title: High and low affinity receptors for human interleukin for DA cells/leukemia inhibitory factor on human cells. Molecular characterization and cellular distribution.
abstract: Radioiodinated recombinant human interleukin DA (HILDA)/leukemia inhibitory factor (LIF) purified from conditioned medium of Chinese hamster ovary transfected cells enabled the identification of specific receptor sites on a variety of human cell types. Using low concentrations (up to 500 pM) of the ligand iodinated at a high specific radioactivity, high affinity receptors (equilibrium dissociation constant Kd in the range of 30-100 pM) were first demonstrated. They were expressed at low levels by human peripheral blood monocytes but not by lymphocytes, NK cells, granulocytes, and platelets. The myelomonocytic cell line THP1 as well as the T lymphoma cell line HSB2 and the lymphoblastoid B cell line DAB were also receptor-negative. In contrast, most of the non-lymphoid tumoral cell lines tested, including melanomas, neuroblastomas, and carcinomas, expressed high affinity HILDA/LIF receptors at variable levels (Bmax from 20 to 600 sites/cell). The kinetics of HILDA/LIF high affinity binding to the choriocarcinoma JAR cell line were characterized at 4 degrees C with association and dissociation rate constants of k1 = 2.2 10(9) M-1 min-1 and k-1 = 0.0084 min-1, respectively, corresponding to a steady-state dissociation constant k1/k-1 = 3.8 pM. The subsequent use of higher concentrations of HILDA/LIF labeled at a lower specific radioactivity enabled the identification of a low affinity component on several cell lines (Kd in the range of 1-4 nM; Bmax from 1,000 to 5,000 sites/cell). On JAR cells, this low affinity component was characterized by association and dissociation rate constants at 4 degrees C of k1 = 7.3 10(7) M-1 min-1 and k-1 = 0.19 min-1, respectively (k-1/k1 = 2.6 nM). Affinity cross-linking of HILDA/LIF to JAR cells showed two cross-linked species under both reducing and nonreducing conditions corresponding to receptor species of 120 and 250 kDa, respectively. Whereas both bands had similar intensities under high affinity conditions, the higher band predominated under low affinity conditions. Our data suggest that the 250-kDa chain could constitute the low affinity binding component whereas the association of both 250- and 120-Da subunits would form the high affinity structure.
authors.affiliation: Institut National de la Santé et de la Recherche Médicale (INSERM) U211, Unité de Recherche sur les Effecteurs Lympocytaires T Plateau Technique du CHR, Nantes, France.
authors.forename: A
authors.initials: A
authors.lastname: Godard
authors.forename: D
authors.initials: D
authors.lastname: Heymann
authors.forename: S
authors.initials: S
authors.lastname: Raher
authors.forename: I
authors.initials: I
authors.lastname: Anegon
authors.forename: M A
authors.initials: MA
authors.lastname: Peyrat
authors.forename: B
authors.initials: B
authors.lastname: Le Mauff
authors.forename: E
authors.initials: E
authors.lastname: Mouray
authors.forename: M
authors.initials: M
authors.lastname: Gregoire
authors.forename: K
authors.initials: K
authors.lastname: Virdee
authors.forename: J P
authors.initials: JP
authors.lastname: Soulillou
mesh_terms: D000818:Animals; D016466:CHO Cells; D002460:Cell Line; D006224:Cricetinae; D006131:Growth Inhibitors / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D015850:Interleukin-6; D007700:Kinetics; D053605:Leukemia Inhibitory Factor; D053680:Leukemia Inhibitory Factor Receptor alpha Subunit; D008222:Lymphokines / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D009000:Monocytes / Q000378:metabolism; D018121:Receptors, Cytokine; D011971:Receptors, Immunologic / Q000378:metabolism*; D053606:Receptors, OSM-LIF; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D014162:Transfection; D014407:Tumor Cells, Cultured
chemical_list: D006131:Growth Inhibitors; D015850:Interleukin-6; C507837:LIF protein, human; C507840:LIFR protein, human; D053605:Leukemia Inhibitory Factor; D053680:Leukemia Inhibitory Factor Receptor alpha Subunit; D008222:Lymphokines; D018121:Receptors, Cytokine; D011971:Receptors, Immunologic; D053606:Receptors, OSM-LIF; D011994:Recombinant Proteins