Sample 13
Homo sapiens
Protocols: After three days in culture, CD3+ Vδ2+ T-cells were enriched by cell sorting using a FACSAria II (BD Biosciences), to purities >99.8%. Monocytes were freshly isolated and irradiated at 50 Gy prior to co-culture with freshly isolated autologous at a monocyte to γδ T-cell ratio of 1:10.  Cultures were treated with 10 nM HMB-PP alone, or in combination with 100 U/ml IL-2, 20 ng/ml IL-15 or 20 ng/ml IL-21.  RNA was extracted from cell pellets using the RNeasy Micro Kit (Qiagen) and examined qualitatively and quantitatively using the TapeStation 2200 system (Agilent) and a Qubit fluorometer (Thermo Fisher Scientific).  Ribosomal RNA was depleted as part of the Illumina Stranded Total RNA Prep protocol using coated magnetic beads; the remaining RNA was fragmented and denatured, and first and second strand cDNA synthesis and anchor ligation was carried out according to the manufacturer's instructions.
replicate
TR02
individual
D1
organism
Homo sapiens
organism
Homo sapiens
collection date
not collected
scientific_name
Homo sapiens
common name
human
organism part
blood
developmental stage
adult
isolate
not applicable
growth condition
irradiation and co-culture with gamma-delta T cells
cell_line
PBMC
cell_type
monocyte
sequencing run
AH33YJBGXY
geographic location (country and/or sea)
not collected