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Supplementary Figure 1 Correlation between TAA-specific CD8+ T-cell response breadth and patient survival. PFS data are shown for a total of 76 HCC patients after stratification based on the number of TAA targeted by CD8+ T-cells. The correlation between TAA and PFS was analyzed by logrank-test for trend.

Supplementary Figure 2 Fine-mapping of novel CD8+ T-cell epitopes. Antigen-unspecifically expanded CD8+ T-cells were either left unstimulated, stimulated with overlapping peptides or stimulated with exact epitopes. Production of IFN-γ is shown in representative dot plots. Each row represents one patient. Peptide sequences are indicated. The exact epitope in the overlapping peptide is underlined. Numbers indicate %IFN-γ+/CD8+ T-cells. (A) Epitope GPC-3519-528/HLA-A*03:01 within peptide GPC-3 65. (B) Epitope GPC-3281-289/HLA-A*02:01 within peptide GPC-3 36. (C) Epitope NY-ESO-1145-153/HLA-A*02:01 within peptide NY-ESO-1 18.

Supplementary Figure 3 Expression of cytotoxic mediators after antigen-specific expansion. After two weeks of antigen-specific expansion, TAA-specific CD8+ T-cell lines were analyzed for expression of cytotoxic mediators. Representative dot plots and histogram showing the tetramer-binding population and its expression of Granzyme B, Perforin, FasL and CD25 (black lines) compared to total CD8+ T-cells (grey) for one patient with a response to MAGE-A196-104 (A) and one patient with a response to NY-ESO-1157-165 (B). Numbers indicate %tetramer+/CD8+ T-cells. The mean fluorescence intensities (MFI) of tetramer+CD8+ T-cells are shown for Granzyme B (C), Perforin (D), FasL (E) and CD25 (F). Each dot represents one of seven TAA-specific CD8+ T-cell lines. Values were compared by paired t-test (C-E) or by Wilcoxon matched-pair test (F).

Supplementary Figure 4 Expression of inhibitory receptors after antigen-specific expansion. After two weeks of antigen-specific expansion, the expression of the inhibitory receptors programmed death-1 (PD-1) and T-cell immunoglobulin and mucin-3 (Tim-3) was analyzed on tetramer-binding CD8+ T-cells. Representative dot plots and histograms showing the tetramer-binding population and its expression of PD-1 and Tim-3 (black lines) compared to total CD8+ T-cells (grey) of one cell line specific for HLA-A*02/NY-ESO-1157-165 (A) and one specific for HLA-A*02/EBV BMLF-1280-288 (B) are shown. The percentages of PD-1+ (C) and Tim-3+ (D) among tetramer+CD8+ T-cells are shown for virus-specific and TAA-specific CD8+ T-cell lines. Comparisons were performed by Mann-Whitney U-test.

Supplementary Figure 5 Effect of inhibitory receptor blockade and cytokine supplementation on antigen-specific expansion. Antigen-specific expansion was essentially performed as described. At initiation of cultures either anti-PD-L1 mAb (A) or a mixture of IL-7 and IL-12 (B) was added. Black squares represent frequencies of tetramer-binding cells, white circles represent frequencies of IFN-γ producing cells. Each dot is representative of one cell line.

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