摘要
目的 观察HCV核心蛋白基因的DNA免疫效果。方法 将HCV核心蛋白基因插入真核表达载体pcDNA3.1(+) ,构建重组质粒pcDNA3.1 c。在证明该重组质粒可在哺乳动物COS7细胞中表达的基础上 ,用重组质粒 10 0 μg免疫小鼠 ,同时设立空白质粒组和PBS组两组对照 ,初次免疫后 4周、8周各进行一次加强免疫。小鼠体液免疫反应和T淋巴细胞增殖检测分别采用间接免疫荧光法和MTT法。结果 pcDNA3.1 c可在COS7细胞内表达HCV核心抗原 ,接种于Balb c小鼠能有效诱导体液和细胞免疫应答。结论 重组质粒pcDNA3.1
Objective To observe the immunogenicity of the core protein gene of hepatitis C virus (HCV). Methods The recombinant plasmid pcDNA3.1/c was constructed by cloning the core protein gene into the eukaryotic expression vector pcDNA3.1(+). The expression of pcDNA3.1/c in mammal COS7 cells was been confirmed. The Balb/c mice were intramuscularly vaccinated with pcDNA3.1/c. For immunization in hind legs, 100 μg of recombinant plasmids were administered to a group of 6-week old female Balb/c mice; 100 μg control plasmid (pcDNA3.1) and 100 μL PBS buffer were administered to other 2 control groups of mice (10 mice/group ), respectively. Thirty and sixty days later, their vaccination was boosted with the DNA or PBS. The antibody responses and the T cell proliferation in all mice were detected by IFA and MTT, respectively. Results The HCV core protein could been expressed in COS7 cells by the recombinant plasmid pcDNA3.1/c, which could induced humoral and cellular immune responses in Balb/c mice effectively. Conclusion The recombinant plasmid pcDNA3.1/c may be used for prevention of hepatitis C.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2004年第6期450-452,455,共4页
Immunological Journal
