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Development of an interleukin (IL) 6 receptor antagonist that inhibits IL-6-dependent growth of human myeloma cells
- Source :
- The Journal of Experimental Medicine
- Publication Year :
- 1994
-
Abstract
- The pleiotropic cytokine interleukin 6 (IL-6) plays a role in the pathogenesis of various diseases, such as multiple myeloma, autoimmune and inflammatory diseases and osteoporosis. Therefore, specific inhibitors of IL-6 may have clinical applications. We previously succeeded in developing receptor antagonists of IL-6 that antagonized wild-type IL-6 activity on the human Epstein-Barr virus (EBV)-transformed B cell line CESS and the human hepatoma cell line HepG2. However, these proteins still had agonistic activity on the human myeloma cell line XG-1. We here report the construction of a novel mutant protein of IL-6 in which two different mutations are combined that individually disrupt the association of the IL-6/IL-6 receptor (R) alpha complex with the signaltransducing "beta" chain, gp130, but leave the binding of IL-6 to IL-6R alpha intact. The resulting mutant protein (with substitutions of residues Gln160 to Glu, Thr163 to Pro, and replacement of human residues Lys42-Ala57 with the corresponding residues of mouse IL-6) was inactive on XG-1 cells and weakly antagonized wild-type IL-6 activity on these cells. By introducing two additional substitutions (Phe171Leu, Ser177Arg), the affinity of the mutant protein for IL-6R alpha was increased fivefold, rendering it capable of completely inhibiting wild-type IL-6 activity on XG-1 cells. Moreover, this mutant also antagonized the activity of IL-6, but not that of leukemia inhibitory factor, oncostatin M, or GM-CSF on the human erythroleukemia cell line TF-1, demonstrating its specificity for IL-6. These data demonstrate the feasibility of developing specific IL-6R antagonists. The availability of such antagonists may offer an approach to specifically inhibit IL-6 activity in vivo.
- Subjects :
- Herpesvirus 4, Human
Carcinoma, Hepatocellular
Immunology
Molecular Sequence Data
Polymerase Chain Reaction
Mice
Mutant protein
Tumor Cells, Cultured
Immunology and Allergy
Animals
Humans
Amino Acid Sequence
Interleukin 6
Receptor
Cell Line, Transformed
DNA Primers
B-Lymphocytes
biology
Base Sequence
Interleukin-6
Liver Neoplasms
Oncostatin M
Interleukin
Receptors, Interleukin
Articles
Glycoprotein 130
Molecular biology
Receptors, Interleukin-6
Recombinant Proteins
Kinetics
Interleukin-6 receptor
biology.protein
Mutagenesis, Site-Directed
Multiple Myeloma
Leukemia inhibitory factor
Cell Division
Signal Transduction
Subjects
Details
- ISSN :
- 00221007
- Volume :
- 180
- Issue :
- 6
- Database :
- OpenAIRE
- Journal :
- The Journal of experimental medicine
- Accession number :
- edsair.doi.dedup.....418836c2177c0c78472114049c62c08a