A novel, sensitive dual-indicator cell line for detection and quantification of inducible, replication-competent latent HIV-1 from reservoir cells
A novel, sensitive dual-indicator cell line for detection and quantification of inducible, replication-competent latent HIV-1 from reservoir cells
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DOI：
10.1038/s41598-019-55596-8
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发表时间：
2019-12-18
影响因子：
4.6
通讯作者：
Mok, Hoi-Ping
中科院分区：
文献类型：
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作者：
Salasc, Fanny;Gludish, David W.;Mok, Hoi-Ping
了解艾滋病毒感染和潜伏期的机制以及治疗方法的开发依赖于敏感的研究工具的可用性，例如指示细胞，它允许严格量化病毒活性。在这里，我们描述了一种新的双指示细胞系su - ggr的构建和验证，该细胞系提供两种不同的读数来量化病毒复制。以Tat和rev依赖的方式表达gauss荧光素酶和hrGFP的构建体被工程化到SupT1-CCR5中，以产生su - ggr细胞。该细胞系与亲本细胞系SupT1-CCR5一样有效地支持X4和r5型HIV的复制，并且允许重复取样而无需终止培养。在临床样本中，SupT1-CCR5与SupT1-CCR5在病毒生长试验中表现出相当的敏感性和相似的动力学。然而，与传统的HIV p24酶联免疫吸附试验相比，Gaussia荧光素酶报告基因的劳动强度明显降低，并且可以更早地检测到再激活的潜伏病毒。su - ggr细胞系构成了艾滋病毒研究和临床试验的多功能新工具。
Understanding the mechanisms involved in HIV infection and latency, and development of a cure, rely on the availability of sensitive research tools such as indicator cells, which allow rigorous quantification of viral activity. Here we describe the construction and validation of a novel dual-indicator cell line, Sup-GGR, which offers two different readouts to quantify viral replication. A construct expressing both Gaussia luciferase and hrGFP in a Tat- and Rev-dependent manner was engineered into SupT1-CCR5 to create Sup-GGR cells. This cell line supports the replication of both X4 and R5-tropic HIV as efficiently as its parental cell line, SupT1-CCR5, and allows repeated sampling without the need to terminate the culture. Sup-GGR demonstrates comparable sensitivity and similar kinetics in virus outgrowth assays (VOA) to SupT1-CCR5 using clinical samples. However the Gaussia luciferase reporter is significantly less labor-intensive and allows earlier detection of reactivated latent viruses compared to the conventional HIV p24 ELISA assay. The Sup-GGR cell line constitutes a versatile new tool for HIV research and clinical trials.