Fig. 7: Identification of paired αβ TCR sequences with their antigen specificities.
From: High throughput pMHC-I tetramer library production using chaperone-mediated peptide exchange
a Recovery of MART-1 tetramer barcodes on DMF5 cells from PBMC-DC co-cultures spiked with 1% DMF5 cells. Number of MART-1 tetramer reads among DMF5 positive (DMF5-TCR) and negative (NB-TCR) cells. Cells are classified as positive/negative according to sequencing reads of the DMF5 TCR, where >10 DMF5 reads was used as a cutoff to classify DMF5+ cells. Box graphs display the distribution of MART-1 tetramer reads from DMF5+ (>10 DMF5 reads n = 76) and DMF5- (≤10 DMF5 reads, n = 927) cells. Upper/lower bars and box boundaries indicate the 95th/5th and 75th/25th percentiles, respectively, horizontal box lines indicate medians. Statistical significance was assessed using a two-tailed Mann-Whitney test (p-value < 0.0001). b Distribution of antigen specificities identified from tetramer + /CD8+ T cells from human splenocytes and the number of tetramer-barcode read per cell. Each dot represents a single cell, n = 102 in total, error bars correspond to one standard deviation from the mean. c, d V(D)J usage of cells identified as specific for the EBV-BRLF1 (YVLDHLIVV) and NY-ESO-1 (SLLMWITQA) antigens. All TRAVJ (n = 11/35) and TRBVJ (n = 14/35) chains identified are represented. e TCR CDR3 sequences identified for antigen-specific T cells. Consensus BRLF-1 TRA and NY-ESO-1 TRB chains are highlighted in orange and red, respectively. Gating strategies used for sorting tetramer+ cells are outlined in Supplementary Fig. 11. Source data are provided as a Source Data file.
