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The In Vitro Interaction of Aspergillus fumigatus and Human immature Dendritic Cells

ABSTRACT: Dendritic cells (DC) play an important role in host immunity by acting as a bridge between the innate and adaptive immune systems. They are antigen presenting cells that obtain microbial antigens by direct phagocytosis of the microbe or by cross presentation of antigens taken up from the surrounding environment. Monocyte derived DC were co-cultured with resting conidia of Aspergillus fumigatus at an MOI of 5 for 12 hours, cells were sampled every three hours. RNA was extracted from both organisms at each time point and hybridised to micro-arrays, whole genome Aspergillus fumigatus array (JCVI) and a custom immune array for DC. The genes up-regulated by DC in the presence of A. fumigatus indicated that the cells were producing a pro-inflammatory response. There was an increase in IL8 expression over time confirming its association with germ tube emergence. Over the course of the experiment there was increased expression of 210 genes by A. fumigatus, GO analysis indicated significant up-regulation of the following biological processes: fermentation, drug transport, pathogenesis, transport, tyrosine catabolism and response to oxidative stress. There were two clusters of temporally regulated genes showing up regulation before 6hr and after 6 hrs. This may be related to the increased mortality exhibited in DC at 6h. The initial analysis of A. fumigatus gene expression in response to DC shows similarity to its response to neutrophils with an up-regulation in catabolism and response to oxidative stress.

ORGANISM(S): Aspergillus fumigatus Aspergillus fumigatus Af293

PROVIDER: GSE22053 | GEO | 2010/07/01

SECONDARY ACCESSION(S): PRJNA128973

REPOSITORIES: GEO

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The In Vitro Interaction of Aspergillus fumigatus and Human immature Dendritic Cells

Project description:Dendritic cells (DC) play an important role in host immunity by acting as a bridge between the innate and adaptive immune systems. They are antigen presenting cells that obtain microbial antigens by direct phagocytosis of the microbe or by cross presentation of antigens taken up from the surrounding environment. Monocyte derived DC were co-cultured with resting conidia of Aspergillus fumigatus at an MOI of 5 for 12 hours, cells were sampled every three hours. RNA was extracted from both organisms at each time point and hybridised to micro-arrays, whole genome Aspergillus fumigatus array (JCVI) and a custom immune array for DC. The genes up-regulated by DC in the presence of A. fumigatus indicated that the cells were producing a pro-inflammatory response. There was an increase in IL8 expression over time confirming its association with germ tube emergence. Over the course of the experiment there was increased expression of 210 genes by A. fumigatus, GO analysis indicated significant up-regulation of the following biological processes: fermentation, drug transport, pathogenesis, transport, tyrosine catabolism and response to oxidative stress. There were two clusters of temporally regulated genes showing up regulation before 6hr and after 6 hrs. This may be related to the increased mortality exhibited in DC at 6h. The initial analysis of A. fumigatus gene expression in response to DC shows similarity to its response to neutrophils with an up-regulation in catabolism and response to oxidative stress. A. fumigatus AF293 cells were grown in the presence and absence of human dendritic cells for 0h - 12h. Hybridizations were performed with biological replicates and flip-dye pairs.

2010-07-01 | E-GEOD-22053 | biostudies-arrayexpress

Response of A549 cells treated with Aspergillus fumigatus

Project description:This SuperSeries is composed of the following subset Series: GSE24983: Response of A549 cells treated with Aspergillus fumigatus [WT-CF_vs_WT-GC] GSE24984: Response of A549 cells treated with Aspergillus fumigatus [WT-GC_vs_PrtT-GC] GSE24985: Response of A549 cells treated with Aspergillus fumigatus [WT-CF_vs_PrtT-CF] Refer to individual Series

2010-10-29 | E-GEOD-24986 | biostudies-arrayexpress

NFAT-dependent IL-2 and IL-23 produced by DC are key regulators of pathogenic (TH17-mediated lung inflammation)

