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Originally published online as doi:10.1189/jlb.1003474 on December 23, 2003

Published online before print December 23, 2003
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(Journal of Leukocyte Biology. 2004;75:569-578.)
© 2004 by Society for Leukocyte Biology

End-stage differentiation of neutrophil granulocytes in vivo is accompanied by up-regulation of p27kip1 and down-regulation of CDK2, CDK4, and CDK6

Pia Klausen1, Malene Digmann Bjerregaard, Niels Borregaard and Jack Bernard Cowland

The Granulocyte Research Laboratory, Department of Hematology, Rigshospitalet, University of Copenhagen, Denmark

1 Correspondence: The Granulocyte Research Laboratory, Rigshospitalet, Department of Hematology 93.2.2, 20 Juliane Mariesvej, DK-2100 Copenhagen, Denmark. E-mail: pklausen{at}rh.dk

The in vivo expression profiles of cell-cycle proteins regulating G1-to-S-phase transition were determined in three neutrophil precursor populations from human bone marrow: myeloblasts (MBs) and promyelocytes (PMs); myelocytes (MCs) and metamyelocytes (MMs); and band cells (BCs) and segmented neutrophil cells (SCs) and in mature polymorphonuclear neutrophils (PMNs) from peripheral blood. Complete cell-cycle arrest was observed in BCs/SCs and PMNs. Cyclins D1, D2, and D3 were found to be down-regulated during granulopoiesis, whereas a slight increase of cyclin E was seen. In contrast, cyclin-dependent kinase (CDK)2, -4, and -6 were down-regulated from the MC/MM stages and onward. The transcript levels of CDK2, -4, and -6 were concurrently down-regulated. As the only CDK inhibitor, p27kip1 protein and mRNA expression were up-regulated in MCs/MMs and reached peak levels in PMNs. Protein expression of retinoblastoma protein and the related pocket proteins p107 and p130 was down-regulated from the MC/MM stages and onward. This is the first report to describe expression levels of cell-cycle proteins during granulopoiesis in vivo, and it strongly contrasts the observations made in cell-culture systems in vitro.

Key Words: granulopoiesis • cell cycle • bone marrow




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