Kaori Miyazaki-Komine1,2, Yoshiki Takai3, Ping Huang1, Osamu Kusano-Arai4,5, Hiroko Iwanari4, Tatsuro Misu6, Katsushi Koda7, Katsuyuki Mitomo7, Toshiko Sakihama4, Yoshiaki Toyama2, Kazuo Fujihara6, Takao Hamakubo4, Masato Yasui1,8, Yoichiro Abe1,8. 1. Department of Pharmacology, School of Medicine, Keio University, 35 Shinanomachi, Shinjuku-ku, Tokyo, 160-8582, Japan. 2. Department of Orthopaedic Surgery, School of Medicine, Keio University, Tokyo, Japan. 3. Department of Neurology, Tohoku University School of Medicine, 1-1 Seiryomachi, Aoba-ku, Sendai, 980-8574, Japan. 4. Quantitative Biology and Medicine, Research Center for Advanced Science and Technology, The University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo, 153-8904, Japan. 5. Institute of Immunology Co., Ltd., 1-1-10 Koraku, Bunkyo-ku, Tokyo, 112-0004, Japan. 6. Department of Multiple Sclerosis Therapeutics, Tohoku University Graduate School of Medicine, 1-1 Seiryomachi, Aoba-ku, Sendai, 980-8574, Japan. 7. Research and Development Division, Perseus Proteomics Inc., 4-7-6 Komaba, Meguro-ku, Tokyo, 153-0041, Japan. 8. Keio Advanced Research Center for Water Biology and Medicine, Keio University, Tokyo, Japan.
Abstract
BACKGROUND AND PURPOSE: Most of the cases of neuromyelitis optica (NMO) are characterized by the presence of an autoantibody, NMO-IgG, which recognizes the extracellular domains of the water channel, aquaporin-4. Binding of NMO-IgG to aquaporin-4 expressed in end-feet of astrocytes leads to complement-dependent disruption of astrocytes followed by demyelination. One therapeutic option for NMO is to prevent the binding of NMO-IgG to aquaporin-4, using high-avidity, non-pathogenic-chimeric, monoclonal antibodies to this water channel. We describe here the development of such antibodies. EXPERIMENTAL APPROACH: cDNAs encoding variable regions of heavy and light chains of monoclonal antibodies against the extracellular domains of human aquaporin-4 were cloned from hybridoma total RNA and fused to those encoding constant regions of human IgG1 and Igκ respectively. Then mammalian expression vectors were constructed to establish stable cell lines secreting mature chimeric antibodies. KEY RESULTS: Original monoclonal antibodies showed high avidity binding to human aquaporin-4, as determined by ELISA. Live imaging using Alexa-Fluor-555-labelled antibodies revealed that the antibody D15107 more rapidly bound to cells expressing human aquaporin-4 than others and strongly enhanced endocytosis of this water channel, while D12092 also bound rapidly to human aquaporin-4 but enhanced endocytosis to a lesser degree. Chimeric D15107 prevented complement-dependent cytotoxicity induced by NMO-IgG from patient sera in vitro. CONCLUSIONS AND IMPLICATIONS: We have established non-pathogenic, high-avidity, chimeric antibodies against the extracellular domains of human aquaporin-4, which provide a novel therapeutic option for preventing the progress and recurrence of NMO/NMO spectrum disorders.
BACKGROUND AND PURPOSE: Most of the cases of neuromyelitis optica (NMO) are characterized by the presence of an autoantibody, NMO-IgG, which recognizes the extracellular domains of the water channel, aquaporin-4. Binding of NMO-IgG to aquaporin-4 expressed in end-feet of astrocytes leads to complement-dependent disruption of astrocytes followed by demyelination. One therapeutic option for NMO is to prevent the binding of NMO-IgG to aquaporin-4, using high-avidity, non-pathogenic-chimeric, monoclonal antibodies to this water channel. We describe here the development of such antibodies. EXPERIMENTAL APPROACH: cDNAs encoding variable regions of heavy and light chains of monoclonal antibodies against the extracellular domains of humanaquaporin-4 were cloned from hybridoma total RNA and fused to those encoding constant regions of human IgG1 and Igκ respectively. Then mammalianexpression vectors were constructed to establish stable cell lines secreting mature chimeric antibodies. KEY RESULTS: Original monoclonal antibodies showed high avidity binding to humanaquaporin-4, as determined by ELISA. Live imaging using Alexa-Fluor-555-labelled antibodies revealed that the antibody D15107 more rapidly bound to cells expressing humanaquaporin-4 than others and strongly enhanced endocytosis of this water channel, while D12092 also bound rapidly to humanaquaporin-4 but enhanced endocytosis to a lesser degree. Chimeric D15107 prevented complement-dependent cytotoxicity induced by NMO-IgG from patient sera in vitro. CONCLUSIONS AND IMPLICATIONS: We have established non-pathogenic, high-avidity, chimeric antibodies against the extracellular domains of humanaquaporin-4, which provide a novel therapeutic option for preventing the progress and recurrence of NMO/NMO spectrum disorders.
Authors: Vanda A Lennon; Dean M Wingerchuk; Thomas J Kryzer; Sean J Pittock; Claudia F Lucchinetti; Kazuo Fujihara; Ichiro Nakashima; Brian G Weinshenker Journal: Lancet Date: 2004 Dec 11-17 Impact factor: 79.321
Authors: Samira Saadoun; Patrick Waters; B Anthony Bell; Angela Vincent; A S Verkman; Marios C Papadopoulos Journal: Brain Date: 2010-01-04 Impact factor: 13.501