Tian Guobin, Zhang Suhua, Li Yanbing, Bu Zhigao, Liu Peihong, Zhou Jinping, Li Chengjun, Shi Jianzhong, Yu Kangzhen, Chen Hualan
Animal Influenza Laboratory of the Ministry of Agriculture, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, PR China.
Virology. 2005 Oct 10;341(1):153-62. doi: 10.1016/j.virol.2005.07.011.
We generated a high-growth H5N1/PR8 virus by plasmid-based reverse genetics. The virulence associated multiple basic amino acids of the HA gene were removed, and the resulting virus is attenuated for chickens and chicken eggs. A formalin-inactivated oil-emulsion vaccine was prepared from this virus. When SPF chickens were inoculated with 0.3 ml of the vaccine, the hemagglutinin-inhibition (HI) antibody became detectable at 1 week post-vaccination (p.v.) and reached a peak of 10log2 at 6 weeks p.v. then slowly declined to 4log2 at 43 weeks p.v. Challenge studies performed at 2, 3 and 43 weeks p.v. indicated that all of the chickens were completely protected from disease signs and death. Ducks and geese were completely protected from highly pathogenic H5N1 virus challenge 3 weeks p.v. The duration of protective immunity in ducks and geese was investigated by detecting the HI antibody of the field vaccinated birds, and the results indicated that 3 doses of the vaccine inoculation in geese could induce a 34 weeks protection, while 2 doses induced more than 52 weeks protection in ducks. We first reported that an oil-emulsion inactivated vaccine derived from a high-growth H5N1 vaccine induced approximately 10 months of protective immunity in chickens and demonstrated that the oil-emulsion inactivated avian influenza vaccine is immunogenic for geese and ducks. These results provide useful information for the application of vaccines to the control of H5N1 avian influenza in poultry, including chickens and domestic waterfowl.
我们通过基于质粒的反向遗传学技术构建了一种高生长性的H5N1/PR8病毒。去除了HA基因中与毒力相关的多个碱性氨基酸，所得病毒对鸡和鸡蛋的致病性减弱。用这种病毒制备了福尔马林灭活油乳剂疫苗。当给SPF鸡接种0.3 ml该疫苗时，接种后1周可检测到血凝抑制（HI）抗体，接种后6周达到峰值10log2，然后在接种后43周缓慢降至4log2。在接种后2、3和43周进行的攻毒试验表明，所有鸡均完全免受疾病症状和死亡的影响。接种后3周，鸭和鹅完全免受高致病性H5N1病毒攻击。通过检测现场接种疫苗禽类的HI抗体，研究了鸭和鹅的保护性免疫持续时间，结果表明，鹅接种3剂疫苗可诱导34周的保护，而鸭接种2剂疫苗可诱导超过52周的保护。我们首次报道，一种源自高生长性H5N1疫苗的油乳剂灭活疫苗可在鸡中诱导约10个月的保护性免疫，并证明该油乳剂灭活禽流感疫苗对鹅和鸭具有免疫原性。这些结果为疫苗在控制包括鸡和家鸭在内的家禽H5N1禽流感中的应用提供了有用信息。
