Fig. 4.

Adjuvant/tumor antigen-containing ganglioside-liposomes activate CD169-expressing moDCs and are cross-presented to antigen-specific T cells. (A) Ganglioside- (i.e., GM3) or Lewis Y-liposomes with MPLA and/or tumor-associated antigen for moDC activation or antigen presentation assay. GlcNAc, N-acetylglucosamine. (B) moDCs were incubated with MPLA-containing ganglioside-liposomes at 4 °C and washed, and IL-6 secretion after 24 h was measured by ELISA. Values are calculated as fold change over control liposome; data are mean ± SEM from five donors. (C and D) Ganglioside-liposomes containing short WT1 peptide and MPLA were loaded to moDCs and washed away, and WT1-specific CD8⁺ T cells were added. IFNγ secretion after 24 h was determined by ELISA. (C) Secreted IFNγ at different doses of liposomes are shown (data pooled from n = 6 donors). (D) Production of IFNγ after exposure to 1 µM ganglioside-liposomes normalized to control liposomes. Data are mean ± SEM from six donors. (E and F) GM3- or Lewis Y-liposome containing long gp100 peptide was loaded to moDCs, followed by 24 h coculture with gp100-specific T cell clone. IFNγ production by gp100-specific T cells (E) at different doses of liposomes (pooled from five donors) and (F) after treatment with 1 µM GM3- or Lewis Y-liposome normalized to control liposome. Data are mean ± SEM from five donors. Kruskal–Wallis multiple two-tailed t tests using a two-stage linear step-up procedure of Benjamini, Krieger, and Yekutieli, with Q = 0.05, was used (*adjusted P < 0.05, **adjusted P < 0.01).