Anti cd38 pe

Fresh leukapheresis products were stained with the following monoclonal antibodies (MoAbs): anti-CXCR4–PE, anti-CD49d–PE, anti-CD11a–PE, anti-CD34–FITC (BD Biosciences, Franklin Lakes, NJ) or corresponding isotype controls. Additionally, PB mononuclear cells (PBMNCs) were isolated by density gradient centrifugation, viably frozen, and stored in liquid nitrogen until use. Thawed PBMNCs were stained with anti-CD34–FITC (BD Biosciences), CD44–FITC, anti-CD38–PE, anti-CD34–PE (Beckman Coulter), and anti-CD133–APC MoAbs (Miltenyi Biotec, Bergisch-Gladbach, Germany). The percentage of cells expressing respective receptors and mean fluorescence intensity values in relation to isotype controls (MFIRs) were analyzed with a flow cytometer and its accompanying software (Gallios and Caluza, respectively, both Beckman Coulter).

EDTA whole blood samples from 5 healthy subjects, 10 patients with mild and 10 patients with severe disease were stained with anti-HLA-DR PE and ECD, anti-CD3 ECD, anti-CD4 PE, anti-CD8 FITC, anti-CD19 PC7, anti-CD14 FITC, anti-CD16 PC5, anti-CD15 PE, anti-CD57 FITC, anti-CD56 PE, anti-CD11c PCP, anti-CD123 PE, anti-CD83 PE, anti-CD38 PE, anti-CD23 ECD and isotype controls (all from Beckman Coulter) for 20 minutes in the dark at +4°C. Samples were analyzed on the flow cytometer Cytomics FC500 (Beckman Coulter). Data were processed by FlowJo V.10.