Figure 4
Figure 4. Anti-DCIR and BDCA-2 mAbs block binding of HCVcc particles to pDCs. (A) Experimental flowchart. (B) pDCs were pretreated with antibodies against BDCA-2 and DCIR, ILT7, CD123, claudin-1, SR-BI, and CD81, and heparin for 30 minutes at 4°C (full line) or control isotype antibody or heparin control (shadow area). HCVcc particles (MOIHCV RNA = 100) were added to antibody-treated pDCs in the presence of Ca2+ at 4°C, and the binding to pDCs was determined 30 minutes later by anti-E2 mAb conjugated with Alexa-647 and analyzed by flow cytometry. (C-D) Saturating curves of anti-DCIR (C) and anti–BDCA-2 (D) mAbs in pDCs (right ordinate). Relative median fluorescent intensity (MFI) of anti-DCIR and anti–BDCA-2 was normalized to 1 at 0 μg/mL of respective antibodies. Inhibition of HCVcc binding by anti-DCIR (C) and anti–BDCA-2 (D; left ordinate). Representative result of one of 3 experiments is shown. Relative MFI of anti-E2 mAb was normalized to 1 at 30 μg/mL of anti-DCIR and anti–BDCA-2.

Anti-DCIR and BDCA-2 mAbs block binding of HCVcc particles to pDCs. (A) Experimental flowchart. (B) pDCs were pretreated with antibodies against BDCA-2 and DCIR, ILT7, CD123, claudin-1, SR-BI, and CD81, and heparin for 30 minutes at 4°C (full line) or control isotype antibody or heparin control (shadow area). HCVcc particles (MOIHCV RNA = 100) were added to antibody-treated pDCs in the presence of Ca2+ at 4°C, and the binding to pDCs was determined 30 minutes later by anti-E2 mAb conjugated with Alexa-647 and analyzed by flow cytometry. (C-D) Saturating curves of anti-DCIR (C) and anti–BDCA-2 (D) mAbs in pDCs (right ordinate). Relative median fluorescent intensity (MFI) of anti-DCIR and anti–BDCA-2 was normalized to 1 at 0 μg/mL of respective antibodies. Inhibition of HCVcc binding by anti-DCIR (C) and anti–BDCA-2 (D; left ordinate). Representative result of one of 3 experiments is shown. Relative MFI of anti-E2 mAb was normalized to 1 at 30 μg/mL of anti-DCIR and anti–BDCA-2.

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