Project description:Gene expression profiles of human immature dendritic cells after 3h, 6h, 9h and 12h of co-cultivation with Aspergillus fumigatus were compared to expression profiles from human immature dendritic cells after 3h, 6h, 9h and 12h of cultivation. We used microarrays to detail the gene expression of human immature dendritic cells after 3h, 6h, 9h and 12h of co-cultivation with Aspergillus fumigatus
Project description:Gene expression profiles of human immature dendritic cells after 3h, 6h, 9h and 12h of co-cultivation with Aspergillus fumigatus were compared to expression profiles from human immature dendritic cells after 3h, 6h, 9h and 12h of cultivation.
Project description:This SuperSeries is composed of the following subset Series: GSE21353: Gene expression profiles of human immature dendritic cells after 3h, 6h and 12h of co-cultivation with Aspergillus fumigatus GSE28806: The temporal dynamics of differential gene expression in human alveolar epithelial and endothelial cells interacting with the human pathogenic mould Aspergillus fumigatus in vitro Refer to individual Series
Project description:In a whole-transcriptome study, cellular responses of DCs confronted with the fungi A. fumigatus, C. albicans or the bacterial cell wall component LPS were investigated. Therefore DCs of four independent donors were harvested after 6 and 12 hours co-culture with A. fumigatus, C. albicans and LPS and analyzed with Affymetrix whole genome expression arrays. In general, transcriptomic analysis revealed a clustering of the A. fumigatus and C. albicans stimulated DCs. However, LPS and fungi-dependent gene expression showed more common similarities compared to the untreated control. Stimulation with LPS induced a differential regulation of 2793 and 2863 genes after 6 /12h, while confrontation with A. fumigatus and C. albicans resulted in 743/1076 and 1729/974 differentially regulated genes, respectively. Kruppel-like factor 4 (KLF4) was identified as the only transcription factor that was down-regulated in DCs by both fungi but induced by stimulation with LPS. Human DCs were generated from 4 independent, healthy blood donors. moDCs were either left untreated or co-cultivated with Aspergillus fumigatus 46645 germ tubes or Candida albicans SC5314 (MOI1) and or LPS (1µg/ml) for 6h.
Project description:Within the last decades, invasive fungal infections have gained increasing significance. They are characterized by high mortality rates and are often caused Candida albicans and Aspergillus fumigatus. The increasing number of infections underlines the necessity for additional anti-fungal therapies, which require an extended knowledge of gene regulations during fungal infection. MicroRNAs are regulators of important cellular processes, including immune response. By analyzing their regulation and impact on target genes, novel therapeutic approaches may be developed. Here, we examine the role of microRNAs in human dendritic cells during fungal infections. Dendritic cells represent the bridge between the innate and the adaptive immune systems. Therefore, analysis of gene regulation of dendritic cells is of particular significance. By applying next-generation sequencing of small RNAs, we quantify microRNA expression in monocyte-derived dendritic cells after 6 and 12h of infection with C. albicans and A. fumigatus as well as treatment with LPS. We use two different tools and an online database to determine potential target genes. We identified 29 microRNAs that are differentially regulated after infection by the fungi or LPS. Two and five of them are specific for fungal infections after 6h and 12h, respectively. We further validated interactions of miR-132-5p and miR-212-5p with immunological relevant target genes, such as FKBP1B, KLF4, and SPN, on both RNA and protein level. Our results indicate a fine-tuning function of these microRNAs during fungal infections. Beyond that, we identified previously undiscovered microRNAs. We validated three novel microRNAs via qRT-PCR. A comparison with known microRNAs revealed possible relations with the miR-378 family and miR-1260a/b for two of them, while the third one features a unique sequence with no resemblance to known microRNAs. In summary, this study analyzes the effect of known microRNAs in dendritic cells during fungal infections and proposes novel microRNAs that could be experimentally verified.
Project description:Gene expression profiles of human immature dendritic cells co-cultivated with Aspergillus fumigatus, Candida albicans and LPS for 6h
Project description:Dendritic cells from three healthy human donors were cultured in the presence or absence of either Aspergillus fumigatus, Saccharomyces cerevisiae, Candida albicans or Candida parapsilosis. Each of the four fungi were also cultured in the absence of human cells. RNA-sequencing was used to evaluate differences in the transcriptomes of human cells challenged and unchallenged with each fungal pathogen, as well as in those of each fungus challenged and unchallenged by cells from the human immune system.
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. We used microarrays to detail the gene expression of human alveloar epithelial and endothelials cells after 3h and 6h of co-cultivation with Aspergillus fumigatus and monocyte-derived or myeloid dendritic cells
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
