{"database":"biostudies-arrayexpress","file_versions":[],"scores":null,"additional":{"omics_type":["Unknown","Transcriptomics","Genomics","Proteomics"],"submitter":["SOURA CHAKRABORTY"],"instrument_platform":["Illumina NovaSeq 6000","Novogene in-house automated library preparation system"],"study_type":["RNA-seq of coding RNA"],"organism":["Homo sapiens"],"species":["Homo sapiens"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/E-MTAB-16635"],"description":["Peripheral blood mononuclear cells (PBMCs) were isolated by density-gradient centrifugation using Ficoll-Paque Plus. CD8⁺ T cells were purified from PBMCs by immunomagnetic negative selection and cultured for a total of 8 days. Cells were initially activated for 72 h with plate-bound anti-CD3 (1 µg/mL) and anti-CD28 (1 µg/mL). Cells were then rested/expanded for a further 72 h in fresh ImmunoCult medium supplemented with human IL-2 (100 U/mL). Finally, cells were re-stimulated with plate-bound anti-CD3 (1 µg/mL) and anti-CD28 (1 µg/mL) in the presence of cortisol (hydrocortisone; 100 nM) or vehicle control (methanol) for 48 h. Total RNA was isolated at the end of treatment for transcriptomic profiling to quantify the glucocorticoid-driven gene expression programme in activated human CD8⁺ T cells."],"repository":["biostudies-arrayexpress"],"sample_protocol":["Nucleic Acid Extraction - Total RNA was extracted from frozen RNAlater-treated CD8⁺ T-cell pellets. Samples were homogenised using QIAshredder (Qiagen) according to the manufacturer’s instructions. RNA was purified using the RNeasy Plus Mini Kit (Qiagen), including genomic DNA removal via gDNA Eliminator columns. RNA was eluted in RNase-free water and used for downstream library preparation.","Sequencing - RNA-seq libraries were sequenced by Novogene using an Illumina platform, generating paired-end reads. Standard Illumina sequencing chemistry and quality control procedures were applied. Raw FASTQ files were delivered after base calling, filtering, and adapter trimming performed by Novogene.","Library Construction - RNA-seq libraries were generated by Novogene using their standard mRNA-seq workflow. Briefly, poly-A–enriched mRNA was isolated, fragmented, and reverse-transcribed to cDNA, followed by end repair, A-tailing, adaptor ligation, and PCR amplification. Libraries were purified, quantified, and assessed for size distribution prior to sequencing.","Sample Collection - Peripheral blood was obtained from healthy adult donors following informed consent and institutional ethical approval. Whole blood was processed immediately for PBMC isolation by density-gradient centrifugation using Ficoll-Paque Plus. CD8⁺ T cells were purified from PBMCs by immunomagnetic negative selection (STEMCELL Technologies). Purified CD8⁺ T cells were cultured in ImmunoCult-XF T Cell Expansion Medium supplemented with gentamicin (50 µg/mL) for a total of 8 days. Cells were activated for 72 h with plate-bound anti-CD3 (BioLegend, OKT3, 1 µg/mL) and anti-CD28 (BioLegend, CD28.2, 1 µg/mL), rested/expanded for 72 h in fresh medium containing human IL-2 (PeproTech, 100 U/mL), then re-stimulated for 48 h with plate-bound anti-CD3/anti-CD28 in the presence of hydrocortisone (Sigma; 100 nM) or vehicle control (methanol; Sigma). At the end of treatment, cell pellets were overlaid with 40 µL RNAlater (Invitrogen) and stored at −80°C prior to RNA extraction."],"figure_sub":["Organization","MINSEQE Score","Assays and Data","MAGE-TAB Files"],"pubmed_authors":["SOURA CHAKRABORTY"],"additional_accession":[]},"is_claimable":false,"name":"Cortisol (Hydrocortisone) Treatment of Activated Primary Human CD8⁺ T Cells","description":"Peripheral blood mononuclear cells (PBMCs) were isolated by density-gradient centrifugation using Ficoll-Paque Plus. CD8⁺ T cells were purified from PBMCs by immunomagnetic negative selection and cultured for a total of 8 days. Cells were initially activated for 72 h with plate-bound anti-CD3 (1 µg/mL) and anti-CD28 (1 µg/mL). Cells were then rested/expanded for a further 72 h in fresh ImmunoCult medium supplemented with human IL-2 (100 U/mL). Finally, cells were re-stimulated with plate-bound anti-CD3 (1 µg/mL) and anti-CD28 (1 µg/mL) in the presence of cortisol (hydrocortisone; 100 nM) or vehicle control (methanol) for 48 h. Total RNA was isolated at the end of treatment for transcriptomic profiling to quantify the glucocorticoid-driven gene expression programme in activated human CD8⁺ T cells.","dates":{"release":"2026-02-18T00:00:00Z","modification":"2026-02-18T08:46:31.644Z","creation":"2026-02-11T11:20:43.478Z"},"accession":"E-MTAB-16635","cross_references":{"ENA":["ERP188989"],"EFO":["EFO_0002944","EFO_0004170","EFO_0005518","EFO_0003738","EFO_0004184"]}}