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Transcriptional responses of murine macrophages to the adenylate cyclase toxin of Bordetella pertussis

ABSTRACT: Three different recombinant forms of CyaA were used to investigate transcriptional responses of murine bone marrow-derived macrophages (BMDMs) using Affymetrix Mouse Genome Genechips. These forms were enzymically active, invasive CyaA, nonenzymically active, invasive CyaA (CyaA*) and non-enzymically active, non-invasive CyaA (proCyaA*). BMMs, treated with 20 ng/ml of CyaA for 24 h, showed over 1000 significant changes in gene transcription compared with control cells. CyaA caused an increase in transcription of many inflammatory genes and genes associated with various signalling cascades such as those involved in cyclic AMP-dependent protein kinase A signalling. Most strikingly, CyaA caused down-regulation of numerous genes involved in cell proliferation. CyaA* at 20 ng/ml significantly up-regulated the transcription of only twelve genes after 24 h whereas proCyaA* at this concentration significantly increased the transcription of only two genes.

ORGANISM(S): Mus musculus

PROVIDER: GSE51075 | GEO | 2013/09/22

SECONDARY ACCESSION(S): PRJNA221120

REPOSITORIES: GEO

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Transcriptional responses of murine macrophages to the adenylate cyclase toxin of Bordetella pertussis

Project description:Three different recombinant forms of CyaA were used to investigate transcriptional responses of murine bone marrow-derived macrophages (BMDMs) using Affymetrix Mouse Genome Genechips. These forms were enzymically active, invasive CyaA, nonenzymically active, invasive CyaA (CyaA*) and non-enzymically active, non-invasive CyaA (proCyaA*). BMMs, treated with 20 ng/ml of CyaA for 24 h, showed over 1000 significant changes in gene transcription compared with control cells. CyaA caused an increase in transcription of many inflammatory genes and genes associated with various signalling cascades such as those involved in cyclic AMP-dependent protein kinase A signalling. Most strikingly, CyaA caused down-regulation of numerous genes involved in cell proliferation. CyaA* at 20 ng/ml significantly up-regulated the transcription of only twelve genes after 24 h whereas proCyaA* at this concentration significantly increased the transcription of only two genes. The effect of administration of three different recombinant forms of pertussis toxin to mouse bone marrow derived macrophages are compared with a vehicle only contol 24 hours after treatment in technical triplicate measurements

2013-09-22 | E-GEOD-51075 | biostudies-arrayexpress

RNA sequencing for enteroids treated with different concentrations of IL-17 (0 ng/ml. 1 ng/ml, 10 ng/ml and 100 ng/ml)

Project description:RNA sequencing for enteroids treated with different concentrations of IL-17 (0 ng/ml. 1 ng/ml, 10 ng/ml and 100 ng/ml)

| PRJNA756286 | ENA

Phosphoproteomics of cAMP signaling of adenylate cyclase toxin in mouse dendritic cells

Project description:The whooping cough agent, Bordetella pertussis, subverts dendritic cell (DCs) functions through powerful immunomodulatory activities of its toxins. Here we focused on the signaling action of the adenylate cyclase toxin (CyaA) that targets myeloid cells expressing the αMβ2 integrin CD11b/CD18 (known as complement receptor 3 (CR3) or Mac-1). CyaA delivers an extremely catalytically potent adenylyl cyclase enzyme domain into the cytosol of phagocytes and disrupts their innate and adaptive immune functions through unregulated production of the key signaling molecule cAMP. Here we describe the global phosphoproteomic analysis of cAMP signaling in murine bone marrow-derived DCs exposed to CyaA. Gathered data reveal toxin-triggered alternations of phosphorylation status of proteins regulating actin cytoskeleton, chromatin remodeling, mTOR activity and IL-10 gene expression. The reported findings highlight the complexity of subversive physiological manipulation that is exerted on myeloid phagocytes by the cAMP-generating adenylate cyclase toxin of Bordetellae.

2017-11-29 | PXD004733 | Pride

TGF-ß treatment of human melanoma cell lines

Project description:To analyse whether the activation of TGF-ß signalling (recombinant hTGF-ß, 5 ng/mL; PeproTech) is able to invoke a full phenotype- switch, we performed RNA sequencing of melanoma cultures from two independent melanoma cell lines at two different time point after TGF-β1 (5 ng/ml, preprotech).

2020-04-17 | GSE148767 | GEO

IgG-independent co-aggregation of FceRI and FcgRIIB results in LYN- and SHIP1-dependent tyrosine phosphorylation of FcgRIIB

Project description:We used tyrosine phosphorylation profiling by anti-pTyr-antibody mediated enrichment and subsequent analysis by mass spectrometry in order to determine the changes in early signalling after mast cell activation. Mast cells were stimulated with varying concentrations of antigen for one minute and the differences between optimal (20 ng/ml) and supra-optimal (2000 ng/ml) antigen mast cell activation were analysed by nanoLC-MS/MS.

