Identification of agonistic and antagonistic antibodies against gp190, the leukemia inhibitory factor receptor, reveals distinct roles for its two cytokine-binding domains.
The receptor for the cytokine leukemia inhibitory factor (LIF) associates the low affinity binding component gp190 and the high affinity converter gp130, both of which are members of the family of hematopoietic receptors characterized by the cytokine receptor homology (CRH) domain. The gp190 is among the very few members of this large family to contain two CRH domains. The membrane-distal one (herein called D1) is followed by an Ig-like domain, a membrane-proximal CRH domain called D2, and three type III fibronectin repeats. We raised a series of monoclonal antibodies specific for the human gp190. Among them was the blocking antibody 1C7, which was directed against the D1Ig region and which impaired the binding of LIF to gp190. Another blocking antibody, called 12D3, was directed against domain D2 and interfered with the reconstitution of the high affinity receptor complex, independently of the interaction between LIF and gp190. The blocking effect of these two antibodies concerned four cytokines known to use gp190, i.e. LIF, oncostatin M, ciliary neurotrophic factor, and cardiotrophin-1. Among 23 antibodies tested alone or in combination (two anti-D2 and 21 anti-D1Ig), only the mixture of the two anti-D2 antibodies displayed agonistic activity in the absence of the cytokine. Taken together, these results demonstrate that the two CRH domains of gp190 play different functions in ligand binding and receptor activation.
Taupin
J L
JL
CNRS UMR 5540, Université de Bordeaux II, Bâtiment 1b, 146 rue Léo-Saignat, 33076 Bordeaux Cedex, France. jean-luc.taupin@umr5540.u-bordeaux2.fr
Legembre
P
P
Bitard
J
J
Daburon
S
S
Pitard
V
V
Blanchard
F
F
Duplomb
L
L
Godard
A
A
Jacques
Y
Y
Moreau
J F
JF
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D002455:Cell Division / Q000502:physiology; D002460:Cell Line; D006224:Cricetinae; D016207:Cytokines / Q000378:metabolism* / Q000502:physiology; D000939:Epitopes / Q000276:immunology; D005434:Flow Cytometry; D006801:Humans; D053680:Leukemia Inhibitory Factor Receptor alpha Subunit; D018121:Receptors, Cytokine / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D053606:Receptors, OSM-LIF; D013329:Structure-Activity Relationship
D000911:Antibodies, Monoclonal; D016207:Cytokines; D000939:Epitopes; C507840:LIFR protein, human; D053680:Leukemia Inhibitory Factor Receptor alpha Subunit; D018121:Receptors, Cytokine; D053606:Receptors, OSM-LIF