Figure 1.

Induction of T-cell responses to SVX in healthy donors and spontaneous T-cell responses in cancer patients. (A–D) CD4⁺ and CD8⁺ T-cell responses against SVX peptides in healthy donors. CD4⁺ T cells from 12 healthy donors with diverse HLA-DRB1 genotype (A,B) or CD8⁺ T cells from 3 HLA-A*02:01 healthy donors (C,D) were repeatedly stimulated in vitro with the pool of SVX peptides (S1+S2+S3) loaded on autologous DCs. T-cell specificity was assessed by IFN-γ ELISpot assays using PBMCs or C1R-A2 loaded with the pool or individual SVX peptides. Histograms show the frequency of responding donors (A,C) and the mean percentages ± SEM of specific T-cell lines induced per donors (B,D) responding to each individual SVX peptide or at least one SVX peptide (Pool). (E,F) Spontaneous T-cell responses to SVX peptides in healthy donors, and cancer patients. (E) PBMC from 3 healthy donors, 11 head and neck, 10 lung, and 14 mRCC cancer patients were screened for spontaneous T-cell reactivity against the pool of SVX peptides, in IFN-γ ELISpot assays, after 1 week of in vitro culture with the pool of SVX peptides. (E) Intensity of the survivin response in different cancer patients and healthy donors. Each bar represents one patient or donor. Data are presented as means of IFN-γ spots from one experiment in triplicate. Small histograms represent the percentage of responder and of non-responder in healthy donors and in different types of cancer patients. (F) The 11 mRCC-responder patients were screened for spontaneous T-cell reactivity against individual SVX peptide, in IFN-γ ELISpot assays, after 1 week of in vitro culture. Data are presented as means of IFN-γ spots from one experiment in triplicate. A response was considered positive if the number of spots per well obtained in peptide(s) stimulated conditions was two-fold higher than the number of spots counted without peptide(s), with a cut-off at 10 spot-forming cells after subtracting background.