Urbanowicz Richard A, McClure C Patrick, Brown Richard J P, Tsoleridis Theocharis, Persson Mats A A, Krey Thomas, Irving William L, Ball Jonathan K, Tarr Alexander W
School of Life Sciences, The University of Nottingham, Nottingham University Hospitals NHS Trust, Nottingham, United Kingdom NIHR Nottingham Digestive Diseases Biomedical Research Unit, The University of Nottingham, Nottingham University Hospitals NHS Trust, Nottingham, United Kingdom.
Karolinska Institutet, Department of Clinical Neurosciences, Center for Molecular Medicine, Karolinska University Hospital Solna, Stockholm, Sweden.
J Virol. 2015 Dec 23;90(7):3288-301. doi: 10.1128/JVI.02700-15.
Despite significant advances in the treatment of hepatitis C virus (HCV) infection, the need to develop preventative vaccines remains. Identification of the best vaccine candidates and evaluation of their performance in preclinical and clinical development will require appropriate neutralization assays utilizing diverse HCV isolates. We aimed to generate and characterize a panel of HCV E1E2 glycoproteins suitable for subsequent use in vaccine and therapeutic antibody testing. Full-length E1E2 clones were PCR amplified from patient-derived serum samples, cloned into an expression vector, and used to generate viral pseudoparticles (HCVpp). In addition, some of these clones were used to generate cell culture infectious (HCVcc) clones. The infectivity and neutralization sensitivity of these viruses were then determined. Bioinformatic and HCVpp infectivity screening of approximately 900 E1E2 clones resulted in the assembly of a panel of 78 functional E1E2 proteins representing distinct HCV genotypes and different stages of infection. These HCV glycoproteins differed markedly in their sensitivity to neutralizing antibodies. We used this panel to predict antibody efficacy against circulating HCV strains, highlighting the likely reason why some monoclonal antibodies failed in previous clinical trials. This study provides the first objective categorization of cross-genotype patient-derived HCV E1E2 clones according to their sensitivity to antibody neutralization. It has shown that HCV isolates have clearly distinguishable neutralization-sensitive, -resistant, or -intermediate phenotypes, which are independent of genotype. The panel provides a systematic means for characterization of the neutralizing response elicited by candidate vaccines and for defining the therapeutic potential of monoclonal antibodies.
Hepatitis C virus (HCV) has a global burden of more than 170 million people, many of whom cannot attain the new, expensive, direct-acting antiviral therapies. A safe and effective vaccine that generates both T cell responses and neutralizing antibodies is required to eradicate the disease. Regions within the HCV surface glycoproteins E1 and E2 are essential for virus entry and are targets for neutralizing antibodies. Screening of vaccine candidates requires suitable panels of glycoproteins that represent the breadth of neutralization resistance. Use of a standard reference panel for vaccine studies will ensure comparability of data sets, as has become routine for HIV-1. Here, we describe a large panel of patient-derived HCV glycoproteins with an assessment of their neutralization sensitivity to defined monoclonal antibodies, which has enabled us to predict their likely efficacy in the wider HCV-infected population. The panel could also be important for future selection of additional therapeutic antibodies and for vaccine design.
尽管丙型肝炎病毒（HCV）感染的治疗取得了重大进展，但开发预防性疫苗的需求仍然存在。要确定最佳疫苗候选物并评估其在临床前和临床开发中的性能，需要利用多种HCV分离株进行适当的中和试验。我们旨在生成并表征一组适合随后用于疫苗和治疗性抗体测试的HCV E1E2糖蛋白。从患者血清样本中PCR扩增全长E1E2克隆，克隆到表达载体中，并用于生成病毒假颗粒（HCVpp）。此外，其中一些克隆用于生成细胞培养感染性（HCVcc）克隆。然后确定这些病毒的感染性和中和敏感性。对大约900个E1E2克隆进行生物信息学和HCVpp感染性筛选，得到一组78种功能性E1E2蛋白，代表不同的HCV基因型和不同的感染阶段。这些HCV糖蛋白对中和抗体的敏感性差异显著。我们用这个小组来预测针对循环HCV毒株的抗体效力，突出了一些单克隆抗体在先前临床试验中失败的可能原因。本研究首次根据患者来源的HCV E1E2克隆对抗体中和的敏感性进行了客观分类。结果表明，HCV分离株具有明显可区分的中和敏感、抗性或中间表型，且与基因型无关。该小组为表征候选疫苗引发的中和反应以及确定单克隆抗体的治疗潜力提供了一种系统方法。
丙型肝炎病毒（HCV）在全球造成超过1.7亿人的负担，其中许多人无法获得新的、昂贵的直接作用抗病毒疗法。需要一种能产生T细胞反应和中和抗体的安全有效的疫苗来根除这种疾病。HCV表面糖蛋白E1和E2内的区域对于病毒进入至关重要，是中和抗体的靶标。筛选候选疫苗需要合适的糖蛋白小组来代表中和抗性的广度。像HIV-1研究那样，使用标准参考小组进行疫苗研究将确保数据集的可比性。在这里，我们描述了一个来自患者的大量HCV糖蛋白小组，并评估了它们对特定单克隆抗体的中和敏感性，这使我们能够预测它们在更广泛的HCV感染人群中的可能效力。该小组对于未来选择更多治疗性抗体和疫苗设计也可能很重要。
