title: The in vivo profile of transcription factors during neutrophil differentiation in human bone marrow.
abstract: In vivo distribution of myeloid transcription factors during granulopoiesis was investigated by Northern and Western blotting in 3 neutrophil precursor populations from human bone marrow: immature (myeloblasts [MBs] and promyelocytes [PMs]); intermediate mature (myelocytes [MCs] and metamyelocytes [MMs]); and mature neutrophil cells (band cells [BCs] and segmented neutrophil cells [SCs]). Nonneutrophil cells were removed with magnetic-bead-coupled antibodies against CD2, CD3, CD14, CD19, CD56, CD61, glycophorin-A, and CD49d (BCs/SCs) before RNA and protein extraction. Polymorphonuclear neutrophils (PMNs) from peripheral blood depleted with anti-CD49d antibodies were also included. Expression of acute myeloid leukemia 1b (AML-1b), c-myb, GATA-1, and CCAAT/enhancer binding protein gamma (C/EBP-gamma) was seen primarily in MBs/PMs, and little expression was found in more mature cells. The level of C/EBP-alpha was constant in the bone marrow-derived cells and decreased in PMNs. C/EBP-epsilon was found primarily in MCs/MMs and was almost absent in more mature cells. Expression of C/EBP-beta, C/EBP-delta, and C/EBP-zeta was observed from the MC/MM stage onward, with peak levels in the most mature cells. The amount of PU.1 increased throughout maturation whereas the level of Elf-1 reached a nadir in MCs/MMs The PU.1 coactivator c-jun and c-jun's dimerization partner c-fos were both detectable in MCs/MMs and increased in amount with maturity. CCAAT displacement protein (CDP) was found at comparable levels at all stages of differentiation. This demonstrates a highly individualized expression of the transcription factors, which can form the basis for the heterogeneous expression of granule proteins during granulopoiesis and cell cycle arrest in metamyelocytes.
authors.affiliation: Department of Hematology, Rigshospitalet, University of Copenhagen, Denmark. malene.bjerregaard@rh.dk
authors.forename: Malene Digmann
authors.initials: MD
authors.lastname: Bjerregaard
authors.forename: Jesper
authors.initials: J
authors.lastname: Jurlander
authors.forename: Pia
authors.initials: P
authors.lastname: Klausen
authors.forename: Niels
authors.initials: N
authors.lastname: Borregaard
authors.forename: Jack Bernard
authors.initials: JB
authors.lastname: Cowland
mesh_terms: D015703:Antigens, CD / Q000032:analysis; D015152:Blotting, Northern; D015153:Blotting, Western; D001854:Bone Marrow Cells / Q000166:cytology* / Q000378:metabolism; D002454:Cell Differentiation / Q000235:genetics; D006098:Granulocytes / Q000166:cytology; D006801:Humans; D019891:Leukopoiesis / Q000235:genetics; D009504:Neutrophils / Q000166:cytology / Q000378:metabolism*; D012333:RNA, Messenger / Q000032:analysis; D014157:Transcription Factors / Q000032:analysis* / Q000235:genetics
chemical_list: D015703:Antigens, CD; D012333:RNA, Messenger; D014157:Transcription Factors