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Expression and extracellular release of transferrin receptors during
peripheral erythroid progenitor cell differentiation in liquid culture
N Shintani, Y Kohgo, J Kato, H Kondo, K Fujikawa, E Miyazaki and Y Niitsu
Department of Internal Medicine, Sapporo Medical University School of
Medicine, Japan.
The expression and extracellular release of transferrin receptor (TR) was
investigated by in vitro model system of erythroid differentiation. Human
peripheral blood mononuclear cells were cultured with interleukin- 3 (IL-3)
for 7 days, and with erythropoietin (EPO) for an additional 8 days. After
EPO stimulation, IL-3-stimulated blastic cells were serially differentiated
into mature erythrocytes. [3H]-thymidine incorporation of cultured cells
increased linearly from day 0 to 5, followed by a decrease. Flow cytometric
analysis showed an increase of TR expression from day 0 to 5, followed by a
slight decrease. By metabolic labeling with [35S]methionine and
immunoprecipitation, the cell lysate exhibited a 95-kD band corresponding
to the intact TR on sodium dodecyl sulfate-polyacrylamide gel
electrophoresis/autoradiography at day 5, when polychromatic erythroblasts
had their peak. The culture supernatant solubilized by tween-20 exhibited a
95-kD and an 85-kD band on days 5 and 8, which corresponded to the intact
and the truncated forms of TR, respectively. The 95-kD band was more
intense at day 5 than at day 8. The reverse transcriptase-polymerase chain
reaction assay showed that the receptor- mRNA expression was parallel to
receptor synthesis. Thus, the synthesis and expression of TR on
erythrocytes is associated mainly with cell proliferation in the early
phase, and with both cell proliferation and hemoglobin production in the
middle to late phases of maturation. Concomitantly, the extracellular
release of TR from erythrocytes occurs in the middle to late phases of
maturation. These data suggest that polychromatic erythroblasts release
soluble TR as both intact and truncated forms and may be an important
source of serum TR implicated as an index for erythropoietic activity in
the marrow.
Volume 83,
Issue 5,
pp. 1209-1215,
03/01/1994
Copyright © 1994 by The American Society of Hematology
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