Anti baffr fitc

PBMCs (1 × 10⁶ cells) were incubated with anti-CD19-PEDy647, anti-CD38-PE, anti-CD41a-FITC, anti-CD41a-PE (Immunotools, Friesoythe, Germany), anti-CD19-PECy7, anti-CD27-APC, anti-BAFFR-FITC, anti-TACI-PE (BioLegend, San Diego, USA), anti-CD86-PE, anti-IgG-PE, anti-IgD-FITC (BD), anti-IgA-FITC, and anti-IgM-PE (Dako, CA, USA) mAbs and the corresponding isotype controls. Whole blood (100 μl) was incubated with anti-CD4-PECy7 (BioLegend), CD5-PE (BD Biosciences), CD19-PEDy647, and CD41a-FITC (Immunotools). Red blood cells were lysed, and white cells fixed using BD FACS lysing solution (BD Biosciences) to be analyzed by flow cytometry.

After pre-incubation with an FcR-blocking reagent, cells were labeled for 30 min on ice with mAb anti-BAFF-R FITC, anti-BCMA PE, and anti TACI APC (Biolegend, San Diego, CA, USA). As positive control for the expression level of BLyS receptors, we used B lymphocytes labeled with mAb anti-CD19 and anti-BLyS receptor mAbs as described above. Appropriate isotype controls were used. Acquisition was performed on a FACS Calibur and included 10,000 events for apoptosis and 50,000 events for BLyS receptors. Data were analyzed using the CellQuest Pro software (BD Immunocytometry Systems; San Jose, CA, USA); the results were expressed as mean fluorescence intensity (MFI).