Divergent gene expression and kinetics of circulating tumor-infiltrating lymphocytes with highly homologous T-cell receptor sequences during immunotherapy

single-cell RNA/TCR sequencing (scRNA/TCR-seq) of circulating T cells from longitudinally obtained blood samples from the same a patient with NSCLC who had a response and a subsequent progression to anti-PD-1/CTLA-4 blockade therapy. Single live CD45+ CD3+ cells were enriched from cryopreserved PBMC samples obtained pre-treatment and on-treatment at 4, 8, 12, 24, and 32 weeks by flow cytometry sorting using a BD FACSAria II (BD Biosciences).