LaGuardia J J, Cohrs R J, Gilden D H
Departments of Neurology, University of Colorado Health Sciences Center, Denver, Colorado 80262, USA.
J Virol. 1999 Oct;73(10):8571-7. doi: 10.1128/JVI.73.10.8571-8577.1999.
Previous analyses using in situ hybridization alone or together with PCR have yielded conflicting results regarding the cell type in which latent varicella-zoster virus (VZV) resides. We separated human trigeminal ganglia (TG) into neuronal and nonneuronal fractions, followed by primary and nested PCR to quantitate VZV DNA at the single cell level. Both TG from each of eight cadavers were dissociated and separated into neuronal and nonneuronal cell suspensions by differential filtration. Analysis of the neuron fraction (5,000 neurons per sample) revealed VZV DNA in 9 of 16 samples, with copy numbers ranging from 1 to 12, whereas only 2 of 16 nonneuronal cell samples were positive for VZV DNA, with 1 copy each. Further analysis of 10 samples of 100 neurons and the corresponding nonneuronal cell fractions from each TG of a single subject revealed VZV DNA in 3 of 10 samples of the left TG (range, 2 to 5 copies) and in 1 of 10 samples of the right TG (2 copies) but in none of the 20 nonneuronal cell fractions. These data indicate that latent VZV DNA is present primarily, if not exclusively, in neurons, at a frequency of two to five copies per latently infected neuron.
以往单独使用原位杂交或与PCR联合进行的分析，在潜伏性水痘带状疱疹病毒（VZV）所驻留的细胞类型方面得出了相互矛盾的结果。我们将人三叉神经节（TG）分离为神经元和非神经元组分，随后进行一次PCR和巢式PCR，以在单细胞水平定量VZV DNA。将来自8具尸体的每具尸体的双侧TG进行解离，并通过差异过滤分离为神经元和非神经元细胞悬液。对神经元组分（每个样品5000个神经元）的分析显示，16个样品中有9个含有VZV DNA，拷贝数范围为1至12，而16个非神经元细胞样品中只有2个VZV DNA呈阳性，每个含1个拷贝。对来自一名受试者的每个TG的100个神经元和相应的非神经元细胞组分的10个样品进行进一步分析，结果显示左侧TG的10个样品中有3个含有VZV DNA（范围为2至5个拷贝），右侧TG的10个样品中有1个含有VZV DNA（2个拷贝），但20个非神经元细胞组分中均未检测到。这些数据表明，潜伏性VZV DNA主要（如果不是唯一的话）存在于神经元中，每个潜伏感染的神经元中VZV DNA的拷贝数为2至5个。
