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Prepublished online as a Blood First Edition Paper on January 30, 2003; DOI 10.1182/blood-2002-03-0835.
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Blood, 1 June 2003, Vol. 101, No. 11, pp. 4322-4332
HEMATOPOIESIS
The in vivo profile of transcription factors during neutrophil differentiation in human bone marrow
Malene Digmann Bjerregaard,
Jesper Jurlander,
Pia Klausen,
Niels Borregaard, and
Jack Bernard Cowland
From the the Granulocyte Research Laboratory and the Leukemia and Lymphoma Research Laboratory, Department of Hematology, Rigshospitalet, University of Copenhagen, Copenhagen, Denmark.
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  (C/EBP-) was seen primarily in MBs/PMs, and little expression was found in more mature cells. The level of C/EBP- was constant in the bone marrow–derived cells and decreased in PMNs. C/EBP- was found primarily in MCs/MMs and was almost absent in more mature cells. Expression of C/EBP- , C/EBP- , and C/EBP- 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.
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