title: A rice-based soluble form of a murine TNF-specific llama variable domain of heavy-chain antibody suppresses collagen-induced arthritis in mice.
abstract: Tumor necrosis factor alpha (TNF) plays a pivotal role in chronic inflammatory diseases such as rheumatoid arthritis and Crohn's disease. Although anti-TNF antibody therapy is now commonly used to treat patients suffering from these inflammatory conditions, the cost of treatment continues to be a concern. Here, we developed a rice transgenic system for the production of a llama variable domain of a heavy-chain antibody fragment (VHH) specific for mouse TNF in rice seeds (MucoRice-mTNF-VHH). MucoRice-mTNF-VHH was produced at high levels in the rice seeds when we used our most recent transgene-overexpression system with RNA interference technology that suppresses the production of major rice endogenous storage proteins while enhancing the expression of the transgene-derived protein. Production levels of mTNF-VHH in rice seeds reached an average of 1.45% (w/w). Further, approximately 91% of mTNF-VHH was released easily when the powder form of MucoRice-mTNF-VHH was mixed with PBS. mTNF-VHH purified by means of single-step gel filtration from rice PBS extract showed high neutralizing activity in an in vitro mTNF cytotoxicity assay using WEHI164 cells. In addition, purified mTNF-VHH suppressed progression of collagen-induced arthritis in mice. These results show that this rice-expression system is useful for the production of neutralizing VHH antibody specific for mTNF.
authors.affiliation: Division of Mucosal Immunology, The Institute of Medical Science, The University of Tokyo, 4-6-1, Shirokanedai, Minato-ku, Tokyo, Japan; MG Pharma Inc., 7-7-25, Saito-Asagi, Ibaraki, Osaka, Japan.
authors.forename: Michiyo
authors.initials: M
authors.lastname: Abe
authors.affiliation: Division of Mucosal Immunology, The Institute of Medical Science, The University of Tokyo, 4-6-1, Shirokanedai, Minato-ku, Tokyo, Japan; International Research and Development Center for Mucosal Vaccine, The Institute of Medical Science, The University of Tokyo, 4-6-1, Shirokanedai, Minato-ku, Tokyo, Japan. Electronic address: yukiy@ims.u-tokyo.ac.jp.
authors.forename: Yoshikazu
authors.initials: Y
authors.lastname: Yuki
authors.affiliation: Division of Mucosal Immunology, The Institute of Medical Science, The University of Tokyo, 4-6-1, Shirokanedai, Minato-ku, Tokyo, Japan.
authors.forename: Shiho
authors.initials: S
authors.lastname: Kurokawa
authors.affiliation: Division of Mucosal Immunology, The Institute of Medical Science, The University of Tokyo, 4-6-1, Shirokanedai, Minato-ku, Tokyo, Japan.
authors.forename: Mio
authors.initials: M
authors.lastname: Mejima
authors.affiliation: Crop Development Division, NARO Agriculture Research Center, 1-2-1, Inada, Joetsu, Niigata, Japan.
authors.forename: Masaharu
authors.initials: M
authors.lastname: Kuroda
authors.affiliation: Division of Mucosal Immunology, The Institute of Medical Science, The University of Tokyo, 4-6-1, Shirokanedai, Minato-ku, Tokyo, Japan.
authors.forename: Eun Jeong
authors.initials: EJ
authors.lastname: Park
authors.affiliation: Institute for Biochemistry and Molecular Biology II, Medical Faculty, Heinrich-Heine-University Düsseldorf, Düsseldorf 40225, Germany.
authors.forename: Jürgen
authors.initials: J
authors.lastname: Scheller
authors.affiliation: MG Pharma Inc., 7-7-25, Saito-Asagi, Ibaraki, Osaka, Japan.
authors.forename: Ushio
authors.initials: U
authors.lastname: Nakanishi
authors.affiliation: Division of Mucosal Immunology, The Institute of Medical Science, The University of Tokyo, 4-6-1, Shirokanedai, Minato-ku, Tokyo, Japan; International Research and Development Center for Mucosal Vaccine, The Institute of Medical Science, The University of Tokyo, 4-6-1, Shirokanedai, Minato-ku, Tokyo, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Kiyono
mesh_terms: D000818:Animals; D018501:Antirheumatic Agents / Q000008:administration & dosage* / Q000627:therapeutic use; D001169:Arthritis, Experimental / Q000276:immunology / Q000628:therapy*; D002161:Camelids, New World / Q000276:immunology; D007143:Immunoglobulin Heavy Chains / Q000276:immunology*; D007135:Immunoglobulin Variable Region / Q000276:immunology*; D008297:Male; D051379:Mice; D012275:Oryza / Q000235:genetics* / Q000276:immunology; D030821:Plants, Genetically Modified; D012639:Seeds / Q000235:genetics / Q000276:immunology; D012995:Solubility; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000276:immunology*
chemical_list: D018501:Antirheumatic Agents; D007143:Immunoglobulin Heavy Chains; D007135:Immunoglobulin Variable Region; D014409:Tumor Necrosis Factor-alpha
keywords: MucoRice; Nanobody; Plant; Single domain antibody; Tumor necrosis factor