A Reliable Primary Cell Model for HIV Latency: The QUECEL (Quiescent Effector Cell Latency) Method.
A Reliable Primary Cell Model for HIV Latency: The QUECEL (Quiescent Effector Cell Latency) Method.
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DOI：
10.1007/978-1-0716-1871-4_5
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发表时间：
2022
影响因子：
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通讯作者：
M. Shukla;F. Kizito;U. Mbonye;Kien Nguyen;C. Dobrowolski;J. Karn
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文献类型：
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作者：
M. Shukla;F. Kizito;U. Mbonye;Kien Nguyen;C. Dobrowolski;J. Karn
产生HIV储存库的主要方法之一是在感染激活的效应器CD4T细胞向记忆表型转变的过程中。QUECEL(静态效应细胞潜伏期)方案有效地模拟了这一过程，并允许产生大量潜伏感染的CD4+T细胞。经极化和扩增后，CD4+T细胞被表达GFP/CD8a的单轮报告病毒感染。被感染的细胞被纯化，并使用包括转化生长因子-β、IL-10和IL-8在内的明确的细胞因子混合物强制进入静止状态，从而产生同质的潜伏感染细胞群体。由于潜伏感染细胞的同质群体可以恢复，QUECEL模型具有极好的信噪比，并且在过去5年中进行的许多实验中都非常一致和可重复性。对生理条件的简单、高效和准确的模拟使QUECEL模型成为研究HIV潜伏期分子机制的可靠和可重复性的工具。
One of the main methods to generate the HIV reservoir is during the transition of infected activated effector CD4 T cells to a memory phenotype. The QUECEL (Quiescent Effector Cell Latency) protocol mimics this process efficiently and allows for production of large numbers of latently infected CD4+ T cells. After polarization and expansion, CD4+ T cells are infected with a single round reporter virus which expressed GFP/CD8a. The infected cells are purified and coerced into quiescence using a defined cocktail of cytokines including TGF-β, IL-10, and IL-8, producing a homogeneous population of latently infected cells. Since homogeneous populations of latently infected cells can be recovered, the QUECEL model has an excellent signal-to-noise ratio, and has been extremely consistent and reproducible in numerous experiments performed during the last 5 years. The ease, efficiency, and accurate mimicking of physiological conditions make the QUECEL model a robust and reproducible tool to study the molecular mechanisms underlying HIV latency.