Figure 3.

The DC line JawsII efficiently presented multiple sulfatide isoforms to XV19 T hybridoma cells. JawsII cells (5 × 10³ cells per well) were incubated with the indicated sulfatides for 4h, followed by addition of XV19 NKT-cell hybridoma cells (40 × 10³ cells per well). After 16 h, IL-2 secretion was assayed using CTLL-2 cells. Data shown are representative of three or more experiments (mean ± SD of triplicate cultures). (A) JawsII cells had been pre-pulsed with lyso or C24:1 at the indicated concentrations, before co-culture with XV19 NKT-cell hybridoma cells. Inset: CD1d expression of JawsII cells was determined by flow cytometry (dotted line; cells stained with isotype control antibody, and solid line; CD1d–stained cells). (B) JawsII cells were incubated together with titrations of indicated sulfatide isoforms, followed by the addition of XV19 NKT-cell hybridoma cells. (C) JawsII cells (5 × 10³) were cultured together with selected sulfatide isoforms or native sulfatide (30 nmol/mL) before stimulation of the XV19 NKT-cell hybridoma. (D) Bone marrow-derived DC (10 × 10³ cells/well) were incubated with the indicated sulfatide preparations (30 nmol/mL) before stimulation of XV19 cells. Lyso, lysosulfatide.