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

ABSTRACT: 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.

ORGANISM(S): Homo sapiens

PROVIDER: GSE34884 | GEO | 2012/04/17

SECONDARY ACCESSION(S): PRJNA150205

REPOSITORIES: GEO

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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

The temporal dynamics of differential gene expression in human alveolar epithelial and endothelial cells interacting with the human pathogenic mould Aspergillus fumigatus in vitro

Project description:Aspergillus fumigatus is the most relevant pathogenic mould in immunocompromised patients. Spores of A. fumigatus are inhaled and reach the lung alveoli, where they interact with the human immune system. In this set of experiments, we co-cultured with defined morphologies of Aspergillus fumigatus (Af293) for 0h, 3h and 6h with either cells representing human alveolar epithelium (A549) or human alveolar endothelium (HPAEC). We included either monocyte-derived or myeloid dendritic cells from volunteer donor blood to the epithelial layer to examine their effect on gene expression. RNA was extracted from both cell lines at each time point 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.

2012-04-17 | GSE28806 | GEO

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.

2010-07-01 | GSE22053 | GEO

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

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

The temporal dynamics of differential gene expression in human alveolar epithelial and endothelial cells interacting with the human pathogenic mould Aspergillus fumigatus in vitro

2012-04-16 | E-GEOD-28806 | biostudies-arrayexpress

Small RNA sequencing of Aspergillus fumigatus

Project description:Aspergillus fumigatus is an important human pathogen and a leading fungal killer. This study aimed to determine the small RNA repertoire of A. fumigatus in conidia and mycelium grown for 24 or 48 hours in liquid culture.

2023-02-14 | GSE223619 | GEO

Next Generation Sequencing of HUVECs stimulated by conidia of Aspergillus fumigatus

Project description:Purpose: The goal of this study is to investigate the responses of HUVECs after the stimulation of conidia of A. fumigatus Methods: HUVECs were stimulated with conidia of Aspergillus fumigatus for 2 and 6 hours. Three biological repeats of stimulated cells or un-stimulated controls were send for RNA sequencing. Results: Using an optimized data analysis workflow, we mapped about 40 million sequence reads per sample to the human genome (build hg38) and identified round 80,000 transcripts in the HUVECs upon stimulation. Conclusions: Our resutls showed the detailed analysis of HUVECs transcriptomes upton conidia of Aspergillus fumigatus stimulation.

2022-03-29 | GSE191240 | GEO

Gene expression level of Aspergillus fumigatus CEA17_ΔakuBKU80 at three different stages

Project description:We report the gene expression level of Aspergillus fumigatus CEA17_ΔakuBKU80 strain in dormant conidia, in swollen conidia (after 4h of culture in Glucose 3%, Yeast Exrtract 1% liquid medium) and in germinated conidia (after 8h of culture in Glucose 3%, Yeast Extract 1% liquid medium after construction of an RNAseq library.

2016-09-26 | GSE81956 | GEO

Healthy peripheral neutrophils total RNA-Seq: Serum Amyloid P component is an essential element of resistance against Aspergillus fumigatus

Project description:Serum amyloid P component (SAP; APCS gene, also known as Pentraxin 2, PTX2), is a component of the humoral arm of innate immunity involved in resistance to bacterial infection and regulation of tissue remodeling. The present study was designed to investigate the role of SAP/PTX2 in antifungal resistance. SAP-deficient Apcs-/- mice showed a dramatic increase in susceptibility to A. fumigatus infection. The defective anti-fungal resistance of gene targeted mice was rescued by in vitro opsonization of A. fumigatus conidia and in vivo administration of murine SAP. Mouse and human SAP bound conidia, activated the complement cascade and increased phagocytosis by mouse and human neutrophils. Defective resistance of Apcs-/- SAP-deficient mice was associated with defective in vivo recruitment of neutrophils and phagocytosis in the lungs. The opsonic activity of SAP was dependent on the classical pathway of complement activation. In immunosuppressed mice, SAP administration protected hosts against A. fumigatus infection and death. In the context of a study of hematopoietic stem-cell transplantation, genetic variation in the human APCS gene was associated with susceptibility to invasive pulmonary aspergillosis. Thus, SAP is a fluid phase pattern recognition molecule essential for resistance against A. fumigatus. The results reported here also suggest that SAP has therapeutic potential against an infectious agent which represents a formidable clinical challenge.

2021-04-20 | GSE163533 | GEO

Response of A549 cells treated with Aspergillus fumigatus [WT-GC_vs_PrtT-GC]

Project description:Response of A549 cells treated with Aspergillus fumigatus wild type germinating conidia (WT_GC) or PrtT protease deficient mutant conidia (PrtT-GC) or inert acrylic 2-4 micron beads (Beads) for 8h Aspergillus fumigatus is the most commonly encountered mold pathogen of humans, predominantly infecting the respiratory system. Colonization and penetration of the lung alveolar epithelium is a key but poorly understood step in the infection process. This study focused on identifying the transcriptional and cell-signaling responses activated in A549 alveolar carcinoma cells incubated in the presence of A. fumigatus wild-type and ÎPrtT protease-deficient germinating conidia and culture filtrates (CF). Microarray analysis of exposed A549 cells identified distinct classes of genes whose expression is altered in the presence of germinating conidia and CF and suggested the involvement of both NFkB and MAPK signaling pathways in mediating the cellular response. Phosphoprotein analysis of A549 cells confirmed that JNK and ERK1/2 are phosphorylated in response to CF treatment in a protease-dependent manner. Inhibition of JNK or ERK1/2 kinase activity substantially decreased CF-induced cell damage, including cell peeling, actin-cytoskeleton damage, and reduction in metabolic activity and necrotic death. These results suggest that inhibition of MAPK-mediated host responses to treatment with A. fumigatus CF decreases cellular damage, a finding with possible clinical implications. 2 independent controls (uninfected A549 cells) as ctrl3_1-2, 2 independent treatments of A549 cells with wild-type A. fumigatus germinating conidia (WT_GC_1-2 ) , 2 independent treatments of A549 cells with inert acrylic beads (Beads 1-2), 3 independent treatments of A549 cells with A. fumigatus prtT mutant germinating conidia (PrtT-GC_1-3).

2010-10-29 | E-GEOD-24984 | biostudies-arrayexpress

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