H5-H7双价禽流感核酸疫苗免疫研究  

Study on construction and experimental immunity of H5,H7 subtype influenza virus bivalent DNA vaccine

作　　者：张丹[1,2] 鲁会军[2] 谭磊[1,2] 刘存霞[1,2] 胡博[1,2] 刘燕瑜[1,2] 刘昊[1,2] 金扩世[2] 金宁一[2] 

机构地区：[1]吉林大学畜牧兽医学院,吉林长春130062 [2]解放军军事医学科学院军事兽医研究所全军基因工程重点实验室,吉林长春130062

出　　处：《中国兽医学报》2011年第2期184-188,共5页Chinese Journal of Veterinary Science

基　　金："863"重大项目(2006AA10A205);国家科技支撑项目(2006BAD06A05)

摘　　要：由于禽流感的高度变异性,免疫后不同亚型之间难以得到交叉保护,以致病毒得以逃避宿主免疫系统的监视,多价核酸禽流感疫苗具有可以保护不同亚型病毒攻击的特点。本试验设计并构建了包含禽流感H5HA和H7HA1基因的双价真核表达质粒pV-H5-H7及单独表达H5HA和H7HA1的pV-H5和pV-H7HA1。通过RT-PCR,间接免疫荧光（IFA）等方法验证构建质粒的正确性和其表达蛋白的免疫原性。0,21 d分别免疫6~8周龄的BALB/c小鼠,设立双价疫苗组,单表达免疫组和对照组。免疫后35 d用HPAIV H5N1进行致死性攻击。结果显示免疫组均可刺激机体产生H5特异性抗体,pV-H5-H7诱导产生的抗体对H5N1的攻毒保护率为80%,而pV-H5单表达的攻毒保护率也为80%。表明本实验构建的双价禽流感核酸疫苗的免疫效果与单表达组相当（P〉0.05）,为多价禽流感核酸疫苗的研制提供了基础。It is difficult to provide cross-protection among different subtypes after immunization because of avian influenza virus with high variability,which cause the virus could escape the host immune surveillance.Multivalence DNA vaccine of avian influenza may protect animals from different subtypes.This experimental design was constructed to contain the avian influenza virus H5HA and H7HA1 bivalent-gene plasmid pV-H5-H7 and single-gene H7HA1,H5HA named pV-H5 and pV-H7HA1.By RT-PCR and indirect immunofluorescence（IFA）,the correctness of constructed plasmids and their proteins immunogenicity were validated.6-8 weeks old BALB/c mice were immunized on 0 day and 21 day,establishment of bivalent vaccine group（pV-H5-H7）,single-gene immune groups（pV-H5,pV-H7HA1） and the control group（pVAX1）.35 days post innoculation,challenge with lethal dose HPAIV H5N1 virus.The results showed that immunized groups can stimulate the mice to produce specific IgG antibodies against H5 subtype.The survival rate of pV-H5-H7 group against H5N1 virus challenge was 80%,equal to single expression of pV-H5 group.This study showed that the H5-H7 bivalent vaccine group and single-gene group against avian influenza virus was not significant（P0.05）,which provide the foundation on multivalent DNA vaccine against avian influenza virus.

关 键 词：禽流感 H5 H7亚型 核酸疫苗 免疫攻毒 

分 类 号：S852.65[农业科学—基础兽医学]