Figure 7.

Bacterial glycolipids stimulate macrophage-dependent iNKT cell activation (a-b) Fold increase in total cell number (a) or in the depicted populations (b) in LNs of WT (a,b) and J_α18 (NKT KO, a) mice after injection with GSL-1′ particles relative to animals injected with control particles, *p<0.05; NS, no significant (c-f) iNKT cell activation was assessed by flow cytometry after i.p. injection of GSL-1′ (black) or control particles (grey). (c,d) CD25 expression in mediastinal LN iNKT cells at various times after injection. MFI were normalized to mice injected with control particles. (e,f) Expression of CD69 (e) and intracellular IFN-γ (f) in iNKT cells from mediastinal LNs 12 h after injection. Data are representative of 3 independent experiments. (g) CD25 expression in LN iNKT cells was assessed 12 h after injection of GSL-1′ particles into CLL treated (red) or untreated (black) mice. MFI were normalized to mice injected with particles containing control lipids. Black lines indicate mean values. Data points represent one mediastinal LN from individual mice and representative from two independent experiments with at least 2 animals per experiment. Data were compared with two-tailed t-test, ***p=0.0006. (h-k) iNKT-S1 (red) and CD4⁺T cells (blue) were transferred into WT recipients injected i.p. with GSL-1′ particles. Average speed (h), instantaneous speed distribution (i), arrest coefficient (j) and mean displacement (± s.e.m., k) for iNKT and CD4⁺T cells are shown at 16 h after antigen administration. Data were obtained from at least two independent experiments and compared with a two-tailed unpaired Mann-Whitney test. **p<0.005

Error bars (a,b), s.e.m.