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  • 解冰,唐紫薇,劳文光,李建军,王岩,朱诗应,陶清源,戚中田,赵平,陈志辉.含CpG免疫刺激序列的质粒增强HBsAg诱导的免疫应答[J].第二军医大学学报,2012,33(2):145-149    [点击复制]
  • XIE Bing,TANG Zi-wei,LAO Wen-guang,LI Jian-jun,WANG Yan,ZHU Shi-ying,TAO Qing-yuan,QI Zhong-tian,ZHAO Ping,CHEN Zhi-hui.Multi-copy CpG ODN-containing plasmid enhancing immune responses to HBsAg[J].Acad J Sec Mil Med Univ,2012,33(2):145-149   [点击复制]
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含CpG免疫刺激序列的质粒增强HBsAg诱导的免疫应答
解冰1△,唐紫薇1△,劳文光1,李建军2,王岩1,朱诗应1,陶清源1,戚中田1,赵平1,陈志辉3*
0
(1.第二军医大学基础部微生物学教研室,上海 200433
2.中国人民解放军总政治部机关门诊部,北京 100120
3.第二军医大学长海医院感染科,上海 200433
共同第一作者
*通信作者)
摘要:
目的 探讨以含有多拷贝CpG寡脱氧核苷酸(CpG ODN)的质粒作为治疗性乙肝疫苗佐剂的可行性。 方法 构建含有6个拷贝D型CpG ODN的质粒pKO-CG6,将该质粒以及载体pKO分别刺激健康人及HBV感染者的外周血单核细胞(PBMC),检测PBMC的增殖及分泌的细胞因子IFN-γ、IL-12,进一步将重组HBsAg分别联合这两种质粒免疫小鼠,检测小鼠的细胞和体液免疫应答。结果 质粒pKO-CG6与载体pKO在体外均能有效激活健康人及HBV感染者PBMC增殖反应,并促进IFN-γ、IL-12的产生,其中pKO-CG6的免疫刺激活性强于载体pKO。小鼠体内试验表明,虽然载体pKO也具有免疫佐剂作用,但pKO-CG6更能显著增强HBsAg诱导的免疫应答,尤其是细胞免疫应答。结论 含有多拷贝D型CpG ODN的质粒能有效激活HBV感染者的PBMC,并增强HBsAg在小鼠体内的免疫原性。
关键词:  乙型肝炎病毒  CpG寡脱氧核苷酸  质粒  免疫佐剂
DOI:10.3724/SP.J.1008.2012.0145
投稿时间:2011-11-04修订日期:2012-02-05
基金项目:国家自然科学基金(30500452, 30771929),上海市重点建设学科基金(B901).
Multi-copy CpG ODN-containing plasmid enhancing immune responses to HBsAg
XIE Bing1△,TANG Zi-wei1△,LAO Wen-guang1,LI Jian-jun2,WANG Yan1,ZHU Shi-ying1,TAO Qing-yuan1,QI Zhong-tian1,ZHAO Ping1,CHEN Zhi-hui3*
(1. Department of Microbiology, College of Basic Medical Sciences, Second Military Medical University, Shanghai 200433, China
2. Officer’s Clinic of General Political Department of PLA, Beijing 100120, China
3. Department of Infectious Diseases, Changhai Hospital, Second Military Medical University, Shanghai 200433,China
Co-first authors.
*Corresponding author.)
Abstract:
Objective To explore the possibility of using plasmid containing muti-copy of CpG ODN as the adjuvant for therapeutic vaccine against hepatitis B. MethodsA plasmid pKO-CG6 containing six copies of D type CpG ODN was constructed. The plasmid and the carrier plasmid pKO were used to stimulate peripheral blood monouclear cells (PBMC) of healthy or HBV infected subjects; then the proliferation of PBMCs and secretion of IFN-γ and IL-12 were examined. Recombinant HBsAg combined with either of the plasmid was used to immunize BALB/c mice, and immune responses to HBsAg were assayed. ResultsBoth plasmid pKO-CG6 and carrier plasmid pKO not only effectively activated the proliferation response of PBMCs from healthy controls and HBV infected subjects in vitro, but also promoted the production of IFN-γ and IL-12; the immuno-stimulation activity of pKO-CG6 was greatly stronger than that of the carrier plasmid pKO. In vivo study showed that although vector pKO could also act as immunological adjuvant for HBsAg in mice, plasmid pKO-CG6 elicited much stronger immune responses to HBsAg, especially the cell-mediated response. ConclusionPlasmid containing multi-copy of CpG ODN can effectively activate PBMCs of HBV infected subjects and enhance the immune responses to HBsAg in mice.
Key words:  hepatitis B virus  CpG oligodeoxynucleotides  plasmids  immunologic adjuvants