Project description:Calcineurin/NFAT/IL-2 signaling pathway is activated in dendritic cells (DC) upon encounter of Î² glucan, the main component of the fungal cell wall, raising the question about the role of NFAT-regulated genes in DC biology in vivo. To directly assess the function of IL-2 secreted by DC, we analyzed mice lacking of IL-2 in the DC lineage, CD4-expressing cells and with complete deletion of IL-2 in the germ line in a mouse model of pulmonary fungal infection. Here we found that specifically the loss of IL-2 in DC resulted in increased mice mortality upon the fungus Aspergillus fumigatus challenge and expansion of Th17 cells in the lung. We demonstrated that only CD103+DC were able to release IL-2 in acute phase of pulmonary Aspergillosis through the Ca2+-Calcineurin-NFAT signaling. We also found that NFAT mediates IL-23 transcription in lung DC, where IL-2 results essential in restraining the priming of a pathogenic infiltrating IL-17+Sca1+CD90+CD4+ cell with stem cell like properties. Thus, IL-2 and IL-23 secreted by DC in the lung have an antagonistic relationship on the Th17 differentiation program with IL-2 inducing T cell differentiation and IL-23 inducing a stem cell like molecular signature to Th17 cells upon Aspergillus challenge. DC-Il2-/- then confer the Th17 stemness, releasing IL-23 in response to the fungus contributing to the development of a Th17 cell effector population, particularly pathogenic in infection. D1 cells with no treatment, or treatment with different fungal types or antigens at 1, 4 and 6 hours, in triplicate, with the 3 untreated samples at 1hr also including techincal repeats

2015-12-21 | E-GEOD-58590 | biostudies-arrayexpress

Transcriptional profiling of Aspergillus fumigatus in response to RBL-2H3 mast cells

Project description:In patients with chronic pulmonary disease colonization with the mold Aspergillus fumigatus is associated with declining pulmonary function and obstructive airway disease. One potential effector of this inflammatory response is the pulmonary mast cell. In vitro studies have demonstrated that A. fumigatus contact induces IgE-independent mast cell degranulation. Conversely, the Aspergillus secondary metabolite gliotoxin has been shown to suppress mast cell activation. These contradictory results emphasize the need for a better understanding of the interactions between A. fumigatus and mast cells. Thus, the objective of this work was to identify A. fumigatus genes that are differentially regulated upon exposure to mast cells. Transcriptional profiling experiments indicated that, in addition to genes encoding for iron acquisition systems, allergens and putative virulence factors, genes from the gliotoxin biosynthesis cluster were significantly down-regulated upon exposure to mast cells. Globally, the results from this study provide insight into the A. fumigatus response to mast cells and suggest that one mechanism by which the host may circumvent the effects of gliotoxin is via the suppression of fungal gliotoxin synthesis by mast cells.

2012-03-02 | GSE27610 | GEO

RNA-seq analysis of alleles of the atrR transcription factor-encoding gene in Aspergillus fumigatus

Project description:Determine the transcriptomic response of Aspergillus fumigatus strains to different alleles of atrR

2019-02-11 | GSE123445 | GEO

The fungus Aspergillus niger consumes sugars in a sequential manner that is not mediated by the carbon catabolite repressor CreA

Project description:Aspergillus niger is a filamentous ascomycete fungus that is commonly found in most biotopes around the globe. In nature, A. niger degrades the plant biomass polysaccharides to monomeric sugars, transports them into the cells, and uses a variety of catabolic pathways to convert them into biochemical building blocks and energy. We show that when grown in liquid cultures, A. niger takes up plant-biomass derived monomeric sugars (and maltose) in a highly sequential manner, rather than simultaneously. Interestingly, this sequential uptake was not mediated by the fungal general carbon catabolite repressor protein CreA, which has been shown to mediate the use of preferential carbon sources over non-preferential carbon sources. Furthermore, transcriptome analysis strongly indicated that the preferential use of the monomeric sugars is arranged at the level of transport, but it is not reflected in transcriptional regulation of sugar catabolism. Therefore, the results indicate that the regulation of sugar transport and catabolism are separate physiological processes in A. niger.

