Flow Cytometry Assessment of In Vitro Generated CD138+ Human Plasma Cells
Figure 2
DL-101 binding on human cells lines, blood CD19+ cells, and in vitro plasma cells. RPMI-8226, SKW6.4, and Ramos cell lines were mixed in equal proportion and stained with PE-conjugated B-A38 and DL-101 alone or combined (MIX). The staining profiles are presented for the three conditions (a). The same three anti-CD138 conditions were also used on four independent samples of blood mononuclear cells. (b) Sytox Blue, CD45, and CD19 were first used to gate the viable CD19+CD45+ cells which were then analyzed for CD138 expression. (c) Differentiated B lymphocytes, prepared as in Figure 1, were simultaneously stained with PerCP-conjugated CD19 antibodies, APC-efluor-conjugated B-A38, and either PE-conjugated B-B4 or DL-101. Analyses were done on gated CD19+ viable cells. An example representative of 5 independent experiments is shown. R1 gate included double-positive cells for B-A38 (BA) and B-B4 (BB) mAbs while R2 and R3 gates included single-positive cells for B-A38 (BA) or DL-101 (DL) mAbs, respectively. (d) The gates established in (c), namely, BA+BB+ (R1), (R2), and (R3), were used to establish the proportion of these CD138+ cells subsets for five independent B lymphocyte cultures. The results are shown as the mean ± S.E.M. No significant differences were observed between all possible comparisons (Tukey-Kramer multiple comparisons test, ).
