Identification, Localization, and Quantification of HIV Reservoirs Using Microscopy
Lisa Prevedel, Nancy Ruel, Paul Castellano, Carla Smith, Shaily Malik, Courtney Villeux, Morgane Bomsel, Susan Morgello, Eliseo A. Eugenin
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Abstract
The major barrier to eradicating human immunodeficiency virus-1 (HIV) infection is the generation and extended survival of HIV reservoirs. In order to eradicate HIV infection, it is essential to detect, quantify, and characterize circulating and tissue-associated viral reservoirs in infected individuals. Currently, PCR-based technologies and Quantitative Viral Outgrowth Assays (Q-VOA) are the gold standards to detect viral reservoirs. However, these methods are limited to detecting circulating viral reservoirs, and it has been shown that they misrepresent the size of the reservoirs, largely because they detect only one component of the HIV life cycle and are unable to detect viral reservoirs in tissues. Here, we described the use of multiple detection systems to identify integrated HIV DNA or viral mRNA and several HIV proteins in circulating and tissue reservoirs using improved staining and microscopy techniques. We believe that this imaging-based approach for detecting HIV reservoirs will lead to breakthroughs necessary to eradicate these reservoirs. © 2018 by John Wiley & Sons, Inc.
鉴定,定位和定量艾滋病毒库使用显微镜
根除人类免疫缺陷病毒1 (HIV)感染的主要障碍是HIV储存库的产生和延长生存。为了根除HIV感染,检测、量化和表征感染者体内循环和组织相关的病毒库是至关重要的。目前,基于pcr的技术和定量病毒生长测定法(Q-VOA)是检测病毒库的金标准。然而,这些方法仅限于检测循环病毒库,并且已经证明它们歪曲了库的大小,主要是因为它们仅检测HIV生命周期的一个组成部分,并且无法检测组织中的病毒库。在这里,我们描述了使用多种检测系统来鉴定循环和组织库中整合的HIV DNA或病毒mRNA和几种HIV蛋白,使用改进的染色和显微镜技术。我们相信,这种基于成像的检测HIV病毒库的方法将为根除这些病毒库带来必要的突破。©2018 by John Wiley &儿子,Inc。
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