characteristics: ERC000011
characteristics: ENA-CHECKLIST
characteristics: 2022-10-03T00:21:12Z
characteristics: ENA-FIRST-PUBLIC
characteristics: 2022-10-03T00:21:12Z
characteristics: ENA-LAST-UPDATE
characteristics: SAMEA111386510
characteristics: External Id
characteristics: Department of Dermatology, Inselspital, Bern University Hospital, University of Bern, Bern, Switzerland
characteristics: INSDC center name
characteristics: 2022-10-03T00:21:12Z
characteristics: INSDC first public
characteristics: 2022-10-03T00:21:12Z
characteristics: INSDC last update
characteristics: public
characteristics: INSDC status
characteristics: ERS13483488
characteristics: SRA accession
characteristics: E-MTAB-12204:C4R5IL9
characteristics: Submitter Id
characteristics: 42
characteristics: year
characteristics: age
characteristics: ArrayExpress
characteristics: broker name
characteristics: resting
characteristics: cell cycle stage
characteristics: CD4-positive helper T cell
characteristics: cell_type
characteristics: human
characteristics: common name
characteristics: Protocols: Isolation of T cells from blood: Peripheral blood mononuclear cells (PBMCs) were isolated by Ficoll-Plaque Plus (GE Healthcare, UK) density gradient centrifugation. Human CD4+ T cells were isolated from PBMC by EasySep positive selection kit (Stemcell Technologies) according to manufacturer’s instruction. Positively selected CD4+ T cells were washed with PBS and stained for subsequent Th cell subset sorting. Effector memory Th9 (CXCR3-CCR8+CCR6-CCR4+) and Th2 (CXCR3-CCR8-CCR6-CCR4+) were sorted to over 90% purity according to their expression of chemokine receptors from CD45RA-CD25-CD8-CD3+ cells. Single cell Th9 clones were directly sorted into 96-well plate according to their expression of chemokine receptors. After T cell single cell cloning, cells were screened for IL-9R expression by flow cytometry. Isolation of T cells from skin biopsies: Skin biopsies from positive patch test reactions were cultured in culture medium and CD4+ T cells were isolated and screened for IL-9R expression by flow cytometry. Culture medium consisted of PRMI-1640 with Hepes (GIBCO) supplemented with 5% of heat-inactivated human serum (Swiss Red Cross, Basel, Switzerland), 2mM L-Glutamine (Biochrom), 50U/ml penicillin and 50μg/ml streptomycin (Bioconcept) and 5 IU/ml IL-2 (Roche). T cells were cultured at a density of 0.25-1x105cells/96-well plate in a total volume of 200μL cell culture medium. IL-9R+ T helper cells were incubated with or without IL-9 (5ng/ml) for 12 hours. Total RNA was isolated with RNAeasy kit (Qiagen). Followed Illumina TruSeq Stranded mRNA protocol.
characteristics: description
characteristics: adult
characteristics: developmental stage
characteristics: normal
characteristics: disease
characteristics: IL-9R+
characteristics: immunophenotype
characteristics: C4
characteristics: individual
characteristics: Homo sapiens
characteristics: organism
characteristics: Homo sapiens
characteristics: organism
characteristics: blood
characteristics: organism part
characteristics: Homo sapiens
characteristics: scientific_name
characteristics: male
characteristics: sex
characteristics: C4R5IL9
characteristics: title