Secretion of an immunoreactive single-chain variable fragment antibody against mouse interleukin 6 by Lactococcus lactis
Secretion of an immunoreactive single-chain variable fragment antibody against mouse interleukin 6 by Lactococcus lactis
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DOI：
10.1007/s00253-016-7907-8
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发表时间：
2017-01-01
影响因子：
5
通讯作者：
Shimosato, Takeshi
中科院分区：
文献类型：
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作者：
Shigemori, Suguru;Ihara, Masaki;Shimosato, Takeshi
白细胞介素6（IL-6）是多种炎症性疾病、自身免疫性疾病和癌症的重要致病因子。针对与IL-6/IL-6受体信号传导相关的分子的阻断抗体是预防或治疗这些疾病的有吸引力的候选物。在这项研究中，我们开发了一种转基因乳酸乳球菌分泌的单链抗体抗小鼠IL-6（IL-6scFv）。通过将IL 6scFv基因片段克隆到乳球菌分泌质粒中构建IL 6scFv分泌载体，并电穿孔到L.乳酸菌NZ 9000（NZ-IL 6scFv）。免疫印迹法证实nisin诱导的NZ-IL 6scFv分泌重组IL 6scFv（rIL 6scFv），并通过调节培养条件进行优化。我们发现rIL-6scFv可以与商品化的重组小鼠IL-6结合。该结果清楚地证明了rIL 6scFv的免疫反应性。这是第一项工程化乳酸菌（gmLAB）的转基因菌株的研究，该菌株产生功能性抗细胞因子scFv。许多先前的研究表明，使用gmLAB粘膜递送生物医学蛋白是治疗各种疾病的有效且低成本的方法。因此，NZ-IL 6scFv可能是研究和开发用于各种炎症和自身免疫性疾病以及癌症的新IL-6靶向剂的有吸引力的工具。
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.