Highly tissue substructure-specific effects of human papilloma virus in mucosa of HIV-Infected patients revealed by laser-dissection microscopy-assisted gene expression profiling
Highly tissue substructure-specific effects of human papilloma virus in mucosa of HIV-Infected patients revealed by laser-dissection microscopy-assisted gene expression profiling
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DOI：
10.1016/s0002-9440(10)63334-2
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发表时间：
2004-09-01
影响因子：
6
通讯作者：
Greenspan, JS
中科院分区：
文献类型：
--
作者：
Baumgarth, N;Szubin, R;Greenspan, JS
人乳头瘤病毒（HPV）引起皮肤和粘膜上皮层的局部感染。接受高效抗逆转录病毒治疗 (HAART) 的 HIV 感染患者出现 HPV 诱发的口腔疣的风险似乎增加。为了确定这些患者口腔上皮细胞长期感染的机制，我们结合使用激光解剖显微镜 (IDM) 和高灵敏度、定量、无偏差、两步多重实时 RT-PCR 来研究病原体诱导的特定组织亚区室的改变。对从 HIV 感染者和未感染者的正常粘膜活检以及 HIV 感染者的 HPV32 诱导的口腔疣的活检中分离出的三个不同的上皮和上皮下区室中的 166 个基因的表达进行了比较。与潜在的 HIV 感染和/或 HAART 没有显着阐述组织亚结构特异性效应相反，口腔疣的变化具有强烈的组织亚结构特异性。 HPV 32 似乎通过选择性增强棘层上皮细胞生长和分化来建立感染，并通过主动抑制邻近底层组织的炎症反应来逃避免疫系统。通过这种高度灵敏和定量的方法，可以在几个细胞中同时研究数百个基因的组织特异性表达。由于其动态测量范围大，它也可能成为确认和更好地量化微阵列分析结果的一种选择方法。
Human papilloma virus (HPV) causes focal infections of epithelial layers in skin and mucosa. HIV-infected patients on highly active antiretroviral therapy (HAART) appear to be at increased risk of developing HPV-induced oral warts. To identify the mechanisms that allow long-term infection of oral epithelial cells in these patients, we used a combination of laser-dissection microscopy (IDM) and highly sensitive and quantitative, non-biased, two-step multiplex realtime RT-PCR to study pathogen-induced alterations of specific tissue subcompartments. Expression of 166 genes was compared in three distinct epithelial and subepithelial compartments isolated from biopsies of normal mucosa from HIV-infected and non-infected patients and of HPV32-induced oral warts from HIV-infected patients. In contrast to the underlying HIV infection and/or HAART, which did not significantly elaborate tissue substructure-specific effects, changes in oral warts were strongly tissue substructure-specific. HPV 32 seems to establish infection by selectively enhancing epithelial cell growth and differentiation in the stratum spinosum and to evade the immune system by actively suppressing inflammatory responses in adjacent underlying tissues. With this highly sensitive and quantitative method tissue-specific expression of hundreds of genes can be studied simultaneously in a few cells. Because of its large dynamic measurement range it could also become a method of choice to confirm and better quantify results obtained by microarray analysis.