American Association for Cancer Research
crc-22-0514_fig2.png (149.76 kB)

FIGURE 2 from Immunologic Characterization and T cell Receptor Repertoires of Expanded Tumor-infiltrating Lymphocytes in Patients with Renal Cell Carcinoma

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posted on 2023-07-21, 17:40 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

REP TIL T cells co-cultured with tumor cells show differences in IFNγ and TNFα expressions. A, REP TILs were cocultured with the corresponding tumor cells (n = 10) either for 6 or 48 hours. The cytokine secretion of CD4+ and CD8+ T cells was analyzed with intracellular flow cytometry staining. The expression of IFNγ and TNFα moderately increased in REP TILs after the co-culture with tumor cells without additional T cell stimulation. After 48 hours co-culture, IFNγ and TNFα levels were increased in both CD4+ (0.74% vs. 0.19%, P = 0.044) and CD8+ T cells (0.78% vs. 0.13%, P = 0.025) compared with baseline REP TILs without tumor cell co-culture. No differences were observed at 6 hours co-cultures (Supplementary Fig. S3A). BASELINE_48h = only REP TILs at 48h, CO-CULTURE_48h = cocultured REP TILs without any stimulation at 48 hours. B, The immunophenotype of T cells after co-culture with tumor cells was analyzed with flow cytometry. The expression of LAG-3 moderately decreased in the CD4+ (median 13.55% vs. 1.84%, P = 0.048) and CD8+ T cells (45.55% vs. 22.94%, P = 0.029) compared with REP TIL baseline cells at 6 hours. The same trends were not observed when REP TILs were co-cultured for 48 hours (Supplementary Fig. S3D). Although not significant, the expression of PD-1 expression increased in half of the patients. BASELINE_6h = only REP TILs at 6h, CO-CULTURE_6h co-cultured REP TILs without any stimulation at 6 hours. C, T cell activation potential was assessed by stimulating the cells with anti-CD3 (OKT3), anti-CD28, anti-CD49d antibodies, and IFNγ and TNFα cytokine secretion was measured as described above. When REP TILs were stimulated and co-cultured for 6 hours, an increase in the CD3+ T cell IFNγ and TNFα expressions was observed between unstimulated and stimulated co-cultures (median 0.72% vs. 1.33%, P = 0.098). A moderate increase in CD4+ T cell IFNγ and TNFα expressions was also observed (0.46% 1.089%, P = 0.012), but not in the CD8+ T cells. CO-CULTURE_6h = cocultured REP TILs without any stimulation at 6h, TSTIM_6h = cocultured REP TILs that were T cell stimulated at 6 hours. D, Prolonged co-culture conditions (48 hours) did not lead to increased IFNγ and TNFα production in the CD3+, CD4+, and CD8+ T cell compartments. CO-CULTURE_48h = cocultured REP TILs without any stimulation at 48h, TSTIM_48h = cocultured REP TILs that were T cell stimulated at 48 hours. Non-parametric Wilcoxon matched-pairs signed rank test was used for all co-culture analyses. ns, not significant; *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.


Punainen Risti Veripalvelu (Finnish Red Cross Blood Service)

Academy of Finland (AKA)

Syöpäsäätiö (Cancerstiftelsen)

Sigrid Juséliuksen Säätiö (Sigrid Jusélius Stiftelse)

Signe ja Ane Gyllenbergin Säätiö (Signe and Ane Gyllenberg Foundation)

Relander Foundation (The Relander Foundation)

State funding university-level health research in Finland



TILs are a heterogenous group of immune cells that recognize and attack the tumor, thus are utilized in various clinical trials. In our study, we explored the TILs in patients with kidney cancer by expanding the TILs using a clinical-grade protocol, as well as observed their characteristics and ability to recognize the tumor using in-depth experimental and computational tools.