American Association for Cancer Research
crc-22-0514-s10.pptx (416.62 kB)

Figure S5 from Immunological characterization and T-cell receptor repertoires of expanded tumor-infiltrating lymphocytes in renal cell carcinoma patients

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posted on 2023-06-26, 14:20 authored by Moon Hee Lee, Jason Theodoropoulos, Jani Huuhtanen, Dipabarna Bhattacharya, Petrus Järvinen, Sara Tornberg, Harry Nísen, Tuomas Mirtti, Ilona Uski, Anita Kumari, Karita Peltonen, Arianna Draghi, Marco Donia, Anna Kreutzman, Satu Mustjoki

Figure S5 shows representative gating strategies for the co-culture assays with different timepoints (6h, 48h) and conditions (baseline, unstimulated, T-cell stimulated).



The successful use of expanded tumor-infiltrating lymphocytes (TILs) in adoptive TIL therapies has been reported, but the effects of the TIL expansion, immunophenotype, function, and T-cell receptor (TCR) repertoire of the infused products relative to the tumor microenvironment (TME) are not well understood. In this study, we analyzed the tumor samples (n=58) from treatment-naïve RCC patients, “pre-rapidly expanded” TILs (pre-REP TIL, n=15) and “rapidly expanded” TILs (REP TILs, n=25) according to a clinical-grade TIL production protocol, with scRNA+TCRab-seq, TCRb-seq, and flow cytometry. REP TILs encompassed a greater abundance of CD4+ than CD8+ T-cells, with increased LAG-3 and low PD-1 expressions in both CD4+ and CD8+ T-cell compartments compared to the pre-REP TIL and tumor T-cells. The REP protocol preferentially expanded small clones of the CD4+ phenotype (CD4, IL7R, KLRB1) in the TME, indicating that the largest exhausted T-cell clones in the tumor do not expand during the expansion protocol. Additionally, by generating a catalog of RCC-associated TCR motifs from >1000 scRNA+TCRab-seq and TCRb-seq RCC, healthy and other cancer sample cohorts, we quantified the RCC-associated TCRs from the expansion protocol. Unlike the low-remaining amount of anti-viral TCRs throughout the expansion, the quantity of the RCC-associated TCRs was high in the tumors and pre-REP TILs but decreased in the REP TILs. Our results provide an in-depth understanding of the origin, phenotype, and TCR specificity of RCC TIL products, paving the way for a more rationalized production of TILs.