2018-08-17 | PXD009318 | Pride

Retargeting from CR3 to the LFA-1 receptor uncovers the adenylyl cyclase enzymetranslocating segment of Bordetella adenylate cyclase toxin

Project description:The Bordetella adenylate cyclase toxinhemolysin (CyaA) and the α-hemolysin (HlyA) of Escherichia coli both belong to the family of cytolytic pore-forming Repeats in ToXin (RTX) cytotoxins. HlyA preferentially binds the αLβ2 integrin LFA-1 (CD11a/CD18) of leukocytes and can promiscuously bind and permeabilize also a variety of other cells. CyaA bears an Nterminal adenylyl cyclase enzyme (AC) domain linked to a pore-forming RTX cytolysin (Hly) moiety and binds the complement receptor 3 (CR3, αMβ2, CD11b/CD18 or Mac-1) of myeloid phagocytes, penetrates their plasma membrane and delivers into cytosol the AC enzyme. We constructed a set of CyaA/HlyA chimeras and show that the CyaC-acylated segment and the CR3-binding RTX domain of CyaA can be functionally replaced by the much shorter HlyCacylated and LFA-1-binding RTX moiety of HlyA. Instead of binding CR3, a CyaA1710/HlyA411-1024 chimera bound the LFA-1 receptor and effectively delivered the AC enzyme into Jurkat lymphoblastoma T cells. At high chimera concentrations (25 nM), the interaction with LFA-1 was not required for CyaA1-710/HlyA411-1024 binding to CHO cells. However, interaction with the LFA-1 receptor strongly enhanced the specific capacity of the bound CyaA1-710/HlyA411-1024 chimera to penetrate cells and deliver the AC enzyme into their cytosol. Hence, interaction of the acylated RTX moiety of HlyA with LFA-1 promoted a productive membrane interaction of the chimera. These results allow to delimit the residues 400 to 710 of CyaA as the 'AC translocon' sufficient for translocation of the AC enzyme polypeptide across the plasma membrane of target cells

2020-05-11 | PXD018789 | Pride

Global gene expression profiling of FOP- or resFOP-iMSCs treated by several ligands

Project description:Analyzed differentially expressed genes among FOP- or resFOP-iMSCs treated by several ligands: Activin-A, 100 ng/mL; BMP-7, 100 ng/mL; TGF-B3, 10 ng/mL

2015-12-09 | GSE62783 | GEO

Real-time quantitative PCR analysis of human dendritic cells

Project description:Human CD14 positive monocytes were purified from healthy volunteers' blood and were differentiated to immature dendritic cells in vitro by culturing for five days in the presence of interleukin-4 (IL-4 100 ng/ml) and GM-CSF (75 ng/ml). Immature dendritic cells were activated three different ways for 24 hours: 1. Double-stranded DNA poly(dA:dT) (2.5 μg/ml) complexed with LyoVec transfection reagent. 2. LPS (500 ng/ml) 3. Inflammatory cocktail containing 10 ng/ml TNF, 5 ng/ml IL-1β, 20 ng/ml IL-6, 75 ng/ml GM-CSF and 1 μg/ml PGE2. We used SA Biosciences Antigen Presenting and Toll-like Receptor Pathway PCR Arrays to quantitate gene expression of immunologically relevant genes from the immature and the differently activated cells.

2011-05-08 | GSE29131 | GEO

Bulk RNAseq of AML and endothelial cells in untreated and cytokine-treated conditions

Project description:We report the use of bulk RNAseq of AML and endothelial cell lines in four conditions (1. Untreated; 2. IFNg - 10 ng/ml; 3. TNFa - 10 ng/ml; 4. IFNg and TNFa - 10 ng/ml) in order to identify genes that are differentially expressed between the two cell types, both at baseline and in an inflammatory microenvironment.

2021-10-21 | GSE159991 | GEO

RNA sequencing for enteroids treated with different concentrations of IL-17 (0 ng/ml. 1 ng/ml, 10 ng/ml and 100 ng/ml)

Project description:To evaluate the intestinal epithelial responses induced by IL-17, bulk RNA-sequencing (RNA-seq) was performed on the human small intestinal organoids (enteroids, n = 3) treated with different concentrations of IL-17 (0 ng/ml. 1 ng/ml, 10 ng/ml and 100 ng/ml) .

2021-08-20 | GSE182453 | GEO

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