biostudies-arrayexpressUnknownTranscriptomicsGenomicsProteomicsGong YuIllumina NovaSeq 6000RNA-seq of coding RNA from single cellsMus musculusMus musculushttps://www.ebi.ac.uk/biostudies/studies/E-MTAB-16148This study aims to investigate the functional role of Ly49+CD8+T cells in allogeneic immune responses during cardiac transplantation. Using a murine heart transplant model and mixed lymphocyte reactions, we analyzed how these cells respond under CD80,86-CD28 co-stimulation blockade. The results revealed that Ly49+CD8+T cells exhibit distinct proliferative and cytotoxic features and contribute to immune regulation through Qa-1 interaction and inhibitory receptors such as LAG-3 and CTLA-4. This research provides new insights into the immunoregulatory mechanisms that promote graft tolerance and may inform future strategies to prevent transplant rejection.biostudies-arrayexpressNucleic Acid Extraction - After a single freeze and thawing cycle, cDNAs were synthesized from whole cell lysate and amplified by one-step RT-TS-PCR method (Repertoire Genesis, Osaka, Japan) using PrimeScriptⅡ High Fidelity One-Step RT-PCR Kit (Takara Bio, Kusatsu, Japan), adapter-added oligo dT primers, adaptor-added template-switch oligos and adaptor primers.Sample Collection - B6 splenocytes were obtained from naïve mice or after MLR in the presence or absence of anti-CD80/86 mAb. Anergic B6 splenocytes obtained after MLR with anti-CD80/86 mAb were added into MLR of Ly49+ cell-depleted B6 splenocytes with irradiated LPS-stimulated BALB/c splenocytes, then live cells were prepared on day 3 and stained with FITC-conjugated anti-mouse CD3 mAb (17A2), APC/Cy7-conjugated anti-mouse CD8α mAb (53-6.7), and PE-conjugated anti-mouse Ly49 mAb (14B11).Single Ly49+ CD8+ T cell was dropped into every well on 96 well plates preloaded with lysis buffer containing NP-40 and RNase inhibitor using FACSmelody (BD Biosciences).Sequencing - . The libraries were sequenced on Illumina NovaSeq 6000 (Illumina) using a 2 x 150 paired end configuration.Library Construction - Sequencing libraries were generated using KAPA HyperPlus Kit (Roche Diagnostics, Basel, Switzerland) and NEBNext Multiplex Oligos for Illumina (New England Biolabs) following the manufacturer’s recommendations.OrganizationMINSEQE ScoreAssays and DataMAGE-TAB FilesGong YufalseSingle-cell RNA sequencing data of Ly49⁺ CD8 T cells treated with anti-CD80/86 antibody and control IgThis study aims to investigate the functional role of Ly49+CD8+T cells in allogeneic immune responses during cardiac transplantation. Using a murine heart transplant model and mixed lymphocyte reactions, we analyzed how these cells respond under CD80,86-CD28 co-stimulation blockade. The results revealed that Ly49+CD8+T cells exhibit distinct proliferative and cytotoxic features and contribute to immune regulation through Qa-1 interaction and inhibitory receptors such as LAG-3 and CTLA-4. This research provides new insights into the immunoregulatory mechanisms that promote graft tolerance and may inform future strategies to prevent transplant rejection.2025-12-06T00:00:00Z2025-12-06T02:02:15.921Z2025-11-22T20:39:56.882ZE-MTAB-16148ERP185507EFO_0002944EFO_0004170EFO_0005684EFO_0005518EFO_0004184