Fig. 7: Tumor deglycosylation affects DC metabolism and function.

Sialylated and fucosylated motifs (“w/o sTn-Fuc”) or all N-linked and O-linked oligosaccharides motifs (“all deglyco”) were removed from tumor cells using either the α(2 → 3,6,8,9) Neuraminidase (sialidase) and the α-1 → (2,3,4)-Fucosidase or mixture of the endo-α-N-Acetylgalactosaminidase, β1,4-Galactosidase, Glucosaminidase, N-Glycosidase F and Neuraminidase. PanDCs (mixture of cDC2s, cDC1s and pDCs) purified from HD blood were then co-cultured or not with deglycosylated or not tumor cells, and further stimulated or not with a mixture of TLR-L. Their metabolism was then studied by flow cytometry using the SCENITH method, and their cytokine/chemokine production was analyzed in the supernatants using Luminex. A Glycolytic or mitochondrial metabolic dependencies/capacities of cDC2s and pDCs upon culture (colored symbols) or not (white symbols) with deglycosylated (or not [pink symbols]) tumor cells in unstimulated conditions (n = 4 donor/tumor line couples for sTn/Fuc removal (purple symbols); n = 6 donor/tumor line couples for total deglycosylation (blue symbols)). The central line indicates median, the box contains the 25th to 75th percentile, and whiskers the min and max values. P-values were calculated using non-parametric paired two-way ANOVA (or a mixed effect model) followed by Sidak’s multiple comparison post-tests. Cytokine/chemokine production upon co-culture or not (white symbols) of DCs with deglycosylated (dark pink) or not (light pink) tumor cells lacking sialylated and fucosylated motifs (B, n = 4 donor/tumor line couples) or all N-linked and O-linked oligosaccharides motifs (C, n = 6 donor/tumor line couples). P-values were calculated using one-way Friedman test with Dunn’s multiple comparison test. Only significant statistics are displayed on graphs. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001. Source data are provided as a Source Data file.