The status of Ig loci rearrangements in single cells from different stages of B cell development

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Abstract
Differential expression of c-kit, CD25 (TAC), surrogate L chain and cytoplasmic μH chain, and surface expression of IgM and IgD allows the separation of B220 (CD45+)B cell subpopulations. PCR analyses with DNA of single cells developed by others and by us have been used to monitor the conformation of the Ig H and L chain gene loci in these different B lineage subpopulations. The results of these analyses indicate that B220+/c-kit+/CD25− cells are the precursors of large B220+/CD25+/slgM− which, in turn, are the precursors of small B220+/CD25+/slgM− cells. The majority of B220+/c-kit+/CD25− cells are DHJH-rearranged, with L chain loci in germline configuration and are thus pre-B I cells. More than 90% of all large B220+/CD25+/slgM− cells have at least one H chain locus VHDHJH rearranged; half of them have also the second locus VHDHJH rearranged and are thus large pre-B II cells. Rearrangements of at least one allele of the kL chain loci become detectable in 65% of the small B220+/CD25+/slgM− cells, 67% of the immature B and >75% of the mature B cells. The ratio of kL to λL gene rearrangements in all three subpopulations is ˜10:1, indicating that the kL/λL ratio is established as soon as rearrangements are made.

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