organism: Homo sapiens
description: Protocols: Freshly isolated nasal cells and PBMCs were rested in culture medium (AIM-V supplemented with 2% AB serum) for 2 hours at 37 °C. Following the resting period, half of the cells were stimulated with anti-CD3/CD28 Dynabeads at a 1:1 cell-to-bead ratio, while the remaining half were left unstimulated as controls. Cells were cultured overnight in 96-well U-bottom tissue culture plates under standard incubation conditions (37 °C, 5% CO2). After 24 hours, both nasal and blood T cells were washed with PBS and subsequently stained with surface markers (NIR, CD3, CD4, CD8, CD45RO) and sorted using FACS (BD FACSAria III). Specifically, NIR-CD3+CD4-CD8+CD45RO+ cells were sorted from each sample for the single-cell RNA sequencing experiment. The sorted cells were labeled with distinct Hash Tag Oligonucleotides (HTOs) for multiplexing.  Single-cell suspensions were loaded onto the 10x Genomics Chromium Controller for partitioning into Gel Bead-In-Emulsion (GEMs) using the Chromium Next GEM Single Cell 5' Kit v2 (10x Genomics). Reverse transcription was performed within the GEMs, followed by amplification and library construction. HTO libraries were constructed following the TotalSeq™ protocol.  Reverse transcription was performed within the GEMs, followed by amplification and library construction. HTO libraries were constructed following the TotalSeq™ protocol.
attributes.tag: ENA first public
attributes.value: 2025-08-24
attributes.tag: INSDC center name
attributes.value: ArrayExpress
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-15436_3:Donor 2 - CSP
attributes.tag: age
attributes.value: 40
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell type
attributes.value: CD8-positive, alpha-beta T cell
attributes.tag: collection date
attributes.value: not collected
attributes.tag: common name
attributes.value: human
attributes.tag: developmental stage
attributes.value: adult
attributes.tag: disease
attributes.value: normal
attributes.tag: geographic location (country and/or sea)
attributes.value: not collected
attributes.tag: individual
attributes.value: Donor 2
attributes.tag: isolate
attributes.value: not applicable
attributes.tag: organism part
attributes.value: nasal cavity and pbmc
attributes.tag: sample name
attributes.value: E-MTAB-15436_3:Donor 2 - CSP
attributes.tag: scientific_name
attributes.value: Homo sapiens
attributes.tag: sex
attributes.value: male
attributes.tag: stimulus
attributes.value: negative or cd3/28 beads