title: Secretion of an immunoreactive single-chain variable fragment antibody against mouse interleukin 6 by Lactococcus lactis.
abstract: Interleukin 6 (IL-6) is an important pathogenic factor in development of various inflammatory and autoimmune diseases and cancer. Blocking antibodies against molecules associated with IL-6/IL-6 receptor signaling are an attractive candidate for the prevention or therapy of these diseases. In this study, we developed a genetically modified strain of Lactococcus lactis secreting a single-chain variable fragment antibody against mouse IL-6 (IL6scFv). An IL6scFv-secretion vector was constructed by cloning an IL6scFv gene fragment into a lactococcal secretion plasmid and was electroporated into L. lactis NZ9000 (NZ-IL6scFv). Secretion of recombinant IL6scFv (rIL6scFv) by nisin-induced NZ-IL6scFv was confirmed by western blotting and was optimized by tuning culture conditions. We found that rIL6scFv could bind to commercial recombinant mouse IL-6. This result clearly demonstrated the immunoreactivity of rIL6scFv. This is the first study to engineer a genetically modified strain of lactic acid bacteria (gmLAB) that produces a functional anti-cytokine scFv. Numerous previous studies suggested that mucosal delivery of biomedical proteins using gmLAB is an effective and low-cost way to treat various disorders. Therefore, NZ-IL6scFv may be an attractive tool for the research and development of new IL-6 targeting agents for various inflammatory and autoimmune diseases as well as for cancer.
authors.affiliation: Department of Bioscience and Food Production Science, Interdisciplinary Graduate School of Science and Technology, Shinshu University, 8304 Minamiminowa, Kamiina, Nagano, 399-4598, Japan.|Research Fellow of the Japan Society for the Promotion of Science, Japan Society for the Promotion of Science, 5-3-1, Kojimachi, Chiyoda-ku, Tokyo, 102-0083, Japan.
authors.forename: Suguru
authors.initials: S
authors.lastname: Shigemori
authors.affiliation: Department of Bioscience and Food Production Science, Interdisciplinary Graduate School of Science and Technology, Shinshu University, 8304 Minamiminowa, Kamiina, Nagano, 399-4598, Japan.|Department of Interdisciplinary Genome Sciences and Cell Metabolism, Institute for Biomedical Sciences, Shinshu University, 8304 Minamiminowa, Kamiina, Nagano, 399-4598, Japan.
authors.forename: Masaki
authors.initials: M
authors.lastname: Ihara
authors.affiliation: Department of Internal Medicine and Clinical Immunology, Graduate School of Medicine, Yokohama City University, 3-9 Fukuura, Kanazawa-ku, Yokohama, Kanagawa, 236-0004, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Sato
authors.affiliation: Department of Bioscience and Food Production Science, Interdisciplinary Graduate School of Science and Technology, Shinshu University, 8304 Minamiminowa, Kamiina, Nagano, 399-4598, Japan.|Research Fellow of the Japan Society for the Promotion of Science, Japan Society for the Promotion of Science, 5-3-1, Kojimachi, Chiyoda-ku, Tokyo, 102-0083, Japan.
authors.forename: Yoshinari
authors.initials: Y
authors.lastname: Yamamoto
authors.affiliation: Department of Bioscience and Food Production Science, Interdisciplinary Graduate School of Science and Technology, Shinshu University, 8304 Minamiminowa, Kamiina, Nagano, 399-4598, Japan.|Department of Nutrition and Food Technology, Jessore University of Science and Technology, Churamonkathi-Chowgacha Rd, Jessore, 7408, Bangladesh.
authors.forename: Shireen
authors.initials: S
authors.lastname: Nigar
authors.affiliation: Department of Interdisciplinary Genome Sciences and Cell Metabolism, Institute for Biomedical Sciences, Shinshu University, 8304 Minamiminowa, Kamiina, Nagano, 399-4598, Japan.
authors.forename: Tasuku
authors.initials: T
authors.lastname: Ogita
authors.affiliation: Department of Bioscience and Food Production Science, Interdisciplinary Graduate School of Science and Technology, Shinshu University, 8304 Minamiminowa, Kamiina, Nagano, 399-4598, Japan. shimot@shinshu-u.ac.jp.|Department of Interdisciplinary Genome Sciences and Cell Metabolism, Institute for Biomedical Sciences, Shinshu University, 8304 Minamiminowa, Kamiina, Nagano, 399-4598, Japan. shimot@shinshu-u.ac.jp.|Supramolecular Complexes Unit, Research Center for Fungal and Microbial Dynamism, Shinshu University, 8304 Minamiminowa, Kamiina, Nagano, 399-4598, Japan. shimot@shinshu-u.ac.jp.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Shimosato
mesh_terms: D000818:Animals; D003001:Cloning, Molecular; D018274:Electroporation; D015870:Gene Expression; D013294:Lactococcus lactis / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D010957:Plasmids; D011485:Protein Binding; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism*; D057127:Single-Chain Antibodies / Q000235:genetics / Q000378:metabolism*; D000093242:Interleukin-6 Inhibitors
chemical_list: D011994:Recombinant Proteins; D057127:Single-Chain Antibodies; D000093242:Interleukin-6 Inhibitors
keywords: Immunoreactivity; Interleukin 6; Lactococcus lactis; gmLAB; scFv antibody