(A) Induction of CCL20 level in plasma from uGD patients and healthy controls. Medium: RPMI 1640; NP, normal plasma; NP+ OPN: normal plasma added with 0.1 µg/ml OPN; PP, patient plasma; PP+OPNmAb : patient plasma pretreated with 5 µg/ml OPN-neutralizing antibody for 30 min. *, P<0.05 versus medium control; #, P<0.05 versus Ctrl mAb. (B, C) OPN induced CCL20 mRNA expressions (B) and protein levels (C) in a time and dose-dependent manner. For time course, the PBMCs were cultured in the presence of human recombinant OPN (1 µg/ml) and analyzed at the indicated time points. To determine the dose dependent manner of CCL20 expression, the PBMCs were cultured for 12 hours with OPN at the indicated concentrations. CD4+T cells and culture medium were then purified and analyzed. Shown are representative results from 3 independent experiments with separate specimens. All data were presented as mean±SEM. *, P<0.05; **, P<0.01, versus medium control.