Transcriptom profiling of in vitro induced exhauted CD8+ T cells
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ABSTRACT:
PROVIDER: PRJNA631227 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR11734909_1.fastq.gz | Fastqsanger.gz | |||
| SRR11734909_2.fastq.gz | Fastqsanger.gz | |||
| SRR11734910_1.fastq.gz | Fastqsanger.gz | |||
| SRR11734910_2.fastq.gz | Fastqsanger.gz | |||
| SRR11734911_1.fastq.gz | Fastqsanger.gz |
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Project description:By comparing transcriptome profiling of in vitro repeated stimulated cells with without stimulation and single peptide stimulated cells, to evaluated if the transcriptome profiling of repeated stimulated cells meet the features of exhausted cells. CD8+ T cells purified from OT-I mice were stimulated daily for five days with 10ng/ml OVA(257-264) peptide in the presence of IL-15 and IL-7. As controls, cells were either left without stimulation or stimulated only once with OVA(257-264) peptide for 48 hours and then washed and cultured without peptide for an additional 3 days. On day 5, cells were harvested. Live CD8+ T cells were sorted and RNAseq were performed after RNA isolation. It was found that the transcriptional prolife of the repeat peptide stimulated cells is clearly distinct to those of the single and non-stimulated cells. Among the upregulated genes, multiple inhibitory receptors encoding genes were presented on the top of this list. The genes for the markers of the terminal differentiated effector cells, like GzmB, GzmC and Prf1 were also significantly upregulated on the repeat peptide stimulated cells. The expression pattern of exhaustion specific transcription factors were identified in repeat stimulated cells. In vitro repeat peptide stimulation results in the transcriptional changes of in vivo exhausted CD8+ T cells from chronic viral infections which serve as the benchmark of CTL exhaustion.
2020-05-09 | GSE150120 | GEO
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Project description:Transcriptional profiling of mouse in-vitro derived CD8+ cells (obtained after culturing BM-HSCs isolated from B6 mice) with OP9-DL1 cells for 35 days were compared to CD8+ thymocytes isolated from 4-5 weeks old B6 mice. Our goal was to determine similarities and differences in gene expression profile between in vitro-derived CD8+ cells and CD8+ cells isolated from the thymus. The procedure of in vitro differentiation of HSC using OP9-DL1 cocultures was previously described by: Schmitt, T. M., and J. C. Zuniga-Pflucker. 2002. Induction of T cell development from hematopoietic progenitor cells by delta-like-1 in vitro. Immunity 17:749-756 and Holmes, R., and J. C. Zuniga-Pflucker. 2009. The OP9-DL1 system: generation of T-lymphocytes from embryonic or hematopoietic stem cells in vitro. Cold Spring Harb Protoc 2009:pdb prot5156. Biological replicates: 2 different samples of in vitro-derived CD8+ T cells from day 35 were compared to 2 different samples of thymic CD8+ cells.
2012-09-05 | E-GEOD-37669 | biostudies-arrayexpress
Project description:Background: Infrared A radiation (IRA 760-1440 nm) is a major component of solar radiation and, similar to ultraviolet (UV) radiation, causes photoaging of human skin by increasing the expression of matrixmetalloproteinase-1 (MMP-1) expression in human skin fibroblasts. This gene-regulatory effect resulted from the IRA induced activation of the pleiotropic MAPKinase ERK1/2 signaling pathway, indicating that the IRA response extends beyond MMP-1. In the present study we have therefore assessed the IRA-induced transcriptom in primary human skin fibroblasts. Results: Microarray analysis revealed 599 transcripts to be regulated by physiologically relevant doses of IRA in primary human skin fibroblasts. The IRA induced transcriptom differed from changes known to be induced by UVB or UVA radiation in the same cell type. IRA responsive genes include four categories: extracellular matrix, calcium homeostasis, stress signaling, and apoptosis. These results were confirmed by realtime PCR experiments analyzing thirteen of these genes representing these four categories. By means of chemical inhibitors of known signaling pathways we next show that besides ERK1/2, the p38-, JNK-, PI3K/AKT-, STAT3-, and IL-6 as well as calcium mediated signaling pathways are functionally involved in the IRA gene response and that a major part of it is triggered by mitochondrial, and to a lesser extent non-mitochondrial production of reactive oxygen species. Conclusion: This study identifies IRA radiation as a potent modulator of gene expression in human skin cells. The IRA response is specific and involves genes which are of critical importance for the homeostasis of human skin. Our data confirm the previous notion that IRA contributes to premature skin aging and indicate that further biological effects may result from IRA exposure of human skin. To identify genes differentially regulated after IRA irradiation we analyzed a set of nine independent samples pairs (IRA irradiated vs. sham irradiated), using human primary human dermal fibroblasts from three different donors (termed F1 to F3). Cells between passage number five to ten were exposed in vitro to a dose of 860 J/cm2 IRA. IRA irradiation lasted 40 min leading to a total dose of 860 J/cm2. The culture dishes were placed on a cooled plate connected to thermostated bath (Thermo Haake GmbH, Karlsruhe, Germany) to maintain temperatures below 37°C during IRA irradiation. For irradiation, medium was replaced by phosphate-buffered saline (37°C, Gibco). Control cells (sham) were held on a 37°C thermostated plate under similar conditions without irradiation. Following the treatment cells were cultivated for 24 hours with serum free MEM culture medium at 37°C.
