Characterization of B-Lymphocytes

Interleukin-21 restores immunoglobulin production ex vivo in patients with common variable immunodeficiency and selective IgA deficiency

“…29 Three cocktails of monoclonal antibodies were used to determine B-cell and plasma cell populations: (1) anti-CD19-Rphycoerythrin-Cyanine 7 (PC7), anti-CD27-fluorescein isothiocyanate (FITC; both from DAKO), anti-CD5-allophycocyanin (APC; BD Biosciences), and anti-IgD-phycoerythrin (PE) (SouthernBiotech); (2) anti-CD19-PC7, anti-IgD-PE, anti-CD38-APC, and anti-CD77-FITC (both from BD Biosciences); (3) anti-CD19-PC7, anti-CD138-PE (Miltenyi Biotec), and anti-IgA-FITC and anti-IgG-APC (both from Jackson ImmunoResearch) or anti-CD38-APC. All antibodies were initially pretitrated to ensure uniform compensation characteristics, and isotype controls (mouse IgG1, rabbit F(abЈ) 2 , goat F(abЈ) 2 ; all from Jackson ImmunoResearch) were embedded to detect unspecific staining.…”

T and B lymphocyte abnormalities in bone marrow biopsies of common variable immunodeficiency

“…Twenty-five biopsies were accompanied by an analysis of early B-cell differentiation markers in BM aspirates using flow cytometry. 20 Whereas the hematopoietic compartment of the 47 evaluable BM biopsies was inconspicuous, the key findings relate to the T-and B-cell compartment of the BM. First, in line with the pronounced hypogammaglobulinemia, BM plasma cells were either absent or greatly reduced in 94% of the biopsies confirming a previous study.…”

“…The protocol for analyzing B-cell precursors is essentially based on the work of the van Dongen group 19 and has been described in detail. 20 Four-color flow cytometric analysis (FACSCalibur; BD Biosciences) was performed using FITC, phycoerythrin (PE), peridinin chlorophyll protein (PerCP), phycoerythrincyanin 7, or allophycocyanin-conjugated antibodies: CD3 (clone SK7, PerCP, or allophycocyanin), CD19 (clone SJ25C1, allophycocyanin), CD20 (clone Leu-16, PE), CD33 (clone P67.6; PerCP), CD34 (clone 8G12, FITC), CD45 (clone 2D1, PerCP) by BD Biosciences; CD10 (clone ALB1, FITC), CD16 (clone 3G8, phycoerythrin-cyanin 7), CD19 (clone J4.119, FITC, or phycoerythrin-cyanin 7), CD22 (clone SJ10.1H11, PE), CD24 (clone ALB9, PE), CD36 (clone FA6.152, FITC), CD79a (HM47, PE), CD179a (VpreB, clone 4G7, PE) by Beckman Coulter; TdT (clone HT-6; FITC), anti-IgD and anti-IgM [rabbit F(abЈ) 2 , FITC] by Dako Denmark; anti-IgD and anti-IgM [goat F(abЈ) 2 , PE] by Southern Biotechnology. For intracellular staining, the IntraPrep kit (Beckman Coulter) was used.…”