T-cell receptor sequences from ECCITE-seq of BNT162b2 mRNA-Vaccinated Donor PBMCs
For ECCITE-seq experiment, sorted cells were centrifuged at 400 RCF for 8 min at 4 °C and then resuspended in staining buffer. TotalSeq-C human cocktail (BioLegend) was added for the surface protein staining, on ice for 30 min. After washing three times with 1 ml staining buffer, cells were resuspended in PBS and the cell concentration was adjusted to about 2000 cells/µL. Cells were loaded onto the 10x Chromium Next GEM Chip N, following manufacturer recommendations (Chromium Next GEM Single Cell 5' HT Reagent Kits v2). During cDNA amplification, 0.2 μM each of ADT (5′CCTTGGCACCCGAGAATT∗C∗C) and HTO (5′GTGACTGGAGTTCAGACGTGTGC∗T∗C) were added to the reaction. Gene expression (RNA), hashtags (HTOs), T-cell receptor sequencing (TCR), and oligo-tagged antibodies (ADTs) libraries were constructed by following 10x Genomics and ECCITE-seq protocols.