GSM6430104: I757, replicate 1, scRNAseq; Homo sapiens; RNA-Seq
Blood from unexposed donors was collected from healthy individuals registered at the Guangzhou Blood Center until 2019. The donors had no known history of any significant systemic diseases, including, but not limited to, hepatitis B or C, HIV, diabetes, kidney or liver diseases, malignant tumors, or autoimmune diseases. The samples were confirmed by negative reports of SARS-CoV-2 RNA real-time reverse transcription polymerase chain reaction (RT-PCR) assays. PBMCs were isolated from HLA-A2+ healthy donors using EasySep Human negative selection (Stemcell, Cat#17953, Canada) according to the manufacture instruction. Each type of epitope specific CD8 T cells was generated with the method described above. Each epitope-specific CD8 T cells were labeled with PE conjugated corresponding epitope based tetramers and APC conjugated anti-CD8 antibody, and sorted by flow cytometer FACS Aria (BD). Hashtags were used to label different epitope specific CD8 T cells (Table S2). Cell number and viability were checked after surface protein hashtag staining (cell viability > 80%). Then droplet-encapsulation single-cell sequencing experiments were performed, and 10,000 living single cells were loaded onto each of the Chromium Controller (10x Genomics). After droplet-encapsulation, single-cell cDNA synthesis, amplification and sequencing libraries were generated using Chromium Single Cell 5' Feature Barcode Library Kit (10x Genomics), Chromium Single Cell 5' Library & Gel Bead Kit (10x Genomics), and Chromium Single Cell V(D)J Enrichment Kit (Human T Cell, 10x Genomics) according to the manufacturer's instructions. The libraries from each loaded channel (up to eight channels) were multiplexed together and sequenced on an Illumina Novaseq 6000.