Fig 2: IL-2 promotes the proliferation, cytokine production, but reduces PD-1 expression in effector CD8+ T cells and PD-L1 in PTC cells. Effector CD3+CD8+CD25+T cells were co-cultured with the indicated PTC cells that had been pre-treated with, or without, IL-2 (0.1 µg/µL) for 24 h and the frequency of CD3+CD8+CD25+T cells and the levels of PD-1 expression were determined by flow cytometry. The levels of IL-2 in the supernatants were analyzed by ELISA and the relative levels of PD-L1 expression in PTC cells were analyzed by qRT-PCR and Western blot. Furthermore, the PD-L1 expression in PTC and PTC+HT tissues and their para-non-tumor tissues was analyzed by IHC. A, The frequency of CD3+CD8+CD25+T cells. B, The levels of IL-2. C, PD-1 expression in effector CD8+T cells. D and E, PD-L1 expression in PTC cells. F, IHC analysis of PD-L1 expression in tissues. G, Immunostaining scores of PD-L1 expression. H, The correlation between the immunostaining scores of IL-2 and PD-L1. Datas are representative images (magnification x 200 with a 400 inserter) or expressed as the mean ± SD of each group. *P<0.05, **P<0.01, ***P<0.001.

Fig 3: Cis versus trans CD137-costimulation by CD137L-Fc coated beads on human CD8+ T lymphocytes.a, schematic representation of microbeads coated with CD137L-Fc used to stimulate CD8+ T cells. Human primary CD8 T cells from healthy donors were activated with microbeads for 96 h. b, representative dot plots of cell-surface CD25 (n = 11) and c, representative histograms (n = 10) of geometric mean fluorescence intensity (gMFI) for CD25 surface expression on CD8+ T cells. d, intracellular Bcl-xL (n = 8) expression analysed by flow cytometry in the indicated conditions. Summary data are given as delta (?), the difference of the value between cis and trans conditions to which the anti-CD3 background has been subtracted in each case. e, concentrations of human IFN? (n = 5) in the culture supernatants of the indicated conditions. Data are given as mean ± s.e.m. Statistical significance was determined with the Wilcoxon test (two-sided). Source data are provided as Source Data file.

Fig 4: IL-10 promotes T-cell activation in cocultures of CD3+CD8+CD25+ T cells and PTC cell lines. (A) Flow cytometry results of the CD25+ fraction of CD3+ T cells in coculture systems containing activated CD3+CD8+CD25+ T lymphocytes (effector cells, E) and pretreated PTC cells (K-1 and TPC-1 treated with 0.1 µg/µL of IL-10 for 24 hours; target cells, T) at various E:T ratios (30:1 and 10:1) (*P < 0.05, ***P < 0.001). (B) IL-2 production in coculture systems (details described above) measured by ELISA (*P < 0.05). (C) IFN-? production in coculture systems of CD3+CD8+CD25+T lymphocytes and IL-10-stimulated PTC cells (K-1 and TPC-1) at a 30:1 ratio (***P < 0.001). (D) Mean fluorescence intensity (MFI) of PD-1 expression in coculture systems of activated CD3+CD8+CD25+ T lymphocytes and IL-10 pretreated PTC cells at a 30:1 ratio (*P < 0.05). All tests were conducted in triplicate. ELISA, enzyme-linked immunosorbent assay; H, hours; IFN, interferon; IL, interleukin; PD-1, programmed death-1; PTC, papillary thyroid cancer.

Fig 5: SLFN11 is a dual biomarker capturing simultaneously interconnected immunological and cancer cell–intrinsic functional dispositions associated with sensitivity to platinum treatment.(A) Bubble chart representing significantly enriched pathways during treatment with cisplatin in SLFN11hi (n = 3) versus SLFN11lo (n = 4) ovarian cancer cell lines. The bubble size indicates the q value (FDR), whereas the color represents the direction of the change as normalized enrichment score (NES), with red indicating a positive enrichment of the comparison and blue a negative one. c, concentration. (B) Heatmap of log fold changes of selected immune-related transcripts for the comparison between the SLFN11hi and SLFN11lo cancer cell lines at the same time points as for A. (C) PBMCs were incubated with IFN-ß, GM-CSF, or anti-CD3/CD28 beads for 24 hours and immunoblotted as indicated. Chart shows SLFN11 densitometry (n = 4). (D) DU145 parental or SLFN11-KO cells were stained with/without anti-SLFN11 and analyzed by flow cytometry. (E) PBMCs were treated as in C and analyzed by flow cytometry. FACS plots show SLFN11 and CD25 expression in naive (top) and anti-CD3/CD28-stimulated (bottom) cultures. Charts show quantification of SLFN11hi CD4+ and CD8+ T cells (top) and CD25 expression in SLFN11hi/SLFN11lo T cells (bottom). (F) Purified CD3+ T cells were stimulated with anti-CD3/CD28 beads and immunoblotted as indicated. Chart shows SLFN11 densitometry (n = 4). (G) PBMCs were treated as in C and analyzed by flow cytometry for SLFN11 and programed cell death ligand 1 (PD-L1) expression in myeloid cells. Chart shows quantification of SLFN11hi myeloid cells (n = 8). The box plots depict the minimum and maximum values (whiskers), the upper and lower quartiles, and the median. The length of the box represents the interquartile range. All data in graphs are presented as mean ± SEM. Statistics by Student’s t test, pairwise comparisons with the 2-tailed t test with reported P values adjusted for family-wise error using the Holm’s method. *P < 0.05; **P < 0.01; ****P < 0.0001.