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The temporal dynamics of differential gene expression in Aspergillus fumigatus interacting with human immature dendritic cells in vitro.

Morton CO, Varga JJ, Hornbach A, Mezger M, Sennefelder H, Kneitz S, Kurzai O, Krappmann S, Einsele H, Nierman WC, Rogers TR, Loeffler J - PLoS ONE (2011)

Bottom Line: RNA was extracted from both organisms at each time point and hybridised to microarrays. iDC cell death increased at 6 h in the presence of A. fumigatus which coincided with fungal germ tube emergence; >80% of conidia were associated with iDC.Over the time course A. fumigatus differentially regulated 210 genes, FunCat analysis indicated significant up-regulation of genes involved in fermentation, drug transport, pathogenesis and response to oxidative stress.This study shows that A. fumigatus adapts to phagocytosis by iDCs by utilising genes that allow it to survive the interaction rather than just up-regulation of specific virulence genes.

View Article: PubMed Central - PubMed

Affiliation: Department of Clinical Microbiology, Trinity College Dublin, Dublin, Ireland.

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. Monocyte-derived immature DCs (iDC) were infected with viable resting conidia of Aspergillus fumigatus (Af293) for 12 hours at an MOI of 5; cells were sampled every three hours. RNA was extracted from both organisms at each time point and hybridised to microarrays. iDC cell death increased at 6 h in the presence of A. fumigatus which coincided with fungal germ tube emergence; >80% of conidia were associated with iDC. Over the time course A. fumigatus differentially regulated 210 genes, FunCat analysis indicated significant up-regulation of genes involved in fermentation, drug transport, pathogenesis and response to oxidative stress. Genes related to cytotoxicity were differentially regulated but the gliotoxin biosynthesis genes were down regulated over the time course, while Aspf1 was up-regulated at 9 h and 12 h. There was an up-regulation of genes in the subtelomeric regions of the genome as the interaction progressed. The genes up-regulated by iDC in the presence of A. fumigatus indicated that they were producing a pro-inflammatory response which was consistent with previous transcriptome studies of iDC interacting with A. fumigatus germ tubes. This study shows that A. fumigatus adapts to phagocytosis by iDCs by utilising genes that allow it to survive the interaction rather than just up-regulation of specific virulence genes.

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RT-qPCR validation of differential gene expression observed in microarray experiments.(a) Gene expression data from the A. fumigatus microarray shown as mean log2 values of ratios of relative expression determined by RT-qPCR compared to array data. (b) Gene expression data from the immune gene microarray shown as mean log2 values of ratios of relative expression determined by RT-qPCR compared to array data. Error bars in both charts indicate the standard error.
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pone-0016016-g005: RT-qPCR validation of differential gene expression observed in microarray experiments.(a) Gene expression data from the A. fumigatus microarray shown as mean log2 values of ratios of relative expression determined by RT-qPCR compared to array data. (b) Gene expression data from the immune gene microarray shown as mean log2 values of ratios of relative expression determined by RT-qPCR compared to array data. Error bars in both charts indicate the standard error.

Mentions: PCR results confirmed the differential expression of selected genes from the micro-array analyses (Fig. 5a and 5b); genes tested from both arrays show the same expression patterns for both qRT-PCR and microarray. These genes were selected from a list of genes that showed the greatest differential expression on the array.


The temporal dynamics of differential gene expression in Aspergillus fumigatus interacting with human immature dendritic cells in vitro.

Morton CO, Varga JJ, Hornbach A, Mezger M, Sennefelder H, Kneitz S, Kurzai O, Krappmann S, Einsele H, Nierman WC, Rogers TR, Loeffler J - PLoS ONE (2011)

RT-qPCR validation of differential gene expression observed in microarray experiments.(a) Gene expression data from the A. fumigatus microarray shown as mean log2 values of ratios of relative expression determined by RT-qPCR compared to array data. (b) Gene expression data from the immune gene microarray shown as mean log2 values of ratios of relative expression determined by RT-qPCR compared to array data. Error bars in both charts indicate the standard error.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3021540&req=5

pone-0016016-g005: RT-qPCR validation of differential gene expression observed in microarray experiments.(a) Gene expression data from the A. fumigatus microarray shown as mean log2 values of ratios of relative expression determined by RT-qPCR compared to array data. (b) Gene expression data from the immune gene microarray shown as mean log2 values of ratios of relative expression determined by RT-qPCR compared to array data. Error bars in both charts indicate the standard error.
Mentions: PCR results confirmed the differential expression of selected genes from the micro-array analyses (Fig. 5a and 5b); genes tested from both arrays show the same expression patterns for both qRT-PCR and microarray. These genes were selected from a list of genes that showed the greatest differential expression on the array.

Bottom Line: RNA was extracted from both organisms at each time point and hybridised to microarrays. iDC cell death increased at 6 h in the presence of A. fumigatus which coincided with fungal germ tube emergence; >80% of conidia were associated with iDC.Over the time course A. fumigatus differentially regulated 210 genes, FunCat analysis indicated significant up-regulation of genes involved in fermentation, drug transport, pathogenesis and response to oxidative stress.This study shows that A. fumigatus adapts to phagocytosis by iDCs by utilising genes that allow it to survive the interaction rather than just up-regulation of specific virulence genes.

View Article: PubMed Central - PubMed

Affiliation: Department of Clinical Microbiology, Trinity College Dublin, Dublin, Ireland.

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. Monocyte-derived immature DCs (iDC) were infected with viable resting conidia of Aspergillus fumigatus (Af293) for 12 hours at an MOI of 5; cells were sampled every three hours. RNA was extracted from both organisms at each time point and hybridised to microarrays. iDC cell death increased at 6 h in the presence of A. fumigatus which coincided with fungal germ tube emergence; >80% of conidia were associated with iDC. Over the time course A. fumigatus differentially regulated 210 genes, FunCat analysis indicated significant up-regulation of genes involved in fermentation, drug transport, pathogenesis and response to oxidative stress. Genes related to cytotoxicity were differentially regulated but the gliotoxin biosynthesis genes were down regulated over the time course, while Aspf1 was up-regulated at 9 h and 12 h. There was an up-regulation of genes in the subtelomeric regions of the genome as the interaction progressed. The genes up-regulated by iDC in the presence of A. fumigatus indicated that they were producing a pro-inflammatory response which was consistent with previous transcriptome studies of iDC interacting with A. fumigatus germ tubes. This study shows that A. fumigatus adapts to phagocytosis by iDCs by utilising genes that allow it to survive the interaction rather than just up-regulation of specific virulence genes.

Show MeSH
Related in: MedlinePlus