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," "nonlymphoid" compartment (lineage marker [lin]-c-Kit+Sca-1-IL-7Ralpha-) into interleukin 3 receptor alpha negative (IL-3Ralpha-) and IL-3Ralpha+ subsets. Within IL-3Ralpha- but not IL-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 erythropoiesis 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 erythroid progenitor-specific gene expression pattern. The prospective isolation of EPs is an important step to analyze physiologic and pathologic erythropoiesis.
Terszowski
Grzegorz
G
Department for Immunology, University of Ulm, D-89081 Ulm, Germany.
Waskow
Claudia
C
Conradt
Peter
P
Lenze
Dido
D
Koenigsmann
Jessica
J
Carstanjen
Dirk
D
Horak
Ivan
I
Rodewald
Hans-Reimer
HR
D000740:Anemia / Q000139:chemically induced / Q000473:pathology; D000818:Animals; D001854:Bone Marrow Cells; D019070:Cell Lineage; D002469:Cell Separation; D004195:Disease Models, Animal; D015672:Erythroid Precursor Cells / Q000166:cytology* / Q000378:metabolism; D004920:Erythropoiesis / Q000235:genetics; D020869:Gene Expression Profiling; D016130:Immunophenotyping; D050835:Interferon Regulatory Factors; D051379:Mice; D008810:Mice, Inbred C57BL; D010659:Phenylhydrazines; D012313:RNA / Q000032:analysis; D012097:Repressor Proteins; D000099185:Interferon Regulatory Factor-8
D050835:Interferon Regulatory Factors; D010659:Phenylhydrazines; D012313:RNA; D012097:Repressor Proteins; D000099185:Interferon Regulatory Factor-8; C030299:phenylhydrazine