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Expression and factor-dependent modulation of the interleukin-3 receptor
subunits on human hematopoietic cells
N Sato, C Caux, T Kitamura, Y Watanabe, K Arai, J Banchereau and A Miyajima
Department of Molecular Biology, DNAX Research Institute of Molecular and
Cellular Biology, Palo Alto, CA 94304.
Interleukin-3 (IL-3) regulates growth and differentiation of multipotential
as well as lineage-committed progenitor cells. The human IL-3 receptor
(IL-3R) consists of the alpha and common beta (beta c) subunits. The alpha
subunit (IL-3R alpha) is specific for IL-3 and binds IL-3 with low
affinity. In contrast, the beta c subunit does not bind any cytokine by
itself, but forms a high-affinity receptor with IL- 3R alpha. As the same
beta c subunit also forms high-affinity receptors for IL-5 and
granulocyte-macrophage colony-stimulating factor (GM-CSF) with the
respective cytokine-specific alpha subunit, the expression of the alpha
subunits is responsible for specificity of cytokines. To examine the
expression of IL-3R alpha, we have developed a monoclonal antibody (MoAb),
N3A. N3A specifically bound to cells expressing IL-3R alpha and
immunoprecipitated a 75 Kd glycoprotein, which became 43 Kd on
N-glycosidase digestion. N3A and an anti-beta c antibody, CRS1, were used
in double color fluorescence-activated cell sorter (FACS) staining with
several lineage markers to see the IL-3R expression pattern in peripheral
blood (PB), cord blood (CB), and bone marrow (BM) cells. Both IL-3R
subunits were expressed on myeloid cell lineages (CD13+, CD14+, CD15Lo, or
CD33+). To further study the IL-3R expression on hematopoietic progenitor
cells, the CD34+ populations were isolated from both BM and CB cells. Those
populations showed positive staining profiles with the N3A MoAb and were
weakly stained with the CRS1 MoAb. Furthermore, anti c-kit antibody
staining of the CD34+ fraction from CB, but not from BM, showed two
intensities and the IL-3R alpha expression seemed to be higher in a
fraction of low c-kit expression. Because IL-1, IL-6, G-CSF, stem cell
factor (SCF), interferon (IFN)- gamma, and tumor necrosis factor
(TNF)-alpha are known to enhance IL-3- dependent colony formation, we have
examined whether this enhancement could be correlated with upregulation of
the IL-3R expression. Incubation of CD34+ cells with TNF-alpha for 2 days
significantly increased the level of beta c and G-CSF increased the number
of cells with high level expression of alpha, while other factors did not
affect the IL-3R expression. Thus, different cytokines appear to have
different mechanisms for enhancement of IL-3-dependent proliferation.
Volume 82,
Issue 3,
pp. 752-761,
08/01/1993
Copyright © 1993 by The American Society of Hematology
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