Alteration of LPS-stimulated DC chemotaxis, calcium mobilization, and chemokine receptor expression by TPT treatment. (A) Effects of TPT treatment on DCs and LPS-stimulated DC chemotactic response to RANTES and SLC. Chemokine (RANTES and SLC) was placed in the lower well of chemotaxis chamber. DC suspension was placed in the upper well of the chemotaxis chamber. Polycarbonate filters separated the upper and lower wells. After incubation, the cells that migrated across the filters were stained and counted. The open bars show data from untreated control group; black bars, data from the TPT (D2-7)–treated group. Comparison with untreated control: ^*P < .05; ^**P < .01; ^***P < .001. Error bars indicate SD. (B) Effects of TPT (D2-7) treatment on RANTES- or SLC-mediated calcium mobilization by DCs and LPS-stimulated DCs. The cultured DCs and LPS-stimulated DCs were loaded with Furo-2 and stimulated with 500 ng/mL of RANTES or SLC. The ratio of fluorescence at 340- and 380-nm wavelengths was recorded and calculated using FL Win Lab program (Perkin-Elmer Life and Analytical Sciences). Arrows indicate addition of stimuli. (C) Effects of TPT (D2-7) treatment on CCR5 and CCR7 expression by Day 9 LPS-stimulated DCs. For cells labeled with mAb or isotype IgG, the CCR5 or CCR7 expression was analyzed by FACS. Dashed-line histogram shows isotype control; gray histogram shows untreated control of LPS-stimulated DCs, and solid-line histogram shows LPS-stimulated DCs which were pretreated with TPT(D2-7). The data are representative of 3 separate experiments with similar results.