Journal
SCANDINAVIAN JOURNAL OF IMMUNOLOGY
Volume 62, Issue -, Pages 67-72Publisher
BLACKWELL PUBLISHING
DOI: 10.1111/j.1365-3083.2005.01612.x
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The in vitro differentiation of B-lineage progenitors into Ig-secreting mature B cells has classically required a co-culture system containing lipopolysaccharide (LPS) and stromal cells. We have previously showed that B-lineage progenitors cultured in round-bottomed wells can mature and secrete immunoglobulin M (IgM) on par with cultures containing stromal cells. This clearly demonstrates that any factors essential for progenitor cell maturation can be found in cultures containing media, serum, LPS and B-cell progenitors. However, stromal cells are important for the maturation observed when cells are cultured in flat-bottomed wells. We hypothesized that stromal cells may attract B-cell progenitors and promote contacts between responsive cells, a phenomenon that is mimicked by the cultures in round-bottomed wells. In this study, we explore how stromal cells accomplish these functions. We show that stromal cells attract B-cell progenitors in a pertussis toxin-sensitive manner. The stromal cell line S17 produces the chemokine CXCL12, which is able to induce the chemotaxis of B-lineage progenitors. Chemotaxis can be blocked by a small peptide inhibitor (T134) of CXCR4, the CXCL12 receptor. Further, disrupting chemotaxis can reduce the supportive role played by S17 when B-lineage progenitors are cultured in flat-bottomed wells.
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