Enrichment and characterization of uncommitted B-cell precursors from fetal liver at day 12 of gestation.

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RESUMO

We describe an assay system that allows precursor cells, uncommitted for heavy and light chain immunoglobulin expression, to develop into B lymphocytes that can differentiate to antibody-producing cells. Some precursors have the immunoglobulin loci in germ-line configuration. Approximately 200-1500 precursor cells are present in one fetal liver by day 12 of gestation; they express the surface marker AA4.1. Most precursors do not express the B220 marker. Commitment to heavy chain immunoglobulin expression occurs after an average of two cell division; commitment to light chain expression takes place after two additional rounds of division. DNA analysis from the progeny of single precursor cells shows that: (i) most B220- precursor cells have not completed D-J rearrangement (9/11) and some were in germ line configuration (4/11); and (ii) most B220+ precursor cells exhibit two D-J rearrangements (4/5 samples). These experiments define two types of B-lymphocyte precursor cells in fetal liver: the first, B220+ AA4.1+, acquires the capacity to respond to mitogens only after 5 days in culture, and does not have productive V-D-J rearrangements but might exhibit two stable D-J rearrangements; the second, B220- AA4.1+, acquires the capacity to respond to mitogens only after 9 days in culture and can be in germ-line configuration in the Ig loci, and undergoes rearrangement of heavy and light chain genes in vitro. Both precursor types require interaction with stromal cells before becoming responsive to interleukin 7.

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