MiRNA profilng in IL-2 induced CD4+T cells
Ontology highlight
ABSTRACT:
ORGANISM(S): Homo sapiens
PROVIDER: GSE68609 | GEO | 2015/05/07
SECONDARY ACCESSION(S): PRJNA283225
REPOSITORIES: GEO
Similar Datasets
Project description:To evaluate role of miRNAs in proliferation and death of T cell, we performed miRNA profiling in activated CD4+T cells after IL-2 induction and depletion. Proliferation rate of IL-2 induced cells was measured by MTT assay. Then quantitative RT-PCR arrays on 739 miRNAs revealed up- and down-regulation of 170 miRNAs in IL-2 induced CD4+T cells relative to IL-2 depleted ones. For miRNA profiling, total RNA from activated CD4+T cells in the presence and the absence of IL-2 (0.75 n/ml) after 3 days was extracted. RNA isolation was carried out using miRCURYTM RNA isolation kit. cDNA synthesis and Q-RT-PCR were performed with miRCURY LNATM universal RT microRNA PCR kit. Both kits were purchased from Exiqon (Vedbaek, Denmark). According to manufacturer’s instructions, to synthesize cDNA, total RNA was added to enzyme mix, reaction buffer, and RNA spike-in (control RNA). Then, the mixture was incubated 60 min at 42 °C and 5 min at 95 °C. To perform Q-RT-PCR array, cDNA was added to SYBR green master mix and was dispensed into the 384-well plates (by a specific LNATM primer pair/ well); then the reaction was performed using a real-time PCR cycler (Roche LC480, USA) according to this program: 95°C for 10 min, 45 cycles at 95˚C for 10 sec, 60˚C for 1 min. For miRNA profiling, two 384-well plates (human panel I & II) were used and each reaction was performed twice. Each panel contained 6 reference primer sets as well as one control primer set (that targeted a synthetic spike-in RNA) and an inter-plate calibrator in triplicate. To analyze Q-RT-PCR data, GenEx software and ΔΔCT method were used. By Normfinder, SNORD49A (snoRNA U49) gene was identified as optimal reference gene for normalization. Fold change of each miRNA was calculated by the equation 2−ΔΔCT and logarithmically transformed (base 2). P-values were adjusted for multiple comparisons using Dunn-Benferroni correction.
2015-05-07 | E-GEOD-68609 | biostudies-arrayexpress
Project description:Homeostatic proliferation, driven by cytokines including IL-7, maintains stable T cell populations over time and expands these during lymphopenia, for example following stem cell transplantation. Yet, impaired T cell expansion in this context is associated with diminished immune protection, whilst excessive proliferation contributes to graft versus host disease. This highlights the need to understand molecular mechanisms regulating this process. Here, we studied the metabolic basis for IL-7-driven CD4+ T cell proliferation. We observed that IL-7 promotes glucose uptake and metabolism, including usage for nucleotide synthesis and oxidation in the tricarboxylic acid (TCA) cycle. Of note, contrary to other TCA metabolites, glucose-derived citrate does not accumulate in IL-7 treated cells, indicating diversion into other processes. In agreement, IL-7 promotes glucose-dependent histone acetylation and chromatin accessibility during proliferation. This is particularly notable at gene loci of the amino acid-sensing Ragulator complex. Consistently, expression of its subunit LAMTOR5 is promoted by IL-7 in a glucose-dependent manner. Functionally, glucose availability determined amino acid-dependent mTOR activation in CD4+ T cells, confirming integrated sensing of these nutrients. LAMTOR5 deletion impaired IL-7-mediated CD4+ T cell expansion, establishing that glycolysis in absence of Ragulator activation was insufficient to initiate this process. Clinically, CD4+ T cells isolated from patients following myeloablative autologous stem cell transplantation demonstrate coordinated upregulation of genes encoding glycolytic and TCA cycle enzymes, as well as amino acid sensing machinery and mTOR targets. Our findings uncover integrated sensing of glucose and amino acid metabolism and demonstrate its relevance during homeostatic T cell proliferation.
2024-11-27 | MSV000096546 | MassIVE
Project description:CD4+ T cells orchestrate both humoral and cytotoxic immune responses. While it is known that CD4+ T cell proliferation relies on autophagy, direct identification of the autophagosomal cargo involved is still missing. Here, we created a transgenic mouse model, which, for the first time, enables us to directly map the proteinaceous content of autophagosomes in any primary cell by LC3 proximity labelling. IL-7Rα, a cytokine receptor mostly found in naïve and memory T cells, was reproducibly detected in autophagosomes of activated CD4+ T cells. Consistently, CD4+ T cells lacking autophagy showed increased IL-7Rα surface expression, while no defect in internalisation was observed. Mechanistically, excessive surface IL-7Rα sequestrates the common gamma chain, impairing the IL-2R assembly and downstream signalling crucial for T cell proliferation. This study provides proof-of-principle that key autophagy substrates can be reliably identified with this model to help mechanistically unravel autophagy’s contribution to healthy physiology and disease.
