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Differential expression of miRNA in control and CD3/CD28mAb-treated Jurkat cells

ABSTRACT: The cultured cell line Jurkat is frequently employed in studies of T cell function. Here we identified the microRNAs expressed in Jurkat cells in the presence and absence of CD3/CD28mAb treatment.

ORGANISM(S): Homo sapiens

PROVIDER: GSE23430 | GEO | 2010/12/31

SECONDARY ACCESSION(S): PRJNA131097

REPOSITORIES: GEO

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Differential expression of miRNA in control and CD3/CD28mAb-treated Jurkat cells

Project description:The cultured cell line Jurkat is frequently employed in studies of T cell function. Here we identified the microRNAs expressed in Jurkat cells in the presence and absence of CD3/CD28mAb treatment. Analyzed the expression of microRNAs extracted from untreated Jurkat cells as a control and Jurkat cells treated with CD3/CD28mAb.

2010-12-31 | E-GEOD-23430 | biostudies-arrayexpress

CD3/CD28 signaling, Jurkat E6.1, time course 27 mins

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Profiling of Jurkat T cells activated with CD3, CD28 and PMA at 1 and 8 hours

Project description:T lymphocytes are orchestrators of adaptive immunity. Naïve T cells may differentiate into the Th1, Th2, Th17 or iTreg phenotype, depending on environmental co-stimulatory signals. In order to identify the genes and pathways involved in differentiation of Jurkat T cells towards Th1 and Th2 subtypes we performed comprehensive transcriptome analyses of Jurkat T cells stimulated with various stimuli an pathway inhibitors Jurkat T cells were treated with CD3/CD28 and CD3/PMA and CD28/PMA. RNA was isolated after 1 and 8 hrs stumalation.

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Project description:T lymphocytes are orchestrators of adaptive immunity. Naïve T cells may differentiate into the Th1, Th2, Th17 or iTreg phenotype, depending on environmental co-stimulatory signals. In order to identify the genes and pathways involved in differentiation of Jurkat T cells towards Th1 and Th2 subtypes we performed comprehensive transcriptome analyses of Jurkat T cells stimulated with various stimuli an pathway inhibitors Jurkat T cells were treated with CD3, CD28 and PMA and all pairwise combinations, in the presence of DMSO (control) or kinase inhibitors. RNA was isolated after 8 hrs incubation.

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Phosphoproteomics quantitative characterization in Jurkat E6.1 and Jurkat Gamma1.

Project description:Phosphorylation is a reversible post-translational modification, playing a vital role in protein function. In T cells, protein phosphorylation is the key mechanism regulating T cell receptor – driven signaling pathways. In order to gain insights into the phosphoproteome evolution of T cell activation, we performed a large-scale quantitative phosphoproteomics study of Jurkat E6.1 (wild type) and Jurkat gamma1 (Phospholipase gamma1 null) cell clones upon costimulation with anti-CD3 and anti-CD28 antibodies at times ranging from 15 min to as long as 120 min. In total, we identified 5585 phosphopeptides belonging to 2008 phosphoproteins from both cell clones. We detected 130 and 114 novel phosphopeptides in Jurkat E6.1 and Jurkat gamma1 clones, respectively. A significantly lower number of proteins containing regulated phosphorylation sites were identified in Jurkat gamma1 in comparison to Jurkat E6.1, reflecting the vital role of Phospholipase gamma1 in T cell signaling. Several new phosphorylation sites from lymphocyte-specific protein tyrosine kinase (LCK) were identified. Of these, Serine-121 showed significant changes in JE6.1 while only small changes in the Jgamma1 clone.

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Profiling of Jurkat T cells activated with CD3, CD28 and PMA and multiple kinase inhibitors

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Project description:Jurkat clone E6-1 is a human T-cell line derived from a paediatric case of acute leukaemia, and is frequently employed as a model T-cell for immunology research. In this study, we characterised genomic, transcriptomic, and functional features of Jurkat clone E6-1 cells stored in three different laboratories, and compared these to Jurkat E6-1 cells from the ATCC. Jurkat clone E6-1 populations were first karyotyped, which results showing marked cytogenomic heterogeneity between cell line. These results were confirmed through chromosomal microarray analysis. Whole exome sequencing was then conducted on each cell line, and revealed distinct mutational profiles between populations. Mutational differences between Jurkat clone E6-1 populations detected by cytogenomic approaches and whole exome sequencing led to divergent transcriptomic profiles identified by bulk RNA sequencing, which, in turn, impacted cellular phenotype and function shown by flow cytometry and multiplex cytokine bead array data. Overall, we found substantial genomic heterogeneity between and within Jurkat clone E6-1 cells, resulting in significant differences in gene expression and function. This research highlights the extreme genomic instability of Jurkat clone E6-1 cells, the importance of monitoring cell passage number and procuring fresh cell line stocks, and the value of multi-omic approaches to characterise cell lines to improve the accuracy, reproducibility, and translatability of research.

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miRNA profiling of Jurkat with stably integrated HTLV1-LTR in the presence of HTLV-1 Tax

Project description:RNA interference (RNAi) is a natural cellular mechanism to silence gene expression and is predominantly mediated by microRNAs (miRNAs) that target messenger RNA. Viruses can manipulate the cellular processes necessary for their replication by targeting the host RNAi machinery. This study explores the effect of human T-cell leukemia virus type 1 (HTLV-1) transactivating protein Tax on the RNAi pathway in the context of a chromosomally integrated viral long terminal repeat (LTR) using a CD4(+) T-cell line, Jurkat. Jurkat cells with stably integrated HTLV1-LTR were transfected with either pCMV-Tax or pCMV. Total miRNA was isolated, labeled and hybridized to the miRNA microarray as described by the manufacturer (Invitrogen). The hybridized slides were then analyzed on a GenePix scanner.

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