tailieunhanh - Báo cáo Y học: Differential response of neuronal cells to a fusion protein of ciliary neurotrophic factor/soluble CNTF-receptor and leukemia inhibitory factor

Department of Physiology, University of Basel, Switzerland; 2Department of Biophysical Chemistry, Biocenter, University of Basel, Switzerland; 3Xerion Pharmaceuticals, Martinsried, Germany; 4Department of Medicine, Section Pathophysiology, Johannes Gutenberg University of Mainz, Germany; 5Department of Biochemistry, Christian Albrechts University of Kiel, Germany Ciliary neurotrophic factor (CNTF) displays neurotrophic activities on motor neurons and neural cell populations both in vivo and in vitro. . | Eur. J. Biochem. 269 3023-3031 2002 FEBS 2002 doi Differential response of neuronal cells to a fusion protein of ciliary neurotrophic factor soluble CNTF-receptor and leukemia inhibitory factor Pia Marz1 Suat Ozbek2 Martina Fischer3 Nicole Voltz4 Uwe Otten1 and Stefan Rose-John4 5 1Department of Physiology University of Basel Switzerland department of Biophysical Chemistry Biocenter University of Basel Switzerland 3Xerion Pharmaceuticals Martinsried Germany department of Medicine Section Pathophysiology Johannes Gutenberg University of Mainz Germany 5Department of Biochemistry Christian Albrechts University of Kiel Germany Ciliary neurotrophic factor CNTF displays neurotrophic activities on motor neurons and neural cell populations both in vivo and in vitro. On target cells lacking intrinsic expression of specific receptor a subunits cytokines of the IL-6 family only act in the presence of their specific agonistic soluble receptors. Here we report the construction and expression of a CNTF soluble CNTF-receptor sCNTF-R fusion protein Hyper-CNTF with enhanced biological activity on cells expressing gp130 and leukemia inhibitory factor receptor LIF-R but not membrane-bound CNTF-R. At the cDNA level the C-terminus of the extracellular domain of human CNTF-R amino acids 1-346 was linked via a single glycine residue to the N-terminus of human CNTF amino acids 1-186 . Recombinant Hyper-CNTF protein was expressed in COS-7 cells. Hyper-CNTF efficiently induced dosedependent STAT3 phosphorylation and proliferation of BAF-3 cells stably transfected with gp130 and LIF-R cDNAs. While on BAF3 gp130 LIF-R cells Hyper-CNTF and LIF exhibited similar biological responses the activity of Hyper-CNTF on pheochromocytoma cells PC12 cells was quite distinct from that of LIF. In contrast to LIF Hyper-CNTF stimulated neurite outgrowth of PC12 cells in a time- and dose-dependent manner correlating with the ability to phosphorylate MAP kinases. These data .

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