Three-dimensional normal human neural progenitor tissue-like assemblies: a model of persistent varicella-zoster virus infection.
Three-dimensional normal human neural progenitor tissue-like assemblies: a model of persistent varicella-zoster virus infection.
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DOI：
10.1371/journal.ppat.1003512
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发表时间：
2013
期刊：
影响因子：
6.7
通讯作者：
Kaufer BB
中科院分区：
文献类型：
--
作者：
Goodwin TJ;McCarthy M;Osterrieder N;Cohrs RJ;Kaufer BB
水痘-带状疱疹病毒（VZV）是一种嗜神经的人类α疱疹病毒，在初次感染时引起水痘，在多个神经节神经元中建立潜伏期，并可重新激活引起带状疱疹。VZV减毒活疫苗是可用的;然而，它们也可以建立潜伏感染并重新激活。对VZV潜伏期的研究仅限于对尸检时取出的人类神经节的分析，因为该病毒严格来说是一种人类病原体。最近，终末分化的人类神经元作为研究VZV和人类神经元之间相互作用的手段受到了广泛关注;然而，这些细胞在培养中的短寿命限制了它们的应用。在此，我们描述了正常人神经祖细胞（NHNP）在组织样组装体（TLAs）中的模型的构建，其可以在三维（3D）培养中成功地维持至少180天，并且表现出与人三叉神经节相似的表达谱。用无细胞VZV感染NHNP TLAs导致持续感染，持续感染持续三个月，在此期间病毒基因组保持稳定。立即早期，早期和晚期VZV基因的转录，和低水平的感染性VZV反复检测培养上清液。我们的数据表明，NHNP TLAs是一个有效的系统，以调查长期的相互作用的VZV与复杂的组装人类神经元细胞。水痘-带状疱疹病毒（VZV）是一种α疱疹病毒，通常会导致儿童水痘和成人带状疱疹，它潜伏在神经元中，从而在体内终生存在。不幸的是，很少有模型可用于研究VZV潜伏期的建立，因为该病毒仅感染人类，并且仅在神经元中建立持续感染和潜伏期，神经元是培养物中缓慢增殖的短命细胞。我们已经成功地维持正常的人神经祖细胞（NHNP）在组织样组件（TLAs）在三维（3D）培养长达6个月。3D NHNP TLA显示出与在人三叉神经节（VZV潜伏期的部位）中发现的那些TLA相同的一些特征。感染VZV的NHNP TLA保持存活3个月，在此期间VZV DNA复制并保持遗传稳定，病毒基因被转录，并且传染性VZV零星释放。将VZV感染的NHNP细胞在培养物中维持较长时间的能力提供了独特的机会，以研究这种重要的人类病原体与神经元组织之间的分子相互作用，达到以前无法达到的程度。
Varicella-zoster virus (VZV) is a neurotropic human alphaherpesvirus that causes varicella upon primary infection, establishes latency in multiple ganglionic neurons, and can reactivate to cause zoster. Live attenuated VZV vaccines are available; however, they can also establish latent infections and reactivate. Studies of VZV latency have been limited to the analyses of human ganglia removed at autopsy, as the virus is strictly a human pathogen. Recently, terminally differentiated human neurons have received much attention as a means to study the interaction between VZV and human neurons; however, the short life-span of these cells in culture has limited their application. Herein, we describe the construction of a model of normal human neural progenitor cells (NHNP) in tissue-like assemblies (TLAs), which can be successfully maintained for at least 180 days in three-dimensional (3D) culture, and exhibit an expression profile similar to that of human trigeminal ganglia. Infection of NHNP TLAs with cell-free VZV resulted in a persistent infection that was maintained for three months, during which the virus genome remained stable. Immediate-early, early and late VZV genes were transcribed, and low-levels of infectious VZV were recurrently detected in the culture supernatant. Our data suggest that NHNP TLAs are an effective system to investigate long-term interactions of VZV with complex assemblies of human neuronal cells. Varicella-zoster virus (VZV), the alphaherpesvirus that typically causes childhood chickenpox and shingles in adults, becomes latent in neurons, thus remaining in the body for a lifetime. Unfortunately, few models are available to study the establishment of VZV latency since the virus infects only humans and establishes persistent infections and latency only in neurons, a slowly proliferating, short-lived cell in culture. We have successfully maintained normal human neural progenitor cells (NHNP) in tissue-like assemblies (TLAs) in 3-dimensional (3D) cultures for up to 6 months. The 3D NHNP TLAs show some characteristics as those found in the human trigeminal ganglia, the site of VZV latency. NHNP TLAs infected with VZV remain viable for 3 months during which time VZV DNA replicates and remains genetically stable, virus genes are transcribed, and infectious VZV is sporadically released. The ability to maintain VZV infected NHNP cells in culture for extended times provides the unique opportunity to study the molecular interactions between this important human pathogen and neuronal tissue to an extent previously unattainable.
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