the TLR8 agonist 3M-002 at 10 microM show a prominent activation of TLR-dependent genes as assessed b "Human Toll-like Receptor PCR array pathway" over time

Publications

TLR8 activates HIV from latently infected cells of myeloid-monocytic origin directly via the MAPK pathway and from latently infected CD4+ T cells indirectly via TNF-α.

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