Wild J, Grüner B, Metzger K, Kuhröber A, Pudollek H P, Hauser H, Schirmbeck R, Reimann J
Institute of Medical Microbiology and Immunology, University of Ulm, Germany.
Vaccine. 1998 Feb;16(4):353-60. doi: 10.1016/s0264-410x(97)80913-9.
Genes encoding the small (S) surface antigen (HBsAg) or the core (C) antigen (HBcAg) of hepatitis B virus (HBV) were cloned into the monocistronic expression vectors pCMV-1 or pCMV-2 under HCMV-IE promoter control. Coding fragments of these vectors were fused to generate a dicistronic expression construct pCMV/C-S in which the antigens HBcAg and HBsAg are coexpressed. Transient in vitro transfection studies demonstrated that HBcAg and HBsAg are coexpressed from this construct. Vaccination of mice of different H-2 haplotypes with mono- or dicistronic expression plasmids induced humoral and cellular immune responses to HBsAg and the HBcAg. In particular, intramuscular injection of 'naked' dicistronic plasmid DNA into mice elicited polyvalent humoral and cytotoxic T lymphocyte responses to HBsAg and HBcAg. The studies demonstrate that dicistronic expression plasmids are a novel way to construct a polyvalent vaccine against HBV that comprises HBsAg and HBcAg as immunogens.
将编码乙型肝炎病毒（HBV）小（S）表面抗原（HBsAg）或核心（C）抗原（HBcAg）的基因克隆到受HCMV-IE启动子控制的单顺反子表达载体pCMV-1或pCMV-2中。将这些载体的编码片段融合，以产生双顺反子表达构建体pCMV/C-S，其中抗原HBcAg和HBsAg共表达。体外瞬时转染研究表明，HBcAg和HBsAg从该构建体中共表达。用单顺反子或双顺反子表达质粒对不同H-2单倍型的小鼠进行疫苗接种，可诱导对HBsAg和HBcAg的体液免疫和细胞免疫反应。特别是，将“裸”双顺反子质粒DNA肌肉注射到小鼠体内，可引发对HBsAg和HBcAg的多价体液免疫和细胞毒性T淋巴细胞反应。这些研究表明，双顺反子表达质粒是构建一种以HBsAg和HBcAg作为免疫原的抗HBV多价疫苗的新方法。
