Phenotype and Function of CD25-Expressing B Lymphocytes Isolated from Human Umbilical Cord Blood
Figure 3
Analysis of surface immunoglobulin expression and secretion from CD25+ and CD25− CD20+ B cells isolated from cord blood and adult blood. In (a) freshly isolated cord blood (N = 10) or adult blood, mononuclear cells (N = 10) were isolated by density gradient centrifugation. Cells were stained for 3-colour flow cytometry analysis using a combination of CD20, CD25, IgG, IgA, IgM, and IgD. In (a), a summary of all 4 surface immunoglobulins expressed by the CD20+CD25+ or CD20+CD25− B cells is shown. Red colour represents IgD, blue colour represents IgM, yellow colour represents IgG, and green colour represents IgA. In (b) and (c), freshly isolated cord blood mononuclear cells (N = 6) were isolated by density gradient centrifugation followed by flow cytometry sorting into CD20+CD25+ or CD20+CD25− populations. These populations were then analysed for the capacity to secret either IgG (b) or IgM (c) by use of ELISPOT. Line in boxes represents median. Statistical evaluation was performed using the parametric unpaired t-test when comparing results from cord blood versus results from adult blood, whereas when comparing results within each group, the paired t-test was used. *, **, and ***.
