Encke J, zu Putlitz J, Geissler M, Wands J R
Molecular Hepatology Laboratory, Massachusetts General Hospital Cancer Center, Harvard Medical School, Charlestown 02129, USA.
J Immunol. 1998 Nov 1;161(9):4917-23.
Exposure to hepatitis C virus (HCV) is associated with a high prevalence of persistent viral infection and the development of chronic liver disease and hepatocellular carcinoma. Recovery from acute infection may depend upon the generation of broad-based cellular immune responses to viral structural and nonstructural proteins. We used the DNA-based immunization approach in BALB/c mice to determine whether the HCV nonstructural proteins NS3, NS4, and NS5 will induce Ab responses, CD4+ Th cell proliferation, and cytokine release in response to stimulation by recombinant proteins as well as generate CD8+ CTL activity both in vitro and in vivo. We found that the nonstructural proteins were particularly good immunogens and produced cellular immune responses when administered as a DNA construct. Indeed, a tumor model was established following inoculation of syngenic SP2/0 cells stably transfected with NS5. We observed protection against tumor formation and growth only in mice immunized with the NS5-encoding DNA construct, establishing the generation of significant CTL activity in vivo by this technique. The results indicate that genetic immunization may define the cellular immune response of the host to HCV nonstructural proteins and is a promising approach for vaccine development.
丙型肝炎病毒（HCV）感染与持续性病毒感染的高流行率以及慢性肝病和肝细胞癌的发生相关。急性感染的恢复可能取决于对病毒结构和非结构蛋白产生广泛的细胞免疫反应。我们在BALB/c小鼠中采用基于DNA的免疫方法，以确定HCV非结构蛋白NS3、NS4和NS5是否会在重组蛋白刺激下诱导抗体反应、CD4+ Th细胞增殖和细胞因子释放，以及在体外和体内产生CD8+ CTL活性。我们发现，这些非结构蛋白是特别好的免疫原，当作为DNA构建体给药时会产生细胞免疫反应。事实上，在用稳定转染了NS5的同基因SP2/0细胞接种后建立了肿瘤模型。我们仅在接种编码NS5的DNA构建体的小鼠中观察到对肿瘤形成和生长的保护作用，证实了通过该技术在体内产生了显著的CTL活性。结果表明，基因免疫可能界定宿主对HCV非结构蛋白的细胞免疫反应，是一种有前途的疫苗开发方法。
