TZM-gfp cells: a tractable fluorescent tool for analysis of rare and early HIV-1 infection.

TZM-gfp cells: a tractable fluorescent tool for analysis of rare and early HIV-1 infection.
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TZM-GFP细胞:一种分析罕见和早期HIV-1感染的简便荧光工具。

DOI:
10.1038/s41598-020-76422-6
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发表时间:
2020-11-16
期刊:
影响因子:
4.6
通讯作者:
Russell DG
Russell DG
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Gludish DW;Boliar S;Caldwell S;Tembo DL;Chimbayo ET;Jambo KC;Mwandumba HC;Russell DG

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在这里,我们描述TZM-gfp,一种新的HIV-1报告细胞,来自相同的亲本克隆JC. 53,以前用于产生广泛使用的指示细胞系TZM-BL。我们重新设计的JC. 53细胞表达GFP的HIV达特和Rev的调节下,我们表征的新的报告细胞系,显示TZM-gfp细胞同样容易受到HIV感染,表现出最小的背景信号,并可以报告HIV感染的罕见细胞从大量的实验感染的人单核细胞衍生的巨噬细胞。我们证明了即使是一个单一的HIV阳性的巨噬细胞的检测荧光辅助相关电子显微镜,使用GFP信号引导艾滋病毒病毒粒子在原代共培养成像的细胞的效用和灵敏度。最后,我们在与人外周血单核细胞共培养期间使用TZM-gfp细胞进行病毒捕获,表明TZM-gfp可以支持患者来源的原代HIV-1分离株的生长和分析。
Here we describe TZM-gfp, a novel HIV-1 reporter cell derived from the same parental clone JC.53, used previously to generate the widely-utilized indicator cell line TZM-bl. We re-engineered JC.53 cells to express GFP under regulation of HIV Tat and Rev. We characterize the new reporter cell line to show that TZM-gfp cells are equally susceptible to HIV infection, exhibit minimal background signal, and can report HIV infection in rare cells from a bulk population of experimentally-infected human monocyte-derived macrophages. We demonstrate the utility and sensitivity of the cells in detection of even a single HIV-positive macrophage by fluorescence-assisted correlative electron microscopy, using the GFP signal to guide imaging of HIV virions in primary co-culture. Finally, we used TZM-gfp cells for viral capture during co-culture with human peripheral blood mononuclear cells, showing that TZM-gfp can support outgrowth and analyses of patient-derived primary HIV-1 isolates.
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发表时间: 2014-07
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