ABSTRACTS – ORAL PRESENTATIONS - AMCA, spol. s r.o.

P68. SUBPOPULATIONS OF B LYMPHOCYTES - BASIC FLOW CYTOMETRIC ANALYSIS
Vlas T., Gutová V., Panzner P.
Department of Immunology and Allergology, Charles University in Prague, Faculty of
Medicine and University Hospital in Pilsen, Pilsen, Czech Republic; vlast@fnplzen.cz
B lymphocytes are an important group of white blood cells. B lymphocytes in their lives
go through many changes. These changes are reflected by different expression of CD
markers on the cell membrane. Presence of different CD markers divides B lymphocytes
into several groups. Most frequent diseases with changes in subpopulations of B cells
are common variable immunodeficiency (CVID) and selective IgA deficiency (IGAD).
Changes in subpopulations are important in CVID for Freiburg classification. Detailed
analysis of B lymphocyte abnormalities, and their interrelations and association with
clinical symptoms could contribute to clarifying the causes of these diseases.
Material and Methods:
We analyzed 40 samples of heparinised blood. Samples for analysis were separated
on a density gradient (Histopaque 1077, SigmaAldrich, UK). Using flow cytometry we
determined the percentage of cells expressing different CD molecules (CD19, CD21,
CD27, CD38, IgM and IgD). Monoclonal antibodies used Anti-HU CD19 PE-DyLight 594,
Anti-HU CD21 FITC, Anti-HU CD27 PE-Cy5, Anti-HU CD38 PE-Cy5, Anti-HU IgM PE (all from
Exbio, CZE) and Anti-HU IgD FITC (Beckmann Coulter, USA).The analysis was performed on
flow cytometer FC500 (Beckmann Clouter, USA) with software CXP analysis (Beckmann
Coulter, USA).
Results:
We found a significant increase by 26.4% (p