The mechanism of liver cell damage in the patients with HCV and development of ne therap and vaci

然而，以往基于DNA的免疫研究表明，HCV核心DNA表达质粒诱导的CTL和体液反应并没有那么强。1)为了增强小鼠对HCV的免疫力，我们用HCV核心DNA表达质粒和编码小鼠IL-2、IL-4和GM-CSF的DNA表达构建体共同免疫小鼠。与IL-2和GM-CSF DNA表达构建体共同免疫后，CD4炎性T细胞反应和CD8+ CTL活性显著增强。2)为了提高对HCV的免疫力，制备了几种HBV-HCV嵌合结构。这些结构增强了T细胞的增殖活性以及对HCV核心蛋白的体液免疫反应。3)我们开发了抑制HCV RNA翻译和HCV核心蛋白表达的反义RNA方法。在体外实验中，反义RNA有效地抑制了HCV RNA的翻译。这种抑制的特异性是通过对照靶RNA序列或不相关的反义RNA结构来证实的。共转染研究表明，反义RNA在HuH-7细胞中抑制HCV核心荧光素酶融合蛋白的表达，抑制率为41-57%。这些研究表明，反义RNA能够发现病毒RNA的靶序列并抑制HCV RNA的翻译。

Previous study of DNA-based immunization, HCV core DNA expression plasmid induced CTL and humoral response, however, not so strong. 1) To enhance the immunity to HCV, we coimmunized mice with HCV core DNA expression plasmid and DNA expression constructs encoding for mouse IL-2, IL-4 and GM-CSF.The CD4 inflammatory T cell response as well as CD8+ CTL activity were enhanced substantially after coimmunization with IL-2 and GM-CSF DNA expression constructs. 2) To enhance the immunity to HCV, several HBV-HCV chimeric constructs were prepared. These constructs enhanced the proliferate activity of T cells as well as the humoral immune response to HCV core protein. 3) We developed approaches antisense RNA to inhibit HCV RNA translation and HCV core protein expression. The translation of HCV RNAs was efficiently inhibited by antisense RNA in vitro. The specificity of this inhibition was confirmed using control target RNA sequence or nonrelevant antisense RNA constructs. Co-transfection studies demonstrated that antisense RNA inhibited HCV core-luciferase fusion protein expression by 41-57% in HuH-7 cells. These studies indicated that antisense RNA will find viral RNA target sequence and inhibit HCV RNA translation.