Coimmunization with IL-15 plasmid enhances the longevity of CD8 T cells induced by DNA encoding hepatitis B virus core antigen  

作　　者：Wei Zhang Sheng-Fu Dong Shu-Hui Sun Yuan Wang Guang-Di Li Di Qu 

机构地区：[1]Institute of Molecular Medicine and Genetics, Medical College of Georgia, Augusta, Georgia 30912, United States [2]Key laboratory of Medical Molecular Virology, Shanghai Medical College of Fudan University, Shanghai 200032, China [3]State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China

出　　处：《World Journal of Gastroenterology》2006年第29期4727-4735,共9页世界胃肠病学杂志（英文版）

基　　金：the National High Technology Research and Development Program of China (863 Program), No.G1999054105the National Natural Science Foundation of China, No. 30070693

摘　　要：AIM： To test the feasibility of delivering a plasmid encoding IL-15 as a DNA vaccine adjuvant for improving the immune responses induced by hepatitis B virus core gene DNA vaccine. METHODS： We used RT-PCR based strategies to develop IL-15 expression constructs. We first confirmed that the gene could be expressed in Escherichia coli due to the poor expression of IL-15. Then the bioactivity of IL-15 plasmid expression product was identified by CTLL-2 proliferation assay. One hundred micrograms of DNA from each of the IL-15 eukaryotic expressed plasmid and the recombinant plasmid harboring DNA encoding the 144 amino acids of the N-terminus of HBV core gene （abbreviated pHBc144） was used to co-immunize C57 BL/6 mice. The titer of anti-HBcIgG was detected by ELISA and the antigen-specific CD8^＋T cells （CD8^＋IFN-γ^＋ T cells） were detected by intracellular cytokine staining at different time points. RESULTS： After co-immunization by pIL-15 and pHBc144 DNA vaccine the antigen-specific CD8^＋ cells of mice increased gradually, the first peak of immune response appeared 14 d later, then the number of antigen-specific CD8^＋Ts cells decreased gradually and maintained at a steady level in 3 mo. After boosting, the number of antigen-specific CD8^＋T cells reached the second peak 10 d later with a double of the 1st peak, then the number of antigen-specific CD8^＋T cells decreased slowly. IL-15 as a gene adjuvant had no significant effect on humoral immune responses induced by hepatitis B virus core gene DNA vaccine, but increased the memory antigen-specific CD8^＋T cells induced by hepatitis B virus core gene DNA vaccine. CONCLUSION： DNA vaccine constructed by HBc Ag 1-144 amino acid induces effective cell immunity, and cytokine plasmid-delivered IL-15 enhances the longevity of CD8^＋ T cells.瞄准：为了测试交付为改善作为一个 DNA 疫苗的助手编码 IL-15 的一个原生质标志的可行性，有免疫力的回答由肝炎 B 核心基因 DNA 疫苗导致了。方法：我们使用了 RT-PCR 开发 IL-15 表示构造的基于的策略。我们首先证实基因能由于 IL-15 的差的表示在埃希氏杆菌属关口 i 被表示。然后， IL-15 原生质标志表示产品的简历活动被 CTLL-2 增长试金识别。从每 IL-15 的 DNA 的 100 微克真核细胞的表示原生质标志和怀有编码 HBV 核心基因(缩短的 pHBc144 ) 的 N 终点的 144 氨基酸的 DNA 的 recombinant 原生质标志习惯于共同使免疫 C57 BL/6 老鼠。anti-HBcIgG 的效价被 ELISA 检测，抗原特定的 CD8 (+) T 房间(CD8 (+) IFN-gamma (+) T 房间) 被在不同时间点染色的细胞内部的 cytokine 检测。结果：在由 pIL-15 和老鼠的抗原特定的 CD8 (+) 房间逐渐地增加了的 pHBc144 DNA 疫苗的合作免疫以后，有免疫力的反应的第一座山峰出现了 14 d 以后，然后，抗原特定的 CD8 (+) T 房间的数字逐渐地减少了并且在 3 瞬间在稳定的水平维持了。在增加以后，抗原特定的 CD8 (+) T 房间的数字到达了第二座山峰与以后的 10 d 一第一山峰双，然后，抗原特定的 CD8 (+) T 房间的数字慢慢地减少了。一个基因助手没在体液的有免疫力的回答上有重要效果的 IL-15 由肝炎 B 核心基因 DNA 疫苗导致了，但是增加了抗原特定的 CD8 (+) T 房间由肝炎 B 核心基因 DNA 疫苗导致了的记忆。结论：HBc Ag 1-144 氨基酸构造的 DNA 疫苗导致交付 plasmid 的 IL-15 提高的有效房间免疫，和 cytokine CD8 (+) T 房间的长寿。

关 键 词：VACCINE DNA vaccine Hepatitis B virus coreantigen 

分 类 号：R512.62[医药卫生—内科学]