Activation of the erythropoietin (EPO) receptor by bivalent anti-EPO receptor antibodies.
Oligomerization of cytokine receptors including the erythropoietin (EPO) receptor has been advanced as a model for activation. If homodimerization of the EPO receptor activates it, then bivalent antibodies raised to the extracellular domain of the EPO receptor should also homodimerize and activate. Mouse monoclonal antibodies (IgG) raised to the soluble, extracellular domain of the human EPO receptor (EPOR) were found that would stimulate thymidine uptake of an human EPO-dependent cell line, UT-7/EPO. Dose response curves showed bell shapes where activity was low at low and high concentrations. Monovalent (Fab) fragments bound to the receptor but did not stimulate thymidine uptake, which indicates that two antibody binding sites are required for activation. The anti-EPOR antibodies stimulated the formation of burst forming unit erythroid colonies from human CD34(+) cells purified from peripheral blood. This indicates that homodimerization of the EPO receptor by anti-EPOR antibodies is sufficient for both proliferation and differentiation of erythroid progenitor cells and that the constraints on dimerization necessary for activation are rather loose.
Elliott
S
S
Amgen, Amgen Center, Thousand Oaks, California 91320, USA.
Lorenzini
T
T
Yanagihara
D
D
Chang
D
D
Elliott
G
G
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification; D001704:Biopolymers; D002454:Cell Differentiation / Q000276:immunology; D002455:Cell Division / Q000276:immunology; D002460:Cell Line; D015672:Erythroid Precursor Cells / Q000166:cytology; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D017467:Receptors, Erythropoietin / Q000819:agonists* / Q000276:immunology / Q000378:metabolism
D000911:Antibodies, Monoclonal; D001704:Biopolymers; D007140:Immunoglobulin Fab Fragments; D017467:Receptors, Erythropoietin