Sulfatide reduces the capacity of DCs to induce proliferation of GAD65-specific Th1 cells and diverts LPS-induced inhibition of IL-10 production by the GAD65-specific clone.

<p>(A) GAD65-specific cells were incubated for 4 days with GAD65 protein-pulsed immature or LPS or LPS+IFNγ-matured DCs which were not exposed to self-glycolipids (white bars), modulated with β-GalCer (grey bars) or with sulfatide (black bars). 3H thymidine was added during the last 18hrs of the culture. Graphs show proliferation of GAD65-specific cells (polyclonal response – Th1 T cell line; monoclonal response – PM1#11 clone), measured by 3H thymidine incorporation. The results are presented as mean counts per minute (CPM) ± SD of triplicate cultures. (B) The cytokine production by the GAD65-specific T cell clone (PM1#11) as measured by ELISpot analysis. T cells were incubated with DCs (immature, LPS-matured or LPS+IFNγ-matured) pulsed with GAD65 protein, which were untreated (–-) or modulated with β-GalCer or Sulfatide. Dark grey bars depict IL-10, light hatched bars depict IFNγ and square-hatched bars depict IL-13 production. The data show mean number of spots/10⁴ T cells ± SD of triplicate cultures. Three independent experiments were performed (n = 3).