Single-cell gene expression sequencing of peptide stimulated T cell cultures derived from FLC TILs
TILs were expanded ex vivo from tumor fragments from patient SJ2 according to Dudley et al, J Immunother, 2003. After expansion, SJ2 TILs were stimulated with 24mer DNAJB1-PRKACA fusion peptide (RKREIFDRYGEE-VKEFLAKAKEDF, 1 uM). After stimulation, TILs were stained with human hashtag/barcoded antibodies to distinguish between separate TIL wells. TILs were then subjected to scRNAseq using the 10x Chromium platform using the NextGEM Single Cell V(D)J v1.1 with Feature Barcoding kit. Antibody barcode identities include HHash3_TotalC0253 with sequence TTCCGCCTCTCTTTG, HHash4_TotalC0254 with sequence AGTAAGTTCAGCGTA