2010-07-10 | E-GEOD-17046 | biostudies-arrayexpress
Project description:Analyzing and comparing proteome and phosphoproteome profiling between CD8+ DC and CD8- DC not only will be very helpful to understand the molecular mechanisms regulating CD8+/- DC differentiation and function but also can screen CD8+ or CD8- DC specific regulatory pathways or molecules.
2018-06-11 | PXD006875 | Pride
Project description:Transcriptional profiling of mouse in-vitro derived CD8+ cells (obtained after culturing BM-HSCs isolated from B6 mice) with OP9-DL1 cells for 35 days were compared to CD8+ thymocytes isolated from 4-5 weeks old B6 mice. Our goal was to determine similarities and differences in gene expression profile between in vitro-derived CD8+ cells and CD8+ cells isolated from the thymus. The procedure of in vitro differentiation of HSC using OP9-DL1 cocultures was previously described by: Schmitt, T. M., and J. C. Zuniga-Pflucker. 2002. Induction of T cell development from hematopoietic progenitor cells by delta-like-1 in vitro. Immunity 17:749-756 and Holmes, R., and J. C. Zuniga-Pflucker. 2009. The OP9-DL1 system: generation of T-lymphocytes from embryonic or hematopoietic stem cells in vitro. Cold Spring Harb Protoc 2009:pdb prot5156.
2012-09-05 | GSE37669 | GEO
Project description:Background: Infrared A radiation (IRA 760-1440 nm) is a major component of solar radiation and, similar to ultraviolet (UV) radiation, causes photoaging of human skin by increasing the expression of matrixmetalloproteinase-1 (MMP-1) expression in human skin fibroblasts. This gene-regulatory effect resulted from the IRA induced activation of the pleiotropic MAPKinase ERK1/2 signaling pathway, indicating that the IRA response extends beyond MMP-1. In the present study we have therefore assessed the IRA-induced transcriptom in primary human skin fibroblasts. Results: Microarray analysis revealed 599 transcripts to be regulated by physiologically relevant doses of IRA in primary human skin fibroblasts. The IRA induced transcriptom differed from changes known to be induced by UVB or UVA radiation in the same cell type. IRA responsive genes include four categories: extracellular matrix, calcium homeostasis, stress signaling, and apoptosis. These results were confirmed by realtime PCR experiments analyzing thirteen of these genes representing these four categories. By means of chemical inhibitors of known signaling pathways we next show that besides ERK1/2, the p38-, JNK-, PI3K/AKT-, STAT3-, and IL-6 as well as calcium mediated signaling pathways are functionally involved in the IRA gene response and that a major part of it is triggered by mitochondrial, and to a lesser extent non-mitochondrial production of reactive oxygen species. Conclusion: This study identifies IRA radiation as a potent modulator of gene expression in human skin cells. The IRA response is specific and involves genes which are of critical importance for the homeostasis of human skin. Our data confirm the previous notion that IRA contributes to premature skin aging and indicate that further biological effects may result from IRA exposure of human skin.
2010-07-10 | GSE17046 | GEO
Project description:CD8+ NKT cells are naturally occurring but rare T cells that express both T cell and natural killer (NK) cell markers. These cells may play key roles in establishing tolerance to self antigens; however, the mechanism of action and the molecular profiles of these cells are poorly characterized due to their extremely low frequencies. We developed a highly efficient in vitro conversion/expansion protocol for such cells and extensively characterized their functional and molecular phenotypes using a variety of genomic and immunological techniques. We used microarrays to elucidate the molecular mechanisms underlying the suppression function of the converted CD8+ NKT cells. Keywords: Cell type comparison To identify candidate genes that might be related to the suppressive activity, the CD8+ NKT cells with potent suppressive activity were compared to three different control cells: 1) CD4+ T cells (with very weak suppression) cultured under the same condition as a control for culture condition, 2) naïve CD8 T cells as the baseline expression level, and 3) CD8+ T cells activated using conventional activation protocols. The last two cell populations do not possess suppressive activity. This data set allows us to examine a number of different questions
2009-02-01 | E-GEOD-10813 | biostudies-arrayexpress
Project description:Purified primary human CD8 T cells were stimulated in vitro in parallel using a magnetic bead conjugated to either anti-CD3/28 or anti-CD2/3/28 antibodies. Cells were cultured for 6 days in complete RPMI1640 and 10ng/ml IL2 and were harvested on day 6 and RNA was isolated for further labelling and hybridisation.
2015-08-12 | E-MTAB-3470 | biostudies-arrayexpress