13033
2013
eng
e50695
2
8
article
1
2016-03-18
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Accumulation of Splice Variants and Transcripts in Response to PI3K Inhibition in T Cells
Background
Measles virus (MV) causes T cell suppression by interference with phosphatidylinositol-3-kinase (PI3K) activation. We previously found that this interference affected the activity of splice regulatory proteins and a T cell inhibitory protein isoform was produced from an alternatively spliced pre-mRNA.
Hypothesis
Differentially regulated and alternatively splice variant transcripts accumulating in response to PI3K abrogation in T cells potentially encode proteins involved in T cell silencing.
Methods
To test this hypothesis at the cellular level, we performed a Human Exon 1.0 ST Array on RNAs isolated from T cells stimulated only or stimulated after PI3K inhibition. We developed a simple algorithm based on a splicing index to detect genes that undergo alternative splicing (AS) or are differentially regulated (RG) upon T cell suppression.
Results
Applying our algorithm to the data, 9% of the genes were assigned as AS, while only 3% were attributed to RG. Though there are overlaps, AS and RG genes differed with regard to functional regulation, and were found to be enriched in different functional groups. AS genes targeted extracellular matrix (ECM)-receptor interaction and focal adhesion pathways, while RG genes were mainly enriched in cytokine-receptor interaction and Jak-STAT. When combined, AS/RG dependent alterations targeted pathways essential for T cell receptor signaling, cytoskeletal dynamics and cell cycle entry.
Conclusions
PI3K abrogation interferes with key T cell activation processes through both differential expression and alternative splicing, which together actively contribute to T cell suppression.
PLoS ONE
10.1371/journal.pone.0050695
urn:nbn:de:bvb:20-opus-130335
PLoS ONE 8(2): e50695. doi:10.1371/journal.pone.0050695
Alice Riedel
Boitumelo Mofolo
Elita Avota
Sibylle Schneider-Schaulies
Ayton Meintjes
Nicola Mulder
Susanne Kneitz
eng
uncontrolled
T cells
eng
uncontrolled
gene regulation
eng
uncontrolled
alternative splicing
eng
uncontrolled
measles virus
eng
uncontrolled
T cell receptors
eng
uncontrolled
reverse transcriptase-polymerase chain reaction
eng
uncontrolled
cell cycle and cell division
eng
uncontrolled
TCR signaling cascade
Biowissenschaften; Biologie
open_access
Institut für Virologie und Immunbiologie
Theodor-Boveri-Institut für Biowissenschaften
Förderzeitraum 2012
Universität Würzburg
11289
2014
eng
article
1
2015-05-08
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Gene Expression Profiles of Human Dendritic Cells Interacting with Aspergillus fumigatus in a Bilayer Model of the Alveolar Epithelium/Endothelium Interface
The initial stages of the interaction between the host and Aspergillus fumigatus at the alveolar surface of the human lung are critical in the establishment of aspergillosis. Using an in vitro bilayer model of the alveolus, including both the epithelium (human lung adenocarcinoma epithelial cell line, A549) and endothelium (human pulmonary artery epithelial cells, HPAEC) on transwell membranes, it was possible to closely replicate the in vivo conditions. Two distinct sub-groups of dendritic cells (DC), monocyte-derived DC (moDC) and myeloid DC (mDC), were included in the model to examine immune responses to fungal infection at the alveolar surface. RNA in high quantity and quality was extracted from the cell layers on the transwell membrane to allow gene expression analysis using tailored custom-made microarrays, containing probes for 117 immune-relevant genes. This microarray data indicated minimal induction of immune gene expression in A549 alveolar epithelial cells in response to germ tubes of A. fumigatus. In contrast, the addition of DC to the system greatly increased the number of differentially expressed immune genes. moDC exhibited increased expression of genes including CLEC7A, CD209 and CCL18 in the absence of A. fumigatus compared to mDC. In the presence of A. fumigatus, both DC subgroups exhibited up-regulation of genes identified in previous studies as being associated with the exposure of DC to A. fumigatus and exhibiting chemotactic properties for neutrophils, including CXCL2, CXCL5, CCL20, and IL1B. This model closely approximated the human alveolus allowing for an analysis of the host pathogen interface that complements existing animal models of IA.
10.1371/journal.pone.0098279
urn:nbn:de:bvb:20-opus-112893
PLoS ONE 9(5): e98279. doi:10.1371/journal.pone.0098279
Charles Oliver Morton
Mirjam Fliesser
Marcus Dittrich
Tobias Müller
Ruth Bauer
Susanne Kneitz
William Hope
Thomas Richard Rogers
Hermann Einsele
Jürgen Löffler
eng
uncontrolled
aspergillus fumigatus
eng
uncontrolled
gene expression
eng
uncontrolled
immune receptors
eng
uncontrolled
immune response
eng
uncontrolled
denritic cells
eng
uncontrolled
B cell receptors
eng
uncontrolled
gene regulation
eng
uncontrolled
RNA extraction
Medizin und Gesundheit
open_access
Medizinische Klinik und Poliklinik II
Theodor-Boveri-Institut für Biowissenschaften
Förderzeitraum 2014
Universität Würzburg
https://opus.bibliothek.uni-wuerzburg.de/files/11289/097_Löffler_PLoS.pdf