Figure 2

S1P induces the migration of T and NKT cells through S1P₁, whereas FTY720 inhibits the migration of T cells but not of NKT cells. (a) Mononuclear cells obtained from the livers of B6 mice were pre-incubated with FTY720p (1 μg), or PTX (50 ng), and then washed with PBS. These cells were incubated in the upper chamber of a transwell system, whereas 600–800 μl of media containing 1 μg/ml of S1P or SEW2871 was placed in the bottom chamber. After 12 h, the cells from the bottom chamber were removed and stained with anti-NK1.1 and TCR-β mAbs. (b–d) Sorted NK1.1⁺TCR-β⁺ NKT and NK1.1⁻TCR-β⁺ T cells were pre-incubated with or without FTY720p (1 μg) for 30 min and then washed with PBS. These cells were incubated in the upper chamber of a transwell system, whereas S1P (1 μg/ml) was placed in the bottom chamber. After 24 h (panels b and c) or 12 h (panel d), the cells were taken from the upper and bottom chambers, and analyzed by flow cytometry after staining with anti-NK1.1 and TCR-β mAbs (panels b and c), or without staining with these antibodies (panel d). (Panels a, b, and d) The percentage of migration of the T and NKT cells is shown. (Panels a–d) The results shown are representative of three independent experiments. (NS, not significant; ^*P<0.05 and ^**P<0.01).