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Monoclonal antibody 7G3 recognizes the N-terminal domain of the human
interleukin-3 (IL-3) receptor alpha-chain and functions as a specific IL-3
receptor antagonist
Q Sun, JM Woodcock, A Rapoport, FC Stomski, EI Korpelainen, CJ Bagley, GJ Goodall, WB Smith, JR Gamble, MA Vadas and AF Lopez
Division of Human Immunology, Hanson Centre for Cancer Research, Institute
of Medical and Veterinary Science, Adelaide, South Australia.
The human interleukin-3 receptor (IL-3R) is expressed on myeloid, lymphoid,
and vascular endothelial cells, where it transduces IL-3- dependent signals
leading to cell activation. Although IL-3R activation may play a role in
hematopoiesis and immunity, its aberrant expression or excessive
stimulation may contribute to pathologic conditions such as leukemia,
lymphoma, and allergic reactions. We describe here the generation and
characterization of a monoclonal antibody (MoAb), 7G3, which specifically
binds to the IL-3R alpha-chain and completely abolishes its function. MoAb
7G3 immunoprecipitated and recognized in Western blots the IL-3R
alpha-chain expressed by transfected cells and bound to primary cells
expressing IL-3R alpha. MoAb 7G3 bound the IL-3R alpha-chain with a kd of
900 pmol/L and inhibited 125I-IL-3 binding to high- and low-affinity
receptors in a dose-dependent manner. Conversely, IL-3 but not
granulocyte-macrophage colony-stimulating factor (GM-CSF) inhibited
125I-7G3 binding to high- and low-affinity IL- 3Rs, indicating that MoAb
7G3 and IL-3 bind to common or adjacent sites. In keeping with the
inhibition of IL-3 binding, MoAb 7G3 antagonized IL-3 biologic activities,
namely stimulation of TF-1 cell proliferation, basophil histamine release,
and IL-6 and IL-8 secretion from human endothelial cells. Two other
anti-IL-3R alpha-chain MoAbs failed to inhibit IL-3 binding or function.
Epitope mapping experiments using truncated IL-3R alpha-chain mutants and
IL-3R alpha/GM-CSFR alpha chimeras revealed that 31 amino acids in the
N-terminus of IL-3R alpha were required for MoAb 7G3 binding. MoAb 7G3 may
be of clinical significance for antagonizing IL-3 in pathologic conditions
such as some myeloid leukemias, follicular B-cell lymphoma, and allergy.
Furthermore, these results implicate the N-terminal domain of IL-3R alpha
in IL-3 binding. Since this domain is unique to the IL-3/GM- CSF/IL-5
receptor subfamily, it may represent a novel and common binding feature in
these receptors.
Volume 87,
Issue 1,
pp. 83-92,
01/01/1996
Copyright © 1996 by The American Society of Hematology
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