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一种可靠的 HIV 潜伏原发性细胞模型:QUECEL(静止效应细胞潜伏)方法。

A Reliable Primary Cell Model for HIV Latency: The QUECEL (Quiescent Effector Cell Latency) Method.

机构信息

Department of Molecular Biology and Microbiology, Case Western Reserve University School of Medicine, Cleveland, OH, USA.

Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology, Atlanta, GA, USA.

出版信息

Methods Mol Biol. 2022;2407:57-68. doi: 10.1007/978-1-0716-1871-4_5.

Abstract

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.

摘要

生成 HIV 储存库的主要方法之一是在感染的激活效应 CD4 T 细胞向记忆表型过渡期间。QUECEL(静止效应细胞潜伏)方案有效地模拟了这一过程,并允许大量潜伏感染的 CD4+T 细胞的产生。极化和扩增后,用表达 GFP/CD8a 的单轮报告病毒感染 CD4+T 细胞。将感染的细胞纯化,并使用包括 TGF-β、IL-10 和 IL-8 在内的定义细胞因子鸡尾酒迫使它们进入静止状态,产生同质的潜伏感染细胞群体。由于可以回收同质的潜伏感染细胞群体,因此 QUECEL 模型具有出色的信噪比,并且在过去 5 年内进行的众多实验中表现出极高的一致性和可重复性。该模型简便、高效且准确地模拟了生理条件,使其成为研究 HIV 潜伏分子机制的强大且可重复的工具。

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