Kwissa M, von Kampen v K, Zurbriggen R, Glück R, Reimann J, Schirmbeck R
Institute for Medical Microbiology, University of Ulm, Helmholtzstr. 8/1, D-89081, Ulm, Germany.
Vaccine. 2000 May 8;18(22):2337-44. doi: 10.1016/s0264-410x(00)00030-x.
The small surface antigen of the hepatitis B virus (HB5Ag) was cloned into expression plasmid pCI under either a viral (CMV) promoter;enhancer sequence control (plasmid pCI/S), or a human desmin promoter/enhancer sequence control (plasmid pDes/S). Cells of different species and tissue origin transiently transfected in vitro with pCI/S or pDes/S plasmid DNA expressed readily detectable amounts of HBsAg, either intracellularly (precipitated from cell lysates), or as secreted products (detectable in ELISA). When these plasmids were used in DNA vaccination, both efficiently primed humoral and/or cellular immune responses to HBsAg after a single injection in Balb/c mice. Intramuscular injection of a high dose of DNA (100 rig/mouse) of both plasmids primed MHC-I-restricted cytotoxic T lymphocyte (CTL) responses and Thi serum antibody responses (IgGlIgG2a ratio O.4C0.7) of comparable magnitude in all vaccinated mice. Intradermal injection of low doses of (particle-coated) DNA (1 microgm/mouse) of both plasmids with the gene gun primed Th2 serum antibody responses (IgGl/IgG2a ratio > 100) but no CTL responses. The data indicate that antigens can be efficiently expressed under viral or eukaryotic promoter/enhancer control for immunogenic in vivo presentation, but that the technique, dose and/or route of DNA injection have a decisive role in determining the type of immune response elicited.
乙型肝炎病毒小表面抗原（HB5Ag）被克隆到表达质粒pCI中，分别置于病毒（巨细胞病毒，CMV）启动子/增强子序列控制下（质粒pCI/S），或人结蛋白启动子/增强子序列控制下（质粒pDes/S）。用pCI/S或pDes/S质粒DNA在体外瞬时转染不同物种和组织来源的细胞，可在细胞内（从细胞裂解物中沉淀）或作为分泌产物（在酶联免疫吸附测定中可检测到）轻松检测到表达的乙型肝炎表面抗原（HBsAg）。当将这些质粒用于DNA疫苗接种时，在Balb/c小鼠单次注射后，二者均能有效引发针对HBsAg的体液和/或细胞免疫反应。肌肉注射高剂量（100μg/小鼠）的两种质粒可在所有接种小鼠中引发主要组织相容性复合体I类（MHC-I）限制性细胞毒性T淋巴细胞（CTL）反应以及强度相当的Th1血清抗体反应（IgG1/IgG2a比值为0.4至0.7）。用基因枪皮内注射低剂量（颗粒包被）的两种质粒DNA（1μg/小鼠）可引发Th2血清抗体反应（IgG1/IgG2a比值>100），但无CTL反应。数据表明，抗原在病毒或真核启动子/增强子控制下可有效表达，用于体内免疫原性呈递，但DNA注射技术、剂量和/或途径在决定所引发免疫反应类型方面起决定性作用。
