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Identification and analysis of human erythropoietin receptors on a
factor-dependent cell line, TF-1
T Kitamura, A Tojo, T Kuwaki, S Chiba, K Miyazono, A Urabe and F Takaku
Third Department of Internal Medicine, Faculty of Medicine, University of
Tokyo, Japan.
We have recently established a novel cell line, TF-1, from bone marrow
cells of a patient with erythroleukemia, that showed an absolute growth
dependency on each of three hematopoietic growth factors: erythropoietin
(EPO) granulocyte-macrophage colony-stimulating factor (GM-CSF), and
interleukin 3 (IL-3). EPO stimulated the proliferation of TF-1 cells even
at the physiologic concentration (0.03 U/mL). We performed binding
experiments on TF-1 cells using radioiodinated EPO. The binding of
radioiodinated EPO to TF-1 was specific, time- and temperature-dependent,
and saturable. Scatchard analysis of the saturation binding data suggested
the existence of a single class of binding sites (kd = 0.40 nmol/L; number
of binding sites = 1,630 per cell). TF-1 cells were usually maintained in
RPMI 1640 containing 10% fetal bovine serum and 5 ng/mL GM-CSF. The kd and
the number of the EPO receptors were not changed by incubating the cells
with IL-3, although culturing the cells in the presence of EPO resulted in
down-modulation of EPO receptors. The chemical cross-linking study
demonstrated that two molecules with apparent molecular weights of 105
kilodalton (Kd) and 90 Kd were the binding components of EPO. Present data
suggest that human EPO receptors are very similar to the previously
reported murine EPO receptors.
Volume 73,
Issue 2,
pp. 375-380,
02/01/1989
Copyright © 1989 by The American Society of Hematology
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