2022-07-08 | PXD033733 | Pride
Project description:Chronic Lymphocytic Leukemia (CLL) cells multiply in secondary lymphoid tissue but the mechanisms leading to their proliferation are still uncertain. In addition to BCR-triggered signals, other microenvironmental factors might well be involved. In proliferation centres, leukemic B cells are in close contact with CD4+CD40L+ T cells. Therefore, we here dissected the signals provided by autologous activated T cells (Tact) to CLL cells. Although the gene expression profile induced by Tact was highly similar to that induced by sole CD40 signaling, an obvious difference was that Tact induced proliferation of CLL cells. We determined that stimulation with only CD40L+IL-21 was sufficient to induce robust proliferation in CLL cells. We then defined an IL-21-induced gene signature in CLL, containing components of JAK-STAT and apoptosis pathways, and this signature could be detected in lymph node (LN) samples from patients. Finally, we could detect IL-21 RNA and protein in LN, and IL-21 production ex vivo by LN CD4+CXCR5+ follicular helper T cells. These results indicate that, in addition to BCR signaling, activated T cells might contribute to CLL cell proliferation via CD40 and IL-21. Targeting these signaling pathways might offer new venues for treatment of CLL. CLL cells were cultured under different conditions for 16 hours and then sorted to purity as CD20+ CD5+ cells.
2013-10-07 | E-GEOD-50572 | biostudies-arrayexpress
Project description:Th17 cells are enriched by sorting FR4-CD4+ T cells from SKG mice. A large number of Th17 cells also develop spontaneously when CD4+ T cells from IFN-g-deficient (IFN-g-/-) BALB/c mice are transferred to T cell-deficient RAG2-deficient (RAG2-/-) mice and subjected to homeostatic proliferation, whereas they fail to develop in similar transfer of IL-6-deficient (IL-6-/-) CD4+ T cells to IL-6-/- RAG2-/- mice. To explore the functional molecules specifically expressed by Th17 cells, we conducted Gene Microarray analysis between 10-month-old SKG FR4-CD4+ cells and age-matched BALB/c FR4-CD4+ cells, and between IFN-g-/- CD4+ cells transferred to RAG2-/- mice and IL-6-/- CD4+ T cells transferred to IL-6-/- RAG2-/- mice. The analysis revealed that 1,556 and 115 genes were up-regulated in 10-month-old SKG FR4-CD4+ and IFN-g-/- CD4+ T cells after homeostatic proliferation, respectively, with 29 genes shared by the two groups of genes. The 29 genes included those encoding cytokines, chemokines, and their receptors, such as IL-1 receptor type1 (IL-1R1), IL-17, IL-22, IL-21, CCR6, and CCL20. Experiment Overall Design: We conducted Gene Microarray analysis between 10-month-old SKG FR4-CD4+ cells and age-matched BALB/c FR4-CD4+ cells, and between IFN-g-/- CD4+ cells transferred to RAG2-/- mice and IL-6-/- CD4+ T cells transferred to IL-6-/- RAG2-/- mice using GeneChip Mouse Genome 430 2.0 Array (Affymetrix). Each gruop has three replicates.
2008-06-16 | E-GEOD-9316 | biostudies-arrayexpress
Project description:Metabolites are thought as the end products in cellular regulatory processes and their levels show the strongest relationships with the phenotype. Previously, we showed that the administration of Clostridium butyricum MIYAIRI 588 (CBM 588) upregulated protectin D1, an anti-inflammatory lipid metabolite, in colon tissue under antibiotic therapy. However, how CBM 588 induces protectin D1 expression and whether the metabolite has anti-inflammatory effects on antibiotic-induced inflammation are unclear. Therefore, here, we evaluated the effect of CBM 588 on lipid metabolism and protectin D1 in gut protection from antibiotic-induced intestinal disorders. In the CBM 588 treatment group, expression levels of genes encoding lipid receptors related to the conversion of DHA to protectin D1, such as polyunsaturated fatty acid (PUFA) receptors, G-protein coupled receptor 120 (GPR120), and 15-lipoxygenase (LOX), were increased in colon tissue. CD4+ cells producing interleukin (IL)-4, the main component of T helper type 2 (Th2) cells that can activate 15-LOX, also increased in CBM 588-treated groups even after clindamycin co-administration. In addition, similar to CBM 588, exogenously administered protectin D1 reduced inflammatory cytokines, while IL-10 and TGF-β1, works as anti-inflammatory cytokines, were increased. Our data revealed that CBM 588 activated 15-LOX to enhance protectin D1 production by increasing IL-4-producing CD4+ cell population in the intestinal tract. Additionally, CBM 588-induced protectin D1 clearly upregulated IL-10-producing CD4+ cells to control antibiotic-induced gut inflammation. We provide new insights into CBM 588-mediated lipid metabolism induction for the treatment of gut inflammatory diseases.
