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. 2018 Feb 14;9:253. doi: 10.3389/fimmu.2018.00253

Figure 2.

Figure 2

Liposomes are efficiently phagocyted by dendritic cells (DCs) and preserve a high viability. (A) Uptake of liposomes fluorescently labeled with lipid-conjugated fluorescent dye Oregon Green 488 1,2-dihexadecanoyl-sn-glycero-3-phosphoethanolamine by DCs. Upper panel: time course of the capture of fluorescently-labeled PS-liposomes by DCs obtained from control subjects (white circles, n = 5) and patients with type 1 diabetes (T1D) (black circles, n = 10) at 37°C (continuous line) and at 4°C (discontinuous line). Results are mean ± SEM. Comparisons between phagocytosis by control subjects DCs at 37 and 4°C showed significant differences [++++p < 0.0001, two-way analysis of variance (ANOVA)]; also, significant differences were found when comparing phagocytosis in patients with T1D at 37 and 4°C (***p < 0.001, ****p < 0.0001, Two-way ANOVA). No differences were found when comparing PS-liposomes uptake by DCs from control subjects and patients with T1D (Two-way ANOVA). Lower panel: time course of the capture of fluorescently-labeled PC-liposomes by DCs obtained from control subjects (white squares, n = 6) and patients with T1D (black squares, n = 9) at 37°C (continuous line) and at 4°C (discontinuous line). Results are mean ± SEM. Comparisons between phagocytosis by control subjects DCs at 37 and 4°C showed significant differences (++++p < 0.0001, Two-way ANOVA); also, significant differences were found when comparing phagocytosis in patients with T1D at 37 and 4°C (**p < 0.01, ****p < 0.0001, Two-way ANOVA). No differences were found when comparing PC-liposomes uptake by DCs from control subjects and patients with T1D (Two-way ANOVA). (B) Viability of DCs from control subjects (upper panel, white symbols, n ≥ 6) and patients with T1D (lower panel, black symbols, n ≥ 6) assessed by annexin V and 7aad staining. Triangles represent immature DCs (iDCs), circles represent iDCs cultured with PSA-liposomes and PSB-liposomes (PSAB-DCs), squares represent mature PSAB-DCs (mPSAB-DCs) and upside-down triangles represent mature DCs (mDCs). MDCs were induced by culture with cytokine cocktail.