HBV外膜蛋白2型重组腺相关病毒免疫原性

郝晓萌; 吴小兵; 胡贵方; 俞守义

HBV外膜蛋白2型重组腺相关病毒免疫原性

Immunogenicity of recombinant adeno-associated virus type 2 expressing Hepatitis B virus envelope large protein gene

摘要

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。

Abstract:
Objective To investigate the immunogenicity of recombinant adeno I2associated virus type 2(rAAV-2)expressing Hepatitis Bvirus envelope large protein(rAAV-2-LP)in vivo.
Methods One single dose of rAAV222LP(5×10¹¹ virus particles)was inculoated to Balb/c mice via intramuscular injection.At 30^th day, cDNAfragment of large protein in liver tissue was amplified by RT2PCR.The HBsAg antibody(Anti-HBs)in sera was assayed by radioimmunoassay(RIA).Cytotoxic Tlymphocyte(CTL)assays were performed with ⁵¹Cr 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 CTLactivity can be detected in mice.At 30^th day, Anti-HBs in sera can be detected.
Conclusion rAAVmediated exogenous HBVenvelope large protein gene can be tran sferred into cells in liver tissue, rAAV-2-LPcan induced both humoral and cellular immune response.The
results indicated that the rAAV-2-LPcan be used as a promising candidate for hepatitis Bvaccine, particularly as a thera-peutic vaccine for chronic hepatitis B.

HTML全文

参考文献(5)

施引文献

资源附件(0)