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Secretion of an immunoreactive single-chain variable fragment antibody against mouse interleukin 6 by Lactococcus lactis

  • Applied microbial and cell physiology
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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.

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Acknowledgments

We are grateful to Yasuhiro Yasaki, Kohichi Sudo, and Fu Namai (Faculty of Agriculture, Shinshu University) for their excellent support in purification of recombinant proteins.

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Authors and Affiliations

  1. Department of Bioscience and Food Production Science, Interdisciplinary Graduate School of Science and Technology, Shinshu University, 8304 Minamiminowa, Kamiina, Nagano, 399-4598, Japan

    Suguru Shigemori, Masaki Ihara, Yoshinari Yamamoto, Shireen Nigar & Takeshi Shimosato

  2. 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

    Suguru Shigemori & Yoshinari Yamamoto

  3. Department of Interdisciplinary Genome Sciences and Cell Metabolism, Institute for Biomedical Sciences, Shinshu University, 8304 Minamiminowa, Kamiina, Nagano, 399-4598, Japan

    Masaki Ihara, Tasuku Ogita & Takeshi Shimosato

  4. Department of Internal Medicine and Clinical Immunology, Graduate School of Medicine, Yokohama City University, 3-9 Fukuura, Kanazawa-ku, Yokohama, Kanagawa, 236-0004, Japan

    Takashi Sato

  5. Department of Nutrition and Food Technology, Jessore University of Science and Technology, Churamonkathi-Chowgacha Rd, Jessore, 7408, Bangladesh

    Shireen Nigar

  6. Supramolecular Complexes Unit, Research Center for Fungal and Microbial Dynamism, Shinshu University, 8304 Minamiminowa, Kamiina, Nagano, 399-4598, Japan

    Takeshi Shimosato

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  1. Suguru Shigemori
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  2. Masaki Ihara
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  3. Takashi Sato
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  4. Yoshinari Yamamoto
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  5. Shireen Nigar
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  6. Tasuku Ogita
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  7. Takeshi Shimosato
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Correspondence to Takeshi Shimosato.

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Funding

This study was supported by a Grant-in-Aid for the Japan Society for the Promotion of Science Fellows (No. 14J06317) to SS and by a grant from Sumitomo Electric Industries Group CSR Foundation (No. 2012#7) to TSh.

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The authors declare that they have no conflict of interest.

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This article does not contain any studies with human participants or animals performed by any of the authors.

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Shigemori, S., Ihara, M., Sato, T. et al. Secretion of an immunoreactive single-chain variable fragment antibody against mouse interleukin 6 by Lactococcus lactis . Appl Microbiol Biotechnol 101, 341–349 (2017). https://doi.org/10.1007/s00253-016-7907-8

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