2018-04-23 | GSE98434 | GEO

Comparative genomic hybridization of Aspergillus fumigatus Af71, Aspergillus fumigatus Af294, Aspergillus clavatus, Neosartorya fenneliae and Neosartorya fischeri with Aspergillus fumigatus Af293

Project description:Genomic DNA from five strains, Aspergillus fumigatus Af71, Aspergillus fumigatus Af294, Aspergillus clavatus, Neosartorya fenneliae, and Neosartorya fischeri, were co-hybridized with that of Aspergillus fumigatus Af293 and compared.

2005-05-15 | E-MEXP-333 | biostudies-arrayexpress

Differential gene expression in human myeloid dendritic cells interacting with different knockout strains of Aspergillus fumigatus in vitro

Project description:Dendritic cells (DC) are the most important antigen presenting cells and play a pivotal role in host immunity to infectious agents by acting as a bridge between the innate and adaptive immune systems. Myeloid DCs (mDC) were isolated from peripheral blood and co-cultured with conidia (con) and germlings (ks) of Aspergillus fumigatus knockout (mitA, mnt1, rodA) and wildtype (wt) strains for 6 hours at an MOI of 1. RNA was extracted and hybridised to microarrays. Microarrays contain probes for approximately 110 selected human immune-relevant genes, including cytokine and chemokine genes, their receptors and downstream innate immunity genes. We used microarrays to detail the gene expression of human myeloid dendritic cells after 6h of co-cultivation with Aspergillus fumigatus wildtype and knockout mutants

2012-04-16 | E-GEOD-34884 | biostudies-arrayexpress

Gene expression profile of HIF-1α silenced or non-silenced human DCs stimulated with A. fumigatus under normoxic or hypoxic conditions

Project description:DCs play a central role for the immune response against the mold Aspergillus fumigatus. Hypoxic microenvironments occur during infection with A. fumigatus. Hypoxia and signaling via hypoxia inducible factor 1α may modulate the response of DCs; however, the role in fungal infections is unclear. We used microarrays to determine the influence of hypoxia and HIF-1α signaling on the immune response of human DCs towards A. fumigatus.

2015-11-01 | GSE60729 | GEO

Long-term adaptation to hypoxic conditions - Febit transcriptome analysis

Project description:Aspergillus fumigatus is an opportunistic, airborne pathogen causing invasive aspergillosis in immunocompromised patients. During the infection process A. fumigatus is challenged by hypoxic microenvironments occurring in inflammatory, necrotic tissue. To gain further insights into the adaptation mechanism, A. fumigatus was cultivated in an oxygen-controlled chemostat under hypoxic and normoxic conditions. Transcriptome analysis revealed significant increases in transcripts associated with cell wall polysaccharide metabolism, amino acid and metal ion transport, nitrogen metabolism and glycolysis. A concomitant reduction in transcript levels was observed with cellular trafficking and G-protein coupled signaling. To learn more about the functional roles of hypoxia-induced transcripts we deleted A. fumigatus genes putatively involved in reactive nitrogen species detoxification (fhpA), NAD+ regeneration (frdA, osmA) nitrogen metabolism (niaD, niiA) and respiration (rcfB). We show that the NO-detoxifying flavohemoprotein fhpA is strongly induced by hypoxia independent of the nitrogen source, but is dispensable for hypoxic survival. By deleting the nitrate reductase gene niaD, the nitrite reductase gene niiA and the two fumarate reductases genes frdA and osmA, we found that alternative electron acceptors such as nitrate and fumarate do not have a significant impact on growth of A. fumigatus during hypoxia, but that functional mitochondrial respiratory chain complexes are essential under these conditions. Inhibition studies indicated that primarily complex III and IV play a crucial role in the hypoxic growth of A. fumigatus.

2014-07-01 | E-MTAB-2699 | biostudies-arrayexpress

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