Liu Hsu-Chung, Pai Shun-Yuan, Chen Hsiao-Ling, Lai Cheng-Wei, Tsai Tung-Chou, Cheng Winston T K, Yang Shang-Hsun, Chen Chuan-Mu
Department of Life Sciences, Agricultural Biotechnology Center, National Chung Hsing University, Taichung 402, Taiwan; Division of Chest Medicine, Department of Internal Medicine, Cheng Ching Hospital, Taichung 404, Taiwan; School of Medicine, Chung Shan Medical University, Taichung 404, Taiwan.
Department of Life Sciences, Agricultural Biotechnology Center, National Chung Hsing University, Taichung 402, Taiwan.
J Dairy Sci. 2014 Nov;97(11):6792-803. doi: 10.3168/jds.2014-8484. Epub 2014 Sep 6.
Recent advances in recombinant technology make transgenic animals that produce pharmaceutical proteins in their milk more feasible. The group 5 allergen isolated from Dermatophagoides pteronyssinus (Derp5) is one of the most important dust mite allergens in humans. The aims of this study were to develop transgenic mice that could secrete recombinant Derp5-containing milk and to demonstrate that ingesting recombinant milk protects against allergic airway inflammation. Two transgenes were constructed separately. The α-LA-Derp5f transgene consisted of the bovine α-lactalbumin (α-LA) promoter and full-length Derp5 cDNA. The α-LA-CN-Derp5t transgene included the α-LA promoter, a leader sequence of αS1-casein (CN), and signal peptide-truncated Derp5 cDNA. Both species of transgenic mice were confirmed to have successful transgene integration and stable germline transmission. Western blot analysis of the milk obtained from the offspring of transgenic mice demonstrated that recombinant Derp5 was secreted successfully in the milk of αLA-CN-Derp5t transgenic mice but not in that of αLA-Derp5f transgenic mice. This study provides new evidence that transgenic mice can secrete recombinant Derp5 efficiently in milk by adding a signal peptide of αS1-casein. The antigenic activity of recombinant Derp5 milk was demonstrated to have a protective effect against allergic airway inflammation in a murine model in which the ingestion of recombinant Derp5-containing milk was used as pretreatment.
重组技术的最新进展使能在乳汁中产生药用蛋白的转基因动物变得更加可行。从尘螨（Derp5）中分离出的第5组变应原是人类最重要的尘螨变应原之一。本研究的目的是培育能分泌含重组Derp5乳汁的转基因小鼠，并证明摄入重组乳汁可预防过敏性气道炎症。分别构建了两个转基因。α-LA-Derp5f转基因由牛α-乳白蛋白（α-LA）启动子和全长Derp5 cDNA组成。α-LA-CN-Derp5t转基因包括α-LA启动子、αS1-酪蛋白（CN）的前导序列和信号肽截短的Derp5 cDNA。两种转基因小鼠均被证实成功整合了转基因并实现了稳定的种系传递。对转基因小鼠后代的乳汁进行蛋白质印迹分析表明，重组Derp5在αLA-CN-Derp5t转基因小鼠的乳汁中成功分泌，但在αLA-Derp5f转基因小鼠的乳汁中未成功分泌。本研究提供了新的证据，即通过添加αS1-酪蛋白的信号肽，转基因小鼠可在乳汁中高效分泌重组Derp5。在以摄入含重组Derp5的乳汁作为预处理的小鼠模型中，重组Derp5乳汁的抗原活性被证明对过敏性气道炎症具有保护作用。
