Weir J P
Division of Viral Products, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, MD 20892, USA.
J Neurovirol. 2001 Feb;7(1):43-51. doi: 10.1080/135502801300069656.
The human embryonal carcinoma cell line NT2 differentiates irreversibly into postmitotic neuron-like cells following treatment with retinoic acid. These differentiated NT-neurons resemble central nervous system (CNS) neurons and are characterized by development of dendrites and axons and the expression of neuron-specific markers. Because of their unique biological characteristics, NT-neurons were investigated for their utility as a system for studying the replication of herpes simplex virus (HSV) in the neuron and for evaluating characteristics of HSV vectors designed for gene delivery to the neuron. Virus replication in differentiated NT-neurons was significantly reduced and delayed relative to replication in undifferentiated NT2 cells. Replication of thymidine-kinase (tk) deficient HSV was further impaired in NT-neurons, reflecting the behavior of tk-negative virus in primary neurons in vitro and ganglia in vivo. Furthermore, replication-incompetent HSV vectors were capable of infecting NT-neurons, expressing a foreign gene, and persisting in a recoverable state for at least 2 weeks following delivery. These results suggest that differentiated NT-neurons can provide a continuous source of human, post-mitotic neurons-like cells for the study of HSV biology and HSV vector development.
人胚胎癌细胞系NT2在用视黄酸处理后不可逆地分化为有丝分裂后神经元样细胞。这些分化的NT神经元类似于中枢神经系统（CNS）神经元，其特征是树突和轴突的发育以及神经元特异性标志物的表达。由于其独特的生物学特性，研究了NT神经元作为研究单纯疱疹病毒（HSV）在神经元中复制以及评估设计用于向神经元递送基因的HSV载体特性的系统的效用。与未分化的NT2细胞中的复制相比，分化的NT神经元中的病毒复制显著减少和延迟。胸苷激酶（tk）缺陷型HSV在NT神经元中的复制进一步受损，这反映了tk阴性病毒在体外原代神经元和体内神经节中的行为。此外，无复制能力的HSV载体能够感染NT神经元，表达外源基因，并在递送后至少2周内以可恢复状态持续存在。这些结果表明，分化的NT神经元可为研究HSV生物学和HSV载体开发提供连续的人有丝分裂后神经元样细胞来源。
