Maturation of DCs enhances VLP_HIV-Gag-eGFP and Exosome_DiI capture. (A) Comparative capture of VLP_HIV-Gag-eGFP by DCs. A total of 10⁵ DCs were pulsed for 4 hours at 37°C with 2500 pg p24^Gag in 0.1 mL, washed with PBS, and fixed to analyze the percentage of eGFP-positive cells by FACS. Data show mean values and SEM from 5 independent experiments, including cells from 6 donors. mDCs captured significantly greater amounts of VLPs compared with iDCs (P < .0001, paired t test). (B) Comparative capture of carboxylated yellow fluorescent beads by DCs. A total of 5 × 10⁵ iDCs and mDCs were incubated at 4°C and 37°C for 2 hours with approximately 1.8 × 10¹⁰ beads. Cells were washed, fixed, and analyzed by FACS. Graph displays the percentage of DCs that captured beads at 37°C, after subtracting binding percentages at 4°C. Data show mean values and SEM from 4 independent experiments, including cells from 7 donors. iDCs significantly captured greater amounts of beads compared with mDCs (P = .0042, paired t test). (C) Comparative capture of Jurkat-derived Exosomes_DiI by DCs. A total of 10⁵ DCs were pulsed for 8 hours at 37°C with 150 μg exosomes, washed with PBS, and fixed to analyze the percentage of DiI-positive cells by FACS. Data show mean values and SEM from 4 independent experiments, including cells from 11 donors. mDCs capture significantly greater amounts of exosomes compared with iDCs (P < .0001, paired t test).