Figure 2
Figure 2. Superior IC binding of slanDCs compared with other human blood DCs and monocytes. (A) Measurement of FITC-labeled IC binding (ICs consisted of HSA-FITC and polyclonal IgG anti-FITC) within freshly isolated PBMCs. Binding of ICs to specific cell types was quantified by staining for slanDCs, monocytes, mDCs, and pDCs, followed by flow cytometric analysis. The histograms are a representative example of the IC-binding assay. Light gray histograms show control staining with HSA-FITC. Dark gray histograms show IC binding. The diagram summarizes 5 independent experiments (mean ± SEM). n.d. = not determined. (B) CD16- and CD32-specific IC binding of slanDCs. PBMCs were incubated with 25 µg/mL blocking mAbs for either CD16 (clone 3G8) or CD32 (clone AT10). IC binding of slanDCs in unblocked, and blocked samples were subsequently quantified by flow cytometric analysis. ΔMFI is calculated by subtraction of the MFI of the control staining from the MFI of the samples. (C) Normalized CD16- and CD32-specific IC binding of slanDCs and monocytes. Data are normalized to the HSA-FITC control (0%) and the sample with the control mAb (100%). Data represent mean ± SEM (N = 5).

Superior IC binding of slanDCs compared with other human blood DCs and monocytes. (A) Measurement of FITC-labeled IC binding (ICs consisted of HSA-FITC and polyclonal IgG anti-FITC) within freshly isolated PBMCs. Binding of ICs to specific cell types was quantified by staining for slanDCs, monocytes, mDCs, and pDCs, followed by flow cytometric analysis. The histograms are a representative example of the IC-binding assay. Light gray histograms show control staining with HSA-FITC. Dark gray histograms show IC binding. The diagram summarizes 5 independent experiments (mean ± SEM). n.d. = not determined. (B) CD16- and CD32-specific IC binding of slanDCs. PBMCs were incubated with 25 µg/mL blocking mAbs for either CD16 (clone 3G8) or CD32 (clone AT10). IC binding of slanDCs in unblocked, and blocked samples were subsequently quantified by flow cytometric analysis. ΔMFI is calculated by subtraction of the MFI of the control staining from the MFI of the samples. (C) Normalized CD16- and CD32-specific IC binding of slanDCs and monocytes. Data are normalized to the HSA-FITC control (0%) and the sample with the control mAb (100%). Data represent mean ± SEM (N = 5).

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