CCL5-mediated chemotaxis of activated CD4⁺ T cells is dependent on PI-3′K and mTOR. (A) Activated peripheral blood (PB) T cells were stained with anti-CCR5 and anti-CD3 antibodies (solid line) or isotype controls (dotted line) and analyzed by FACS. CCL5-mediated chemotaxis is presented as migrated cells per well. (B) Activated PB T cells were pretreated with either DMSO (carrier) or the specified inhibitors for 1 hour at the concentrations indicated, and CCL5-mediated chemotaxis was measured using 10 nM CCL5. The data are presented as percentage migration, with the number of migrated cells at 10 nM CCL5 taken as 100%. Representatives of 3 independent experiments are shown (± SD); *P < .05. (C) Activated PB T cells were pretreated with either ethanol (carrier) or AS-252424 for 1 hour at the concentration indicated, and CCL5-mediated chemotaxis measured using 10 nM CCL5. Data are representative of 2 independent experiments (± SD); *P < .05. (D) Activated PB T cells pretreated with either DMSO (carrier), cycloheximide, or actinomycin D for 30 minutes at the concentrations indicated. The data represent means plus or minus SD of 3 independent experiments. *P < .05.