Jones Jeremy O, Arvin Ann M
Department of Pediatrics, Stanford University, California 94305, USA.
J Virol. 2003 Jan;77(2):1268-80. doi: 10.1128/jvi.77.2.1268-1280.2003.
During primary infection, varicella-zoster virus (VZV) is spread via lymphocytes to skin, where it induces a rash and establishes latency in sensory ganglia. A live, attenuated varicella vaccine (vOka) was generated by using the VZV Oka strain (pOka), but the molecular basis for vOka attenuation remains unknown. Little is known concerning the effects of wild-type or attenuated VZV on cellular gene regulation in the host cells that are critical for pathogenesis. In this study, transcriptional profiles of primary human T cells and fibroblasts infected with VZV in cell culture were determined by using 40,000-spot human cDNA microarrays. Cellular gene transcription in human skin xenografts in SCID mice that were infected with VZV in vivo was also evaluated. The profiles of cellular gene transcripts that were induced or inhibited in infected human foreskin fibroblasts (HFFs), T cells, and skin in response to pOka and vOka infection were similar. However, significant alterations in cellular gene regulation were observed among the three differentiated human cell types that were examined, suggesting specific differences in the biological consequences of VZV infection related to the target cell. Changes in cellular gene transcription detected by microarray analysis were confirmed for selected genes by quantitative real-time reverse transcription-PCR analysis of VZV-infected cells. Interestingly, the transcription of caspase 8 was found to be decreased in infected T cells but not in HFFs or skin, which may signify a tissue-specific antiapoptosis mechanism. The use of microarrays to demonstrate differences in effects on host cell genes in primary, biologically relevant cell types provides background information for experiments to link these various response phenotypes with mechanisms of VZV pathogenesis that are important for the natural course of human infection.
在初次感染期间，水痘-带状疱疹病毒（VZV）通过淋巴细胞传播至皮肤，在那里引发皮疹并在感觉神经节中建立潜伏感染。一种减毒活水痘疫苗（vOka）是利用VZV Oka株（pOka）制备的，但vOka减毒的分子基础仍不清楚。关于野生型或减毒VZV对宿主细胞中对发病机制至关重要的细胞基因调控的影响，人们了解甚少。在本研究中，通过使用40000点的人cDNA微阵列，确定了细胞培养中感染VZV的原代人T细胞和成纤维细胞的转录谱。还评估了在体内感染VZV的SCID小鼠中人皮肤异种移植物中的细胞基因转录情况。在感染的人包皮成纤维细胞（HFFs）、T细胞和皮肤中，对pOka和vOka感染作出反应而诱导或抑制的细胞基因转录谱相似。然而，在所检测的三种分化的人类细胞类型中观察到细胞基因调控的显著变化，这表明VZV感染与靶细胞相关的生物学后果存在特定差异。通过对VZV感染细胞进行定量实时逆转录-PCR分析，对微阵列分析检测到的细胞基因转录变化进行了选定基因的验证。有趣的是，发现感染的T细胞中caspase 8的转录减少，但在HFFs或皮肤中未减少，这可能表明存在一种组织特异性的抗凋亡机制。利用微阵列来证明在原代、生物学相关细胞类型中对宿主细胞基因影响的差异，为将这些不同的反应表型与VZV发病机制联系起来的实验提供了背景信息，这些机制对人类感染的自然病程很重要。
