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Homodimerization of erythropoietin receptor by a bivalent monoclonal
antibody triggers cell proliferation and differentiation of erythroid
precursors
H Schneider, W Chaovapong, DJ Matthews, C Karkaria, RT Cass, H Zhan, M Boyle, T Lorenzini, SG Elliott and LB Giebel
Department of Receptor Biology, Arris Pharmaceutical Corp, South San
Francisco, CA 94080, USA.
Erythropoietin (EPO) stimulates proliferation and differentiation of
erythroid progenitor cells. Several lines of evidence indicate that the
most likely mechanism of EPO receptor (EPO-R) activation by EPO is
homodimerization of the receptor on the surface of erythrocyte precursors.
Therefore, we argued that it should be possible to raise EPO-R monoclonal
antibodies (MoAbs) that would activate the receptor by dimerization and
thus mimic EPO action. We have identified such an agonist MoAb (MoAb34)
directed against the extracellular EPO binding domain of the EPO-R. This
bivalent IgG antibody triggers the proliferation of EPO-dependent cell
lines and induces differentiation of erythroid precursors in vitro. In
contrast, the monovalent Fab fragment, which cannot dimerize the receptor,
is completely inactive. The mechanism of receptor activation by
homodimerization implies that at high ligand concentrations the formation
of 1:1 receptor/ligand complexes is favored over 2:1 complexes, thereby
turning the ligand agonist into an antagonist. Thus, EPO and MoAb34 should
self-antagonize at high concentrations in both cell proliferation and
differentiation assays. Our data indeed demonstrate that EPO and MoAb34
antagonize ligand-dependent cell proliferation with IC50 values of
approximately 20 and 2 mumol/L, respectively. Erythroid colony formation
(BFUe) is inhibited at MoAb34 concentrations above 1 mumol/L. Furthermore,
we analyzed the MoAb34:EPO-R interaction using a mathematic model
describing antibody-mediated receptor dimerization. The data for
proliferation and differentiation activity were consistent with the
receptor dimer formation on the cell surface predicted by the model.
Volume 89,
Issue 2,
pp. 473-482,
01/15/1997
Copyright © 1997 by The American Society of Hematology
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