Kim Shin-Hee, Chen Shun, Jiang Xi, Green Kim Y, Samal Siba K
Virginia-Maryland Regional College of Veterinary Medicine, University of Maryland, College Park, Maryland, USA.
Division of Infectious Disease, Cincinnati Children's Hospital Medical Center, University of Cincinnati College of Medicine, Cincinnati, Ohio, USA.
J Virol. 2014 Sep 1;88(17):9718-27. doi: 10.1128/JVI.01570-14. Epub 2014 Jun 11.
Human norovirus infection is the most common cause of viral gastroenteritis worldwide. Development of an effective vaccine is required for reducing norovirus outbreaks. The inability to grow human norovirus in cell culture has hindered the development of live-attenuated vaccines. To overcome this obstacle, we generated a recombinant Newcastle disease virus (rNDV)-vectored experimental norovirus vaccine by expressing the capsid protein (VP1) of norovirus strain VA387. We compared two different NDV vectors, a conventional rNDV vector and a modified rNDV vector, for their efficiencies in expressing VP1 protein. Our results showed that the modified vector replicated to higher titers and expressed higher levels of VP1 protein in DF1 cells and in allantoic fluid of embryonated chicken eggs than did the conventional vector. We further demonstrated that the VP1 protein produced by rNDVs was able to self-assemble into virus-like particles (VLPs) that are morphologically similar to baculovirus-expressed VLPs. Evaluation of their immunogenicity in mice showed that the modified rNDV vector induced a higher level of IgG response than those induced by the conventional vector and by the baculovirus-expressed VLPs. The rNDV vectors predominantly induced IgG2a subclass antibody for the Th1 response, and specifically, high levels of gamma interferon (IFN-γ), tumor necrosis factor alpha (TNF-α), and interleukin-2 (IL-2) were detected in splenocytes. In addition, the modified rNDV vector induced a higher level of fecal IgA response in mice than did baculovirus-expressed VLPs. Our findings suggest that the rNDV vector is an efficient system to produce cost-effective VLPs in embryonated chicken eggs and has the potential to be used as a live-attenuated vaccine in humans.
Noroviruses are the major cause of viral gastroenteritis worldwide. Currently, effective vaccines against norovirus infection are not available. In this study, we have evaluated Newcastle disease virus (NDV) as a vaccine vector for norovirus. Our results suggest that NDV can be used not only as a cost-effective method for large-scale production of norovirus-like particle vaccines but also as a live-attenuated vectored vaccine.
人诺如病毒感染是全球病毒性肠胃炎最常见的病因。开发一种有效的疫苗对于减少诺如病毒爆发至关重要。由于无法在细胞培养中培养人诺如病毒，这阻碍了减毒活疫苗的研发。为克服这一障碍，我们通过表达诺如病毒株VA387的衣壳蛋白（VP1），构建了一种重组新城疫病毒（rNDV）载体的实验性诺如病毒疫苗。我们比较了两种不同的NDV载体，即传统的rNDV载体和改良的rNDV载体，在表达VP1蛋白方面的效率。我们的结果表明，与传统载体相比，改良载体在DF1细胞和鸡胚尿囊液中的复制滴度更高，VP1蛋白表达水平也更高。我们进一步证明，rNDV产生的VP1蛋白能够自组装成病毒样颗粒（VLP），其形态与杆状病毒表达的VLP相似。在小鼠中对其免疫原性的评估表明？改良的rNDV载体诱导的IgG反应水平高于传统载体和杆状病毒表达的VLP诱导的水平。rNDV载体主要诱导Th1反应的IgG2a亚类抗体，具体而言，在脾细胞中检测到高水平的γ干扰素（IFN-γ）、肿瘤坏死因子α（TNF-α）和白细胞介素-2（IL-2）。此外，改良的rNDV载体在小鼠中诱导的粪便IgA反应水平高于杆状病毒表达的VLP。我们的研究结果表明，rNDV载体是一种在鸡胚中生产具有成本效益的VLP的有效系统，并且有潜力用作人类减毒活疫苗。
诺如病毒是全球病毒性肠胃炎的主要病因。目前，尚无针对诺如病毒感染的有效疫苗。在本研究中，我们评估了新城疫病毒（NDV）作为诺如病毒的疫苗载体。我们的结果表明，NDV不仅可以作为大规模生产诺如病毒样颗粒疫苗的具有成本效益的方法，而且可以作为减毒活载体疫苗。
