biostudies-arrayexpressCarolin TurnerHomo sapienshttps://www.ebi.ac.uk/biostudies/studies/E-MTAB-14246We undertook single cell RNA, TCR and antibody-derived tag (ADT) sequencing of human skin suction blisters induced at the site of of the tuberculin skin test on day 2. This submission comprises data from 31 individuals (9 male, 22 female) with immunological memory to mycobacterial antigens. TotalSeq-C Human Universal Cocktail v1 (BioLegend) was used to tag 130 cell surface proteins. Data were acquired with 10X Genomics technology. Following quality control filtering, the integrated dataset contained 63881 cells, with concurrent TCR sequencing data for 37413 cells.biostudies-arrayexpressSequencing - Sequencing was performed on Illumina’s NovaSeq6000 system, using paired end 150 bp reads, and targeting 20,000 read pairs per cell for gene expression libraries, and 5,000 read pairs per cell for TCR VDJ and feature barcode libraries respectively.Nucleic Acid Extraction - Cells were stained with the TotalSeq-C Human Universal Cocktail v1 (BioLegend). Nucleic acids were extracted following the manufacturer’s instructions for the Chromium SingleCell 5’ Reagent kit v2 (10x Genomics).Library Construction - Library construction was performed using the Chromium SingleCell 5’ Reagent kit v2 (10x Genomics) according to the manufacturer’s instructions to generate gene expression, T cell receptor (TCR) VDJ, and surface protein (= feature barcode) libraries.Sample Collection - Participants received intradermal injection of 0.1 ml 2U tuberculin in the volar aspect of the forearm. At 48 hours, suction blisters were induced at the injection site through the application of negative pressure and blister fluid aspirated 2-4 hours later. Blister cells were recovered after centrifugation of blister fluid.OrganizationMINSEQE ScoreAssays and DataProcessed DataMAGE-TAB FilesData Transformation - CellRanger's filtered output data are provided as processed data files. Nor further data processing or normalisation was performed.Sequence Alignment - Read alignment, feature counting and cell calling was performed with 10x Genomics CellRanger (v7.1.0) against the human genome assembly GRCh38 (gene expression reference version 2020-A and VDJ-T reference version 7.1), using the ‘multi’ pipeline.MetabolomicsUnknownTranscriptomicsGenomicsProteomicsIllumina NovaSeq 6000The tuberculin skin test (TST) is a cutaneous delayed hypersensitivity reaction to antigen from Mycobacterium tuberculosis (Mtb). We provide the first single cell sequencing characterisation of the human TST reaction, based on skin suction blisters induced at the site of the TST on day 2 in 31 individuals. Integrated single cell RNA and TCR sequencing showed the immune response to be dominated by T cells, with smaller populations of NK cells and myeloid cells. T cells comprised CD4, CD8, gamma/delta and NK T cells, with 50% of all T cells identified as cytotoxic and 14% as regulatory. Interferon gamma gene expression was strongest in CD8 T cells, and distinct CD4 T helper lineages could not unambiguously be identified at this time point. Amongst myeloid cells, 63% displayed antimicrobial RNA-seq of coding RNA from single cellsHomo sapiensSingle-cell transcriptome and T cell receptor profiling of the tuberculin skin testCarolin T Turner, Joshua Rosenheim, Clare Thakker, Aneesh Chandran, Holly Wilson, Cristina Venturini, Gabriele Pollara, Benny Chain, Gillian S Tomlinson, Mahdad NoursadeghiCarolin TurnerfalseSingle-cell transcriptome and T cell receptor profiling of the tuberculin skin testWe undertook single cell RNA, TCR and antibody-derived tag (ADT) sequencing of human skin suction blisters induced at the site of of the tuberculin skin test on day 2. This submission comprises data from 31 individuals (9 male, 22 female) with immunological memory to mycobacterial antigens. TotalSeq-C Human Universal Cocktail v1 (BioLegend) was used to tag 130 cell surface proteins. Data were acquired with 10X Genomics technology. Following quality control filtering, the integrated dataset contained 63881 cells, with concurrent TCR sequencing data for 37413 cells.2026-07-09T00:00:00Z2026-07-09T07:57:03.713Z2024-07-19T14:32:29.285ZE-MTAB-14246ERP162148EFO_0002944EFO_0004170EFO_0005684EFO_0004917EFO_0005518EFO_0003816EFO_000418410.1101/2024.06.25.600676