Surface expression of CD300a/c on pDCs and their down-regulation by R837 and CpG. (A) Mononuclear cells from peripheral blood and tonsil tissue were isolated using Ficoll-Paque Plus density centrifugation. Cells were stained with CMRF-35 mAb, anti–mouse Ig F(ab2)′-PE, and BDCA-4-APC, or appropriate isotype controls, and analyzed by flow cytometry. Data are representative of 8 blood and 4 tonsil samples. (B) Blood pDCs were stimulated for 48 hours with R837 or CpG. pDCs were treated with chloroquine (CQ; 10 μM) for 30 minutes before stimulation with CpG. The histograms show cells stained with CMRF-35 or isotype control, to monitor the expression of CD300a/c by flow cytometry. Open dashed line indicate isotype control; open solid line, no stimuli; gray filled histogram, with R837; black filled histogram, with CpG; filled with slash line, CQ + CpG. Data are representative of 3 experiments.
