鸭乙型肝炎病毒核心抗原DNA 疫苗的构建及其诱导的体液免疫应答

蒋定锋; 闫梁; 贲海静; 赵艳丰; 秦智强; 瞿涤

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微生物与感染 ›› 2009, Vol. 4 ›› Issue (3) : 140-145.

论著

鸭乙型肝炎病毒核心抗原DNA 疫苗的构建及其诱导的体液免疫应答

蒋定锋；闫梁；贲海静；赵艳丰；秦智强；瞿涤

作者信息 +

Construction of duck hepatitis B virus core antigen DNA vaccine and
induced humoral immune response

JIANG Ding-Feng；YAN Liang；BEN Hai-Jing；ZHAO Yan-Feng；QIN Zhi-Qiang；QU Di

Author information +

文章历史 +

摘要

为研究鸭乙型肝炎病毒核心抗原( DHBcAg) 真核表达质粒的免疫原性, 分析DHBcAg DNA 疫苗诱导的体液免疫应答, 首先借助生物信息学方法对鸭乙型肝炎病毒( DHBV) Core 基因编码的氨基酸序列进行亲疏水性分析, 分别构建DHBcAg 全基因( E-DHBc263) 及去除疏水性序列的DHBcAg 片段( E-DHBc180) 的真核表达质粒, 间接免疫荧光检测结果显示可在COS7 细胞内表达。进一步构建原核表达质粒p-DHBc263 和p-DHBc180, 仅p-DHBc180 可表达蛋白, 纯化后作为酶联免疫吸附试验( ELISA) 包被抗原, 用于DHBcAb 的检测。分别用E-DHBc263 和E-DHBc180 免疫小鼠, 采用间接ELISA 检测DHBcAb。结果显示, E-DHBc180 可诱导免疫小鼠产生DHBcAb 免疫应答, 加强免疫后效价可达1∶100 ～1∶400。结果提示, E-DHBc180 可作为DHBcAg DNA 疫苗, 在DHBV 感染鸭模型中评价其效果。

Abstract

To investigate the immunogenicity of eukaryotic expression plasmid of duck hepatitis B virus core antigen ( DHBcAg) in mice, we first analyzed the hydrophilic and hydrophobic characteristics of the amino acid sequence of
DHBcAg by bioinformatics with the Vector NTI Suite software, and constructed the eukaryotic expression plasmids of the complete ( E-DHBc 263) and partial sequences ( only the hydrophilic part, E-DHBc 180) of DHBcAg gene. The
results showed that both plasmids expressed DHBcAg in transfected COS7 cells detected by indirect immunofluorescence. The prokaryotic expression plasmids, p-DHBc 263 and p-DHBc 180, were constructed to express DHBcAg for DHBcAb detection, but only p-DHBc180-expressed DHBcAg could be detected by Western blot. Then we immunized BALB/ c mice with E-DHBc 180 and E-DHBc 263. DHBcAb titers in sera from immunized mice were detected by enzyme-linked immunosorbent assay ( ELISA) on the plates coated with prokaryotically expressed DHBcAg. The mice primed and boosted with E-DHBc180 produced DHBcAb, and titers were 1∶100-1∶400, while in
E-DHBc 263-immunized mice DHBcAb level was belowthe detection level. It suggests that E-DHBc 180 could be used as DHBcAg DNA vaccine for further research in its effects on DHBV infection.

导出引用

蒋定锋；闫梁；贲海静；赵艳丰；秦智强；瞿涤. 鸭乙型肝炎病毒核心抗原DNA 疫苗的构建及其诱导的体液免疫应答[J]. 微生物与感染. 2009, 4(3): 140-145

JIANG Ding-Feng；YAN Liang；BEN Hai-Jing；ZHAO Yan-Feng；QIN Zhi-Qiang；QU Di. Construction of duck hepatitis B virus core antigen DNA vaccine and
induced humoral immune response[J]. Journal of Microbes and Infections. 2009, 4(3): 140-145