重组HIV-1 gp120 AAV2/1在小鼠和恒河猴体内的免疫原性  被引量:1

Immunogenicity of Recombinant Adeno-asscociated Virus Type 1 Expressing HIV-1 gp120 Gene in Mice and Rhesus Macaques

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作  者:余双庆[1] 冯霞[1] 刘红梅[1] 杨海儒[1] 李红霞[1] 曾毅[1] 

机构地区:[1]中国疾病预防控制中心病毒病预防控制所传染病预防控制国家重点实验室,北京100052

出  处:《病毒学报》2010年第2期115-120,共6页Chinese Journal of Virology

基  金:国家重大科技专项(2008ZX10001-009);国家自然科学基金(30872397)

摘  要:本文旨在研究表达HIV-1中国流行株gp120基因的重组腺相关病毒疫苗在小鼠和恒河猴体内的免疫原性。用rAAV2/1-gp120免疫BALB/c小鼠和恒河猴,分别用ELISA和HIV-1假病毒中和试验检测免疫动物血清中HIV-1特异性IgG抗体水平和中和抗体水平,用ELISPOT方法和体内杀伤实验检测HIV-1特异性细胞免疫应答水平。结果显示在小鼠体内用rAAV2/1-gp120仅免疫一次就能诱导较高水平的IgG抗体,IgG抗体至少能持续21周,但没有检测到中和抗体。rAAV2/1-gp120在小鼠体内诱导微弱水平的HIV-1特异性细胞免疫应答。rAAV2/1-gp120在恒河猴体内诱导的HIV-1特异性细胞和抗体反应均很弱,且检测不到中和抗体。提示rAAV2/1载体在诱导抗体反应方面具有特殊优势,但若希望诱导HIV-1特异性中和抗体,则需要改造env基因以提高其免疫原性。To study the immunogenicity of recombinant adeno-asscociated virus type 1 expressing HIV-1 gp120 gene (rAAV2/1-gp120) in BALB/c mice and Rhesus macaques. The gp120 gene derived from Chinese HIV-1 isolates was constructed into rAAV2/1 and rAd5 vectors. Firstly,the immunogenicity of rAAV2/1-gp120 was compared with rAd5-gp120 in BALB/c mice when used once or twice in 3 weeks interval. Then the monkeys were immunized with rAAV2/1-gp120 once. The HIV-1 specific IgG levels and neutralization activity to pseudotyped HIV-1 virus were tested using ELISA and neutralization assay,and the cellular immune responses were analyzed by IFN-γ enzyme-linked immunospot (ELISPOT) and in vivo CTL assays. Compared with rAd5-gp120 immunized mice,mice immunized with rAAV2/1-gp120 once induced stronger gp120-specific IgG and were sustained for at least 21 weeks. rAd5-gp120 immunized mice generated stronger cellular immune responses than rAAV2/1-gp120 in spleen and draining lymph node. But only moderate gp120-specific in vivo CTL activity was observed in both rAAV2/1-gp120 and rAd5-gp120 immunized mice. Four of five monkeys vaccinated with rAAV2/1-gp120 generated gp120 specific IgG,the titer ranged from 1:100 to 1:400 with end-point dilution. Gp120 specific IgG could be detected 4 weeks after immunization and reached the peak at 10 weeks after immunization. No neutralization activity against pseudotyped HIV-1virus expressing NL4-3 Env antigen was detected. In Conclusion,rAAV2/1-gp120 induced high level of HIV-1 specific IgG antibody and moderate cellular immune responses. No neutralizing antibody was elicited. It indicates that the env gene and immunization strategy should be optimized to elicit neutralizing antibody against HIV-1 in further studies.

关 键 词:HIV-1 包膜糖蛋白Gp120 重组腺相关病毒 免疫原性 

分 类 号:R373.9[医药卫生—病原生物学] Q78[医药卫生—基础医学]

 

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