Production of influenza vaccine using genetically manipulated chickens

目前，针对流感病毒的疫苗正在开发中的鸡蛋中生产。疫苗的病毒繁殖部位尿囊膜在细胞表面不表达α-2，6-联结唾液酸，导致人流感病毒感染/生长不良。因此，尿囊膜上含有α-2，6-联结唾液酸的鸡蛋可能是疫苗生产的有用工具。为了对鸡进行α-2，6-连接唾液酸的基因改造，克隆了唾液酸基转移酶基因，并对其活性进行了鉴定。我们还证实了普遍存在的肌动蛋白启动子在尿囊膜中有效地表达了EGFP和单链抗体。将唾液酸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。这为制备在尿囊膜上表达修饰多糖的转基因鸡奠定了基础。

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.