摘要
目的观察乙型肝炎病毒(HBV)外膜大蛋白(LP)基因的2型重组腺相关病毒(rAAV-2-LP)在Balb/c小鼠体内的免疫原性。方法以5×1011的rAAV-2-LP单次肌肉注射接种Balb/c小鼠,RT-PCR扩增肝脏组织中HBV外膜大蛋白基因cDNA;RIA法检测血清中表面抗体(抗-HBs)滴度;51Cr释放分析检测细胞毒性T淋巴细胞(CTL)活性。结果HBV外膜大蛋白基因已整合到肝细胞染色体DNA,rAAV-LP接种小鼠后第20 d,小鼠体内可出现针对HBV外膜大蛋白特异性CTL,第30 d,血清中可检测到表面抗体。结论rAAV可以介导HBV外膜大蛋白基因转移到小鼠肝细胞;rAAV-2-LP免疫小鼠可以同时诱导体液和细胞免疫反应的出现。基于rAAV-2载体的HBV疫苗,对于防止HBV感染,尤其是作为慢性乙型肝炎的治疗性疫苗可能具有潜在的应用价值。
Objective To investigate the immunogenicity of recombinant adenoI associated virus type 2 （rAAV-2） expressing Hepatitis B virus envelope large protein （rAAV-2-LP） in vivo. Methods One single dose of rAAV-2-LP（5 ×10^11 virus particles） was inculoated to Balb/c mice via intramuscular injection. At 30th day, cDNA fragment of large protein in liver tissue was amplified by RT-PCR. The HBsAg antibody（Anti-HBs） in sera was assayed by radioimmunoassay（RIA）. Cytotoxic Tlymphocyte（CTL） assays were performed with 51 Cr release assay. Results Exogenous large protein gene can be integrated into the hepatocellular chromosomal DNA. At 20^th day after inoculation with rAAV-LP, specific CTL activity can be detected in mice. At 3Dth day, Anti-HBs in sera can be detected, Conclusion rAAV mediated exogenous HBV envelope large protein gene can be transferred into cells in liver tissue, rAAV-2-LP can induced both humoral and cellular immune response. The results indicated that the rAAV-2-LP can be used as a promising candidate for hepatitis B vaccine, particularly as a therapeutic vaccine for chronic hepatitis B.
出处
《中国公共卫生》
CAS
CSCD
北大核心
2006年第11期1357-1358,共2页
Chinese Journal of Public Health
基金
广东省医学科研基金(A2004366)
