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Changing the differentiation program of hematopoietic cells: retinoic
acid-induced shift of eosinophil-committed cells to neutrophils
CC Paul, S Mahrer, M Tolbert, BL Elbert, I Wong, SJ Ackerman and MA Baumann
Research Service, VA Medical Center, Dayton, OH 45428, USA.
The mechanisms by which hematopoietic progenitor cells become lineage-
committed remain poorly understood. A cloned subline of the AML14 cell line
(AML14.3D10) that spontaneously differentiates to eosinophilic myelocytes
in the absence of cytokine stimulation was obtained by limiting dilution.
This subline exhibits augmented expression of interleukin-5 (IL-5) receptor
alpha subunit mRNA and synthesizes all major eosinophil granule proteins.
Exposure of this cell line to all- trans retinoic acid (ATRA) causes loss
of eosinophilic granules and fast green staining within 48 hours, without
cell death. In addition, mRNA for the IL-5 receptor alpha subunit becomes
undetectable by 48 hours and the cells lose responsiveness to IL-5. Major
basic protein, measured as a marker of eosinophilic granule content,
decreases from more than 16 pg/cell to undetectable levels by 5 days after
ATRA. Concomitant with the loss of major basic protein and fast green
staining, surface expression of CD16 becomes detectable and is maximum by
10 days after ATRA. mRNA for the granulocyte colony-stimulating factor
(G-CSF) receptor becomes detectable by day 5, and the cells become
responsive to G-CSF. At this time, the cells appear morphologically as
mature neutrophils and can reduce nitroblue tetrazolium. With continued
culture, the neutrophilic cells die and the culture becomes repopulated
with eosinophilic myelocytes. These findings show that it is possible to
change the differentiation program of hematopoietic cells even after they
show evidence of advanced lineage commitment. The AML14.3D10 subclone of
AML14 will be a valuable model for study of the transcriptional regulation
of the eosinophil and neutrophil differentiation programs and
lineage-specific gene expression.
Volume 86,
Issue 10,
pp. 3737-3744,
11/15/1995
Copyright © 1995 by The American Society of Hematology
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