Ordering of human bone marrow B lymphocyte precursors by single-cell polymerase chain reaction analyses of the rearrangement status of the immunoglobulin H and L chain gene loci

CD19(+)CD10(+) human B lineage bone marrow cells were separated into cycling or resting cells, which differ in their expression of CD34, V-preB, recombination activating gene (RAG-1), and terminal deoxynucleotidyl transferase (TdT). Polymerase chain reaction analyses developed for D(H)J(H) and V(kappa)J(kappa), V(kappa)J(kappa)K((de)) and VkappaK(de) rearrangements with DNA of single cells and a comparison with B lineage cell development in mouse bone marrow, allow to delineate the human B lymphocyte pathway of development as follows: CD34(+)V(preB)(+)RAG-1(+)TdT(+), D(H)J(H)-rearranged, kappa L germline cycling pre-B I cells --> CD34(-)V(preB)(+)mu H chain(+) (pre-B receptor(+)) RAG-1(-)TdT(-), V(H)D(H)J(H)-rearranged, kappa L germline, cycling pre-B II cells --> CD34(-)V(preB)(-), intracytoplasmic mu H chain(+) (pre-B receptor(-)) RAG-1(+/-) TdT(-), V(H)D(H)J(H)-rearranged, mainly kappa L germline cycling pre-B II cells --> CD34(-)V(preB)(-) intracytoplasmic mu H chain(+), RAG-1(+)TdT(-), V(H)D(H)J(H)(-)rearranged, V(kappa)J(kappa)-rearranged IgM(-), resting pre-B II cells CD34(+)V(preB)(-), sIgM(+), RAg-1(+)TdT(-), V(H)D(H)J(H)- and V(kappa)J(kappa)-rearranged IgM(+) immature B cells --> Cd34(-), CD10(-), sIgM(+)/sIgD(+) mature B cells. This order, for the first time established for human B lineage cells, shows striking similarities with that established for mouse B lineage cells in bone marrow.