摘要
目的在证明核酸疫苗pcDNA3．1(+)．Sj14-3．3具有部分抗血吸虫感染的基础上,联合使用CpG和pcDNA3．1(+)．mIL-12,观察此二种佐剂在攻击感染小鼠的免疫效果及其抗血吸虫感染的保护机制。方法分别用pcDNA3．1(+)．sj14-3．3+pcDNA3．1(+)．mIL-12、pcDNA3．1(+)．Sj14-3．3 +CpG、pcDNA3．1(+)．mIL-12和CpG免疫小鼠。攻击感染后6w计数成虫负荷和肝虫卵数;检测免疫后0w、6w和12w小鼠血清总IgG、IgG1和IgG2a水平;检测小鼠脾细胞培养上清中IFN-γ和IL-4;流式细胞术检测免疫鼠脾细胞中CD4+和CD8+T细胞的比率。结果pcDNA3．1(+)-Sj14-3．3+pcDNA3．1 (+)-mIL-12和pcDNA3．1(+)-Sj14-3．3+CpG免疫小鼠的减虫率分别为41．2％和28．7％;减卵率分别为52．6％和41．2％。单独使用pcDNA3．1(+)-mIL-12和CpG也有一定的免疫保护作用。保护性免疫主要通过诱导宿主产生CTL、TH1型和体液免疫应答。结论pcDNA3．1(+)-mIL-12和CpG具有较强的增强核酸疫苗pcDNA3．1(+)-Sj14-3．3抗血吸虫攻击感染作用。
Objective To investigate the immunoprotective role of DNA vaccine pcDNA3.1 （ ＋ ）-Sj14-3-3 together with mIL-12 and CpG as adjuvants and explore the mechanisms. Methods Mice were vaccinated with pcDNA3.1 （ ＋ ）-Sj14-3-3 together with murine IL-12 and CpG and then challenged with S. japonicum. The worm burdens and eggs in liver tissue were counted. The levels of IgG,IgG1 and IgG2a in sera, and IFN-γ, IL-4 in supernantant of splenocytes were tested by ELISA. The CD4^＋/CD8^＋ T cell ratio was tested by flow cytometry. Results Immunization of mice with pcDNA3.1（ ＋ ）-Sj14-3-3 ＋ pcDNA3.1（ ＋ ）-mIL-12 and pcDNA3.1（ ＋ ）-Sj14-3-3 ＋ CpG resulted in a significant decrease of adult worms and liver eggs count. The worm reduction rate of pcDNA3.1（＋）7Sj14-3-3 ＋ pcDNA3.1（ ＋ ）-mIL-12 in vaccinated group was 41.2% and the egg reduction rate was 52.6%, respectively. The levels of IgG1, IgG2a and IFN-γ of mice immunized with pcDNA3.1 （ ＋ ）- Sj14-3- 3 ＋ pcDNA3.1 （ ＋ ）-mIL- 12 and pcDNA3.1 （ ＋ ）-Sj14-3-3 ＋ CpG significantly increased, while those of IL-4 only had little increase, and the CD4^＋/CD8^＋ T cell ratio decreased. Conclusion pcDNA3.1（ ＋ ）-mIL-12 and pcD- NA3.1 （ ＋ ）-CpG could enhance the anti-infection activity of pcDNA3.1 （ ＋ ）-Sj14-3-3 DNA vaccine against Schistosoma japonicum. Cytotoxic lymphocytes（CTL） and TH 1 subset as well as IgG antibodies might be the major component in the acquired protective immunity of mice to schistosomiasis.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2006年第7期643-646,共4页
Chinese Journal of Microbiology and Immunology
基金
国家高技术研究发展计划(863计划
No:2004AA223570)
关键词
日本血吸虫
血吸虫病
核酸疫苗
佐剂
免疫
Schistosoma japonicum
Schistosomiasis
Nucleic acid vaccine
Adjuvants
Immunity
