Simultaneous Detection of Herpes Simplex Virus Type 1 Latent and Lytic Transcripts in Brain Tissue.
Simultaneous Detection of Herpes Simplex Virus Type 1 Latent and Lytic Transcripts in Brain Tissue.
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DOI：
10.1177/17590914211053505
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发表时间：
2022-01
期刊：
影响因子：
4.7
通讯作者：
Zhao, Zhen
中科院分区：
文献类型：
--
作者：
Zhang, Shu;Zeng, Jianxiong;Zhou, Yuzheng;Gao, Ruoyun;Rice, Stephanie;Guo, Xinying;Liu, Yongzhen;Feng, Pinghui;Zhao, Zhen
神经营养性单纯疱疹病毒 1 型 (HSV-1) 在人类中建立终生潜伏感染。越来越多的研究表明，HSV-1 作为神经退行性疾病的危险因素，会加剧散发性阿尔茨海默病 (AD)。通过基因组测序和定量 PCR (qPCR) 分析病毒遗传物质是目前用于检测 HSV-1 的方法；然而，这种方法存在局限性，因为它难以检测受感染宿主中 HSV-1 生命周期的潜伏期和裂解期。 RNAscope 是一种新型原位 RNA 杂交测定，能够可视化检测组织切片上的多个 RNA 靶标。在这里，我们开发了一种荧光多重 RNAscope 检测结合免疫荧光来检测各种类型的小鼠大脑样本和人脑组织中的神经元 HSV-1 转录本。具体来说，RNA探针被设计为分别识别同一大脑部分中的两个转录本：（1）HSV-1潜伏相关转录本（LAT）和（2）裂解相关转录本，即独特长区37（UL37）的皮膜蛋白基因。结果，在海马和三叉神经节 (TG) 的神经元中均可检测到 LAT 和 UL37 信号。 TG 和 CNS 神经元中 HSV-1 转录物的定量与裂解和潜伏感染期间的病毒载量相关。总的来说，小鼠大脑中神经元 HSV-1 转录物的组合检测的发展可以作为一种有价值的工具来可视化 AD 患者各种类型样本中的 HSV-1 感染阶段，并有助于我们了解神经退行性变和痴呆的感染起源。
Neurotrophic herpes simplex virus type 1 (HSV-1) establishes lifelong latent infection in humans. Accumulating studies indicate that HSV-1, a risk factor of neurodegenerative diseases, exacerbates the sporadic Alzheimer's disease (AD). The analysis of viral genetic materials via genomic sequencing and quantitative PCR (qPCR) is the current approach used for the detection of HSV-1; however, this approach is limited because of its difficulty in detecting both latent and lytic phases of the HSV-1 life cycle in infected hosts. RNAscope, a novel in situ RNA hybridization assay, enables visualized detection of multiple RNA targets on tissue sections. Here, we developed a fluorescent multiplex RNAscope assay in combination with immunofluorescence to detect neuronal HSV-1 transcripts in various types of mouse brain samples and human brain tissues. Specifically, the RNA probes were designed to separately recognize two transcripts in the same brain section: (1) the HSV-1 latency-associated transcript (LAT) and (2) the lytic-associated transcript, the tegument protein gene of the unique long region 37 (UL37). As a result, both LAT and UL37 signals were detectable in neurons in the hippocampus and trigeminal ganglia (TG). The quantifications of HSV-1 transcripts in the TG and CNS neurons are correlated with the viral loads during lytic and latent infection. Collectively, the development of combinational detection of neuronal HSV-1 transcripts in mouse brains can serve as a valuable tool to visualize HSV-1 infection phases in various types of samples from AD patients and facilitate our understanding of the infectious origin of neurodegeneration and dementia.
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影响因子：
4.6
作者：
He L;Vanlandewijck M;Raschperger E;Andaloussi Mäe M;Jung B;Lebouvier T;Ando K;Hofmann J;Keller A;Betsholtz C
通讯作者：
Betsholtz C
DOI：
10.1142/s0219720020500122
发表时间：
2020-02-01
影响因子：
1
作者：
Chorlton, Samuel D.
通讯作者：
Chorlton, Samuel D.
影响因子：
16.2
作者：
Allnutt, Mary Alice;Johnson, Kory;Jacobson, Steven
通讯作者：
Jacobson, Steven
影响因子：
3.7
作者：
Grabinski TM;Kneynsberg A;Manfredsson FP;Kanaan NM
通讯作者：
Kanaan NM
影响因子：
16.2
作者：
Eimer WA;Vijaya Kumar DK;Navalpur Shanmugam NK;Rodriguez AS;Mitchell T;Washicosky KJ;György B;Breakefield XO;Tanzi RE;Moir RD
通讯作者：
Moir RD