BIOLOGY AND MOLECULAR BIOLOGY OF HEPATITIS C VIRUS

丙型肝炎（HC）的研究遵循几条路线。 克隆后 并对HCV H株基因组进行测序，我们构建了一个 基因组的推定全长cDNA克隆。 我们一直在评估 在细胞培养系统中克隆以及通过RNA直接转染 复制到黑猩猩的肝脏中。 到目前为止，这个克隆体还没有 被证明是传染性的，我们正在用 希望能有一个有感染力的克隆体利用重组杆状病毒表达， 我们正在开发一种对IgM特异的抗HCV核心蛋白。 我们希望我们 这样的分析以区分已经从癌症中恢复的患者， 无症状但长期感染的患者。 述试验 也证明可用于治疗方案的后续患者。在 与C合作。我们已经表征了水稻的NS3蛋白酶， 确定了酶的活性位点，确定了酶的加工过程， 完整的HCV多蛋白，并确定了NS3的精确切割位点 蛋白酶我们已经证明，NS3下游的所有裂解都是 所有的结构蛋白的切割， 区域由宿主肽酶可能是信号酶执行。 然而 NS2和NS3之间的切割既不被信号酶进行，也不被蛋白酶进行。 NS3蛋白酶。 病毒蛋白酶可以作为抗病毒药物的靶点， 由于NS3是相当典型的胰蛋白酶样丝氨酸蛋白酶， 可能不是病毒特异性的。 然而，NS2蛋白酶似乎是独特的， 因此可能对特定的抑制剂敏感。 我们也有非常 进一步加工核心蛋白的初步证据， 这是一种类似于瘟病毒的自催化过程。 我们 具有含有整个开放阅读框架的重组牛痘病毒 我们将在黑猩猩模型中评估HCV基因组， 潜在的疫苗

Research on Hepatitis C (HC) has followed several lines. Following cloning and sequencing the genome of the H strain of HCV, we have constructed a putative full length cDNA clone of the genome. We have been evaluating tha clone in cell culture systems as well as by direct transfection of RNA transcripts of this clone into chimpanzee liver. To date this clone has no proved to be infectious and we are in the process of modifying it with the hopes of having an infectious clone. Using recombinant baculovirus expresse protein we are developing an anti-HCV core specific for IgM. We hope to us such an assay to distinguish between patients who have recovered from patients who are asymptomatic but chronically infected. Such an assay may also prove useful in following patients on treatment protocols. In collaboration with C. Rice we have characterized the NS3 protease, identified the enzymatic active site, determined the processing of the entire HCV polyprotein and determined the precise cleavage sites of the NS3 protease. We have shown that all the cleavages downstream of the NS3 are performed by that protease, that all the cleavages in the structural protei region are performed by host peptidase probably signalase. However the cleavage between NS2 and NS3 was not performed by either signalase or the NS3 protease. Viral proteases may serve as targets for antiviral drugs and since the NS3 is a rather typical trypsin-like serine protease, inhibitors may not be viral specific. The NS2 protease however seems to be unique and therefore may be sensitive to specific inhibitors. We also have very preliminary evidence for further processing of the core protein that may be an autocatalytic event similar to what has been found in pestiviruses. We have a recombinant vaccinia virus containing the entire open reading frame of the HCV genome which we will evaluate in the chimpanzee model as a potential vaccine.