Prospective isolation and global gene expression analysis of the erythrocyte colony-forming unit (CFU-E)

The erythrocyte colony-forming unit (CFU-E) is a rare bone marrow (BM) progenitor that generates erythrocyte colonies in 48 hours. The existence of CFU-Es is based on these colonies, but CFU-Es have not been purified prospectively by phenotype. We have separated the "nonstem," "nonlymphold" compartment (lineage marker [lin](-)c-Kit(+)Sca-1(-)IL-7Ralpha(-)) into interleukin 3 receptor alpha negative (IL-3alpha(-)) and IL-3Ralpha(+) subsets. Within IL-3Ralpha(-) but not lL-3Ralpha(+) cells we have identified TER119(-)CD41(-)CD71(+) erythrocyte-committed progenitors (EPs). EPs generate CFU-E colonies at about 70% efficiency and generate reticulocytes in vivo. Depletion of EPs from BM strongly reduces CFU-E frequencies. EPs lack potential for erythrocyte burst-forming unit, megakaryocyte, granulocyte (G), and monocyte (M) colonies, and for spleen colony-forming units. Chronically suppressed erythropolesis in interferon consensus sequence-binding protein (ICSBP)deficient BM is associated with reduced frequencies of both the EP population and CFU-E colonies. During phenylhydrazine-induced acute anemia, numbers of both the EP population and CFU-E colonies increase. Collectively, EPs(lin(-)c-KIt(+)Sca-1(-)IL-7Ralpha(-)IL-3Ralpha(-)CD41(-)CD71(+) ) account for most, if not all, CFU-E activity in BM. As a first molecular characterization, we have compared global gene expression in EPs and nonerythroid GM progenitors. These analyses define an erythrold progenitor-specific gene expression pattern. The prospective isolation of EPs is an important step to analyze physiologic and pathologic erythropoiesis.