DCs up-regulate maturation markers in response to stimulation with TLR agonists. Purified mDCs and pDCs were stimulated with the following TLR7 agonists: 10μM R848 and 300 ng of p24^CA equivalents AT-2 HIV for 16-18 hours. Cells were harvested and stained for expression of CCR7, CD40, and CD86. (A) Representative flow cytometric data from primary HIV-infected subject at Fiebig stages I-II. Light gray shading denotes staining of the unstimulated cells with isotype control antibodies. Dark gray shading denotes antibody staining of the unstimulated cells. Solid black lines denote antibody staining of the stimulated cells. Summary of (B) mDC and (C) pDC expression of CCR7, CD40, and CD86 after TLR stimulation. Each bar represents the mean staining (expressed as mean fluorescence intensity; MFI) of DCs from uninfected subjects, subjects at Fiebig stages I-II, subjects at Fiebig stages III-IV, and subjects at Fiebig stages V-VI. Statistical analysis was performed using the nonparametric Mann-Whitney test.