biostudies-literatureVoogd LHorizon 2020 Marie Skłodowska-Curie ActionsNIAID NIH HHSDutch Research Council (NWO)National Institute of HealthBill and Melinda Gates Foundation109233https://www.ebi.ac.uk/biostudies/studies/S-EPMC10909886biostudies-literatureUnknown27(3)HLA-E molecules can present self- and pathogen-derived peptides to both natural killer (NK) cells and T cells. T cells that recognize HLA-E peptides via their T cell receptor (TCR) are termed donor-unrestricted T cells due to restricted allelic variation of HLA-E. The composition and repertoire of HLA-E TCRs is not known so far. We performed TCR sequencing on CD8⁺ T cells from 21 individuals recognizing HLA-E tetramers (TMs) folded with two <i>Mtb</i>-HLA-E-restricted peptides. We sorted HLA-E <i>Mtb</i> TM⁺ and TM^- CD8⁺ T cells directly <i>ex vivo</i> and performed bulk RNA-sequencing and single-cell TCR sequencing. The identified TCR repertoire was diverse and showed no conservation between and within individuals. TCRs selected from our single-iScience<i>Mtb</i> HLA-E-tetramer-sorted CD8⁺ T cells have a diverse TCR repertoire.PMC10909886R01 AI141315U19 AI057229R21 AI12713313259Ottenhoff THMFranken KLMCvan Meijgaarden KEDrittij AMHFHagedoorn RSRuibal PHeemskerk MHMVoogd LJoosten SALeitner JAUnen VVSteinberger PDavis MMScriba TJDingenouts CKEfalse<i>Mtb</i> HLA-E-tetramer-sorted CD8⁺ T cells have a diverse TCR repertoire.HLA-E molecules can present self- and pathogen-derived peptides to both natural killer (NK) cells and T cells. T cells that recognize HLA-E peptides via their T cell receptor (TCR) are termed donor-unrestricted T cells due to restricted allelic variation of HLA-E. The composition and repertoire of HLA-E TCRs is not known so far. We performed TCR sequencing on CD8⁺ T cells from 21 individuals recognizing HLA-E tetramers (TMs) folded with two <i>Mtb</i>-HLA-E-restricted peptides. We sorted HLA-E <i>Mtb</i> TM⁺ and TM^- CD8⁺ T cells directly <i>ex vivo</i> and performed bulk RNA-sequencing and single-cell TCR sequencing. The identified TCR repertoire was diverse and showed no conservation between and within individuals. TCRs selected from our single-2024-01-01T00:00:00Z2024 Mar2026-07-15T00:18:15.071Z2026-06-28T03:06:20.409ZS-EPMC109098863843995810.1016/j.isci.2024.109233