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Fig. 2 | Journal for ImmunoTherapy of Cancer

Fig. 2

From: A new synthetic toll-like receptor 1/2 ligand is an efficient adjuvant for peptide vaccination in a human volunteer

Fig. 2

Impact of XS15 on cytokine release by slanMo and their capacity to stimulate WT1 peptide-specific CD8+ T cells and NK cells. (a) slanMo were maintained for 6 h to allow spontaneous maturation into DCs. Subsequently, slanMo were cultivated alone (slanMo) or in the presence of XS15 (slanMo + XS15) for additional 18 h. Supernatants were collected and the concentration of (a) TNF (left), IL-1β (middle), IL-6 (right), IL-23 (lower left) analysed by ELISA. (b) slanMo were maintained for 6 h to allow spontaneous maturation into DCs. Subsequently, slanMo were cultivated in the absence (slanMo) or presence of XS15 (slanMo + XS15) for additional 18 h, alternatively, slanMo were incubated with IFNγ for the first 6 h. Thereafter, slanMo were cultivated in the presence of IFNγ alone (slanMo + IFNγ) or together with XS15 (slanMo + IFNγ + XS15) for additional 18 h. Then, IL-12 was analysed by ELISA. The results of three different healthy donors (HD) are presented as mean ± SE of duplicate or triplicate measurements. (c) Effect of XS15 on the capacity of slanMo to stimulate IFNγ release by WT1 peptide-specific CD8+ T cells: slanMo were maintained for 6 h to allow spontaneous maturation. Subsequently, slanMo were coincubated with the specific CD8+ T cell clone CC7 (slanMo + CD8+), in the presence of the WT1 peptide (WT1) and/or XS15. After 42 h, supernatants were collected and IFNγ was quantified by ELISA. The results of three different healthy donors (HD) are presented as mean ± SE of triplicate determinations. (d) Impact of XS15 on the ability of slanMo to stimulate IFNγ secretion by NK cells: slanMo were maintained for 6 h to allow spontaneous maturation. Then, autologous NK cells were cultured either alone (NK) or incubated with XS15 (NK + XS15), cocultured with slanMo alone (NK + slanMo) or additionally incubated with XS15 (NK + slanMo +XS15). After 42 h, supernatants were collected and the concentration of IFNγ was determined by ELISA. The results of three different HD are presented as mean ± SE of triplicate determinations. Asterisks indicate a statistically significant difference (*p ≤ 0.05; **p ≤ 0.01; ***p ≤ 0.001; assessed by Student’s t-test). Exemplary flow cytometry results showing effects of XS15 on slanMo-mediated T cell programming regarding the percentage of IFNγ- and IL-4-producing CD4+ T cells are provided as Additional file 8: Fig. S1

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