Production of influenza vaccine using genetically manipulated chickens

使用基因改造鸡生产流感疫苗

• 批准号: 23656523
• 负责人: NISHIJIMA Kenichi
• 金额: $ 2.5万
• 依托单位: Nagoya University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Challenging Exploratory Research
• 财政年份: 2011
• 资助国家: 日本
• 起止时间: 2011 至 2012
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-23656523/
• 关键词: 生物機能工学; 感染症; 糖鎖; トランスジェニックニワトリ; ワクチン; 鶏卵

Vaccine against influenza virus is currently produced in developing chicken eggs. Allantoic membrane, the site for virus propagation for vaccine, does not express α2,6-linked sialic acid on cell surface, resulting poor infection/growth of human influenza virus. Therefore, chicken eggs with α2,6-linked sialic acid on allantoic membrane might be useful tool for vaccine production. To genetically modify chickens to express α2,6-linked sialic acid, the sialyltransferase gene was cloned and its enzymatic activity was confirmed. We also confirmed that ubiquitous actin promoter effectively expressed eGFP and single-chianed antibody in allantoic membrane. The sialyltransferase gene was inserted into the downstream of actin promoter of retrovirus vector construct. The packaging cell lines that produce high titer of retrovirus vector were successfully established. These are helpful to make transgenic chickens that express modified glycans on allantoic membrane.

流感病毒疫苗目前是在研制鸡蛋中生产的。尿囊膜是疫苗的病毒繁殖位点，在细胞表面不表达α2,6-链唾液酸，导致人流感病毒感染/生长不良。因此，在尿囊膜上添加α2,6-链唾液酸的鸡蛋可能是疫苗生产的有效工具。为了改良鸡表达α2,6-链唾液酸，克隆了唾液基转移酶基因，并对其酶活性进行了验证。我们还证实了无处不在的肌动蛋白启动子在尿囊膜上有效表达eGFP和单链抗体。将唾液基转移酶基因插入逆转录病毒载体的肌动蛋白启动子下游。成功建立了生产高滴度逆转录病毒载体的包装细胞系。这有助于制备在尿囊膜上表达修饰聚糖的转基因鸡。

项目成果

Transgenic chickens, Development

转基因鸡，发育

• 发表时间: 2013
• 期刊: Growth & Differentiation
• 作者: J. Zhang;S. Yamaguchi;S. Inoue;H.Hirakawa;T. Nagamune;Ken-ichi Nishijima and Shinji Iijima
• 通讯作者: Ken-ichi Nishijima and Shinji Iijima

ニワトリ卵白に生産させた医薬用タンパク質の糖鎖制御

鸡蛋清生产的药用蛋白的糖链控制

• 发表时间: 2011
• 期刊: バイオサイエンスとインダストリー
• 作者: X. Liu;M. Eichenberger;Y. Fujioka;J. Dong;and H. Ueda;西島謙一
• 通讯作者: 西島謙一