CMRF-35 cross-linking of pDCs on surface molecule expression. (A) Flow cytometric analysis of HLA-DR and CD80 expression in pDCs cultured with CpG or control ODN after cross-linking with CMRF-35 mAb or left not cross-linked. Each dot represents one analysis sample. MFI is shown. (B) pDCs were cultured with or without CMRF-35 cross-linking in the presence of CpG or control ODN for 24 hours. TNF-α secretion level was measured from the cell culture supernatant. Data are representative of 4 experiments. (C) IL-6 was measured in the cell culture supernatant from pDCs cultured with or without CMRF-35 cross-linking in the presence of CpG or control ODN for 24 hours. Data are representative of 4 experiments. Error bars represent SEM. (D) Flow cytometric analysis of HLA-DR and CD80 expression in pDCs cultured with or without anti–TNF-α antibody and IFN-α for 48 hours. Human IgG1 (hIgG) was used as control antibody for anti–TNF-α. Open histogram with dashed line indicates isotype control; open histogram with solid line (from left to right), no CpG, CpG-hIgG, CpG-anti–TNF-α, CpG-IFN-α; filled histogram (from left to right), CpG + hIgG, CpG + anti–TNF-α, and CpG + IFN-α. Data are from one representative of 3 experiments.
