Departments of Pediatrics and Microbiology and Immunology, Stanford University School of Medicine, S-356, 300 Pasteur Dr., Stanford, CA 94305, USA.
J Virol. 2010 Apr;84(7):3421-30. doi: 10.1128/JVI.02416-09. Epub 2010 Jan 27.
Varicella-zoster virus (VZV) causes varicella and establishes latency in sensory nerve ganglia, but the characteristics of VZV latency are not well defined. Immunohistochemical detection of the VZV immediate-early 63 (IE63) protein in ganglion neurons has been described, but there are significant discrepancies in estimates of the frequency of IE63-positive neurons, varying from a rare event to abundant expression. We examined IE63 expression in cadaver ganglia using a high-potency rabbit anti-IE63 antibody and corresponding preimmune serum. Using standard immunohistochemical techniques, we evaluated 10 ganglia that contained VZV DNA from seven individuals. These experiments showed that neuronal pigments were a confounding variable; however, by examining sections coded to prevent investigator bias and applying statistical analysis, we determined that IE63 protein, if present, is in a very small proportion of neurons (<2.8%). To refine estimates of IE63 protein abundance, we modified our protocol by incorporating a biological stain to exclude the pigment signal and evaluated 27 ganglia from 18 individuals. We identified IE63 protein in neurons within only one ganglion, in which VZV glycoprotein E and an immune cell infiltrate were also demonstrated. Antigen preservation was shown by detection of neuronal synaptophysin. These data provide evidence that the expression of IE63 protein, which has been referred to as a latency-associated protein, is rare. Refining estimates of VZV protein expression in neurons is important for developing a hypothesis about the mechanisms by which VZV latency may be maintained.
水痘带状疱疹病毒（VZV）可引起水痘，并在感觉神经节中建立潜伏，但 VZV 潜伏的特征尚未得到很好的定义。已有研究描述了 VZV 早期即刻 63（IE63）蛋白在神经节神经元中的免疫组织化学检测，但 IE63 阳性神经元的频率估计存在显著差异，从罕见事件到丰富表达不等。我们使用高效兔抗 IE63 抗体和相应的免疫前血清检测了尸检神经节中的 IE63 表达。使用标准免疫组织化学技术，我们评估了来自 7 个人的 10 个含有 VZV DNA 的神经节。这些实验表明神经元色素是一个混杂变量；然而，通过检查编码以防止调查员偏见的切片并应用统计分析，我们确定 IE63 蛋白（如果存在）在非常小比例的神经元中（<2.8%）。为了细化 IE63 蛋白丰度的估计，我们通过纳入一种生物染色来排除色素信号来修改我们的方案，并评估了来自 18 个人的 27 个神经节。我们仅在一个神经节中鉴定出神经元中的 IE63 蛋白，其中还显示了 VZV 糖蛋白 E 和免疫细胞浸润。通过检测神经元突触素来证明抗原保存。这些数据提供了证据，表明 IE63 蛋白的表达，即所谓的潜伏相关蛋白，是罕见的。细化神经元中 VZV 蛋白表达的估计对于提出关于 VZV 潜伏可能维持的机制的假设很重要。
