Loss of c-kit accompanies B-lineage commitment and acquisition of CD45R by most murine B-lymphocyte precursors.
Using surface markers, we identified two bone marrow (BM) subsets enriched for TdT+ cells on the brink of CD45R acquisition. These two populations, Lin-c-kitLo and Lin-c-kit-, consisting of 35.4% and 7. 4%, respectively, TdT+ cells, generated B-lineage cells in overnight cultures. Approximately half of the c-kitLo B-lineage precursors were bipotential, yielding myeloid and lymphoid progeny, whereas most that were c-kit- gave rise only to lymphocytes. Analysis of B-lineage progression during a finite culture period showed that the most mature precursors were concentrated in the Lin-c-kit- population. Moreover, a majority of the earliest CD45R+ pro-B cells in BM, identified as CD45R+ CD43(+) BP-1(-) CD25(-) natural killer (NK)1.1(-) sIgM-, were also c-kit-. These c-kit- cells, like their c-kitLo counterparts, expressed TdT, proliferated in response to interleukin (IL)-7, and generated sIgM+ cells. These data suggest that TdT expression is initiated as c-kit downregulation begins in Lin- cells, with progressive loss of c-kit during B-lineage differentiation. CD45R expression is initiated during the transition from c-kitLo to c-kit- with many cells losing c-kit before acquiring CD45R. The ability to isolate highly enriched populations of viable CD45R- precursors will be instrumental in characterizing the earliest B-lineage cells.
Oklahoma Medical Research Foundation, Immunobiology and Cancer Program, Oklahoma City, OK, USA.
K J
KJ
Payne
K L
KL
Medina
P W
PW
Kincade
D000818:Animals; D001402:B-Lymphocytes / Q000166:cytology* / Q000378:metabolism; D015415:Biomarkers; D001854:Bone Marrow Cells / Q000166:cytology; D002454:Cell Differentiation / Q000235:genetics; D019070:Cell Lineage; D004253:DNA Nucleotidylexotransferase / Q000032:analysis; D005260:Female; D018507:Gene Expression Regulation, Developmental; D006410:Hematopoiesis / Q000235:genetics*; D017493:Leukocyte Common Antigens / Q000096:biosynthesis / Q000235:genetics; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D019009:Proto-Oncogene Proteins c-kit / Q000096:biosynthesis* / Q000235:genetics
D015415:Biomarkers; D019009:Proto-Oncogene Proteins c-kit; D004253:DNA Nucleotidylexotransferase; D017493:Leukocyte Common Antigens