Expression of CC-chemokines, CCR5 and chemokine mRNA expression in PBMC cultures after drug treatment.

Panel A: Production of CC-chemokines by PBMC and the effects of adefovir, tenofovir, and PMEO-DAPym on the expression of the CCR5 receptor. Freshly isolated PBMCs from four healthy donors were incubated for 24 h with medium only (NC) or with adefovir, tenofovir, or PMEO-DAPym. We collected the supernatants and measured the concentrations of MIP-1α (▪), MIP-1β (▴), and RANTES (•) using a Bioplex system (Bio-Rad, Hercules, CA). Shown are means ± SEM. The cells were also collected, and the expression of the CCR5 receptor (□) was measured with flow cytometry using the PE-labeled CCR5 (clone 2D7) mAb; it is shown as percentage of control (± SEM) from four independent representative experiments. Panel B: Chemokine mRNA expression upon PMEO-DAPym exposure to PBMC. RT-PCR analysis of MIP-1α, MIP-1β, RANTES, and GAPDH (control) expression in PBMC treated with 100-µg/ml PMEO-DAPym for 4 h. Similar results were obtained with PBMC from three different donors. Results from one representative donor are shown.