2020-10-14 | MTBLS2087 | MetaboLights
Project description:Chronic Lymphocytic Leukemia (CLL) cells multiply in secondary lymphoid tissue but the mechanisms leading to their proliferation are still uncertain. In addition to BCR-triggered signals, other microenvironmental factors might well be involved. In proliferation centres, leukemic B cells are in close contact with CD4+CD40L+ T cells. Therefore, we here dissected the signals provided by autologous activated T cells (Tact) to CLL cells. Although the gene expression profile induced by Tact was highly similar to that induced by sole CD40 signaling, an obvious difference was that Tact induced proliferation of CLL cells. We determined that stimulation with only CD40L+IL-21 was sufficient to induce robust proliferation in CLL cells. We then defined an IL-21-induced gene signature in CLL, containing components of JAK-STAT and apoptosis pathways, and this signature could be detected in lymph node (LN) samples from patients. Finally, we could detect IL-21 RNA and protein in LN, and IL-21 production ex vivo by LN CD4+CXCR5+ follicular helper T cells. These results indicate that, in addition to BCR signaling, activated T cells might contribute to CLL cell proliferation via CD40 and IL-21. Targeting these signaling pathways might offer new venues for treatment of CLL.
2013-10-07 | GSE50572 | GEO
Project description:Microarray analysis using the HumanHT-12 v4 Expression BeadChip was utilized to determine the biological function of IL-22. The data indicate an extensive effect of IL-22 on many major molecular functions including activation of antimicrobial genes and downregulation of immune-associated pathways. Functional studies performed in-vitro using human salivary gland cells treated with IL-22 indicated a direct effect of IL-22 on cell cycling, specifically reducing cellular proliferation at the G2-M phase by activation of STAT3. Total RNAs were obtained from salivary gland cells stimulated with recombinant IL-22 for 45 minutes. Differential expression analyses were conducted using the LIMMA package from the Bioconductor project. MTT assay, flow cytometry and western blotting were used to identify the function of IL-22 on human salivary gland cells.
2015-10-27 | E-GEOD-74389 | biostudies-arrayexpress
Project description:Pulmonary arterial hypertension (PAH) is characterized by severe obstruction of small pulmonary arteries and concomitant high pulmonary arterial pressure, resulting in progressive right ventricular failure. Previously, we demonstrated that long-term interleukin (IL)-33 administration in mice induced severe occlusive arterial hypertrophy in the lung, which was mediated by group 2 innate lymphoid cells (ILC2s). In response to IL-33, ILC2s accumulated around blood vessels and produced IL-5, leading to perivascular eosinophil recruitment. In this study, we further characterized IL-33-induced pulmonary arterial hypertrophy. We first demonstrated that long-term IL-33 administration caused an increase in the right ventricular pressure. In IL-33 deficient mice, pulmonary arterial hypertrophy mediated by eggs of Schistosoma mansoni (S. mansoni) was attenuated, accompanied with partial reduction in ILC2s, eosinophils and CD4+ T cells. In addition, proteomic analysis revealed dramatic changes in urine samples from mice treated with IL-33 or S. mansoni eggs. Resistin like alpha (RELM), a pulmonary hypertension-related molecule, in the urine was commonly detected in both treatments. Large amounts of RELM were observed in the lung from IL-33-treated mice. These observations support that IL-33-induced pulmonary arterial hypertrophy is a useful model to study the mechanism underlying development of PAH and expolar biomarkers to indicate the onset of PAH.
2024-09-27 | PXD055773 | JPOST Repository
Project description:Th17 cells are enriched by sorting FR4-CD4+ T cells from SKG mice. A large number of Th17 cells also develop spontaneously when CD4+ T cells from IFN-g-deficient (IFN-g-/-) BALB/c mice are transferred to T cell-deficient RAG2-deficient (RAG2-/-) mice and subjected to homeostatic proliferation, whereas they fail to develop in similar transfer of IL-6-deficient (IL-6-/-) CD4+ T cells to IL-6-/- RAG2-/- mice. To explore the functional molecules specifically expressed by Th17 cells, we conducted Gene Microarray analysis between 10-month-old SKG FR4-CD4+ cells and age-matched BALB/c FR4-CD4+ cells, and between IFN-g-/- CD4+ cells transferred to RAG2-/- mice and IL-6-/- CD4+ T cells transferred to IL-6-/- RAG2-/- mice. The analysis revealed that 1,556 and 115 genes were up-regulated in 10-month-old SKG FR4-CD4+ and IFN-g-/- CD4+ T cells after homeostatic proliferation, respectively, with 29 genes shared by the two groups of genes. The 29 genes included those encoding cytokines, chemokines, and their receptors, such as IL-1 receptor type1 (IL-1R1), IL-17, IL-22, IL-21, CCR6, and CCL20. Keywords: cell type comparison
2007-10-17 | GSE9316 | GEO