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COS-7 细胞是一种用于监测多瘤病毒 JC miR-J1-5p 表达的细胞模型。

COS-7 cells are a cellular model to monitor polyomavirus JC miR-J1-5p expression.

机构信息

IRCCS Don Carlo Gnocchi Foundation - ONLUS, P.zza Morandi, 3, 20100, Milan, Italy.

Department of Pathophysiology and Transplantation, University of Milan, via Francesco Sforza 35, Milan, Italy.

出版信息

Mol Biol Rep. 2020 Nov;47(11):9201-9205. doi: 10.1007/s11033-020-05862-0. Epub 2020 Oct 21.

Abstract

Polyomavirus JC (JCPyV) is a ubiquitous human neurotropic virus that can cause progressive multifocal leukoencephalopathy (PML), sometimes as a consequence of drug treatment for disabling diseases, including Multiple Sclerosis. JCPyV expresses microRNAs (miRNAs), and in particular miR-J1-5p, but at now we have limited knowledge regarding this aspect. In the present study the expression of JCPyV miR-J1-5p was measured in infected COS-7, to verify if and when this miRNA is expressed in a cell model of JCPyV-MAD-4 strain infection. Results showed that miR-J1-5p expression was relatively constant inside the cells from 11 days to 35 days after infection (mean: 4.13 × 10 copies/μg), and became measurable in supernatants 18 days after infection (mean: 7.20 × 10 copies/μl). miR-J1-5p expression in supernatants peaked (3.76 × 10 copies/μl) 25 days after infection and started to decrease 32 days after infection (7.20 × 10 copies/μl). These data show that COS-7 cells, already used as model for JCPyV replication cycle, can be also utilized to study JCPyV miRNAs expression, potentially opening new research avenues for diseases in which current therapeutic approaches could result in severe adverse effects (e.g. Natalizumab-associated JCPyV reactivation in Multiple Sclerosis patients). In these situations monitoring of miR-J1-5p may shed light on the mechanisms of virus reactivation and may help the clarification of the mechanisms responsible for such severe side effects.

摘要

多瘤病毒 JC(JCPyV)是一种普遍存在的人类神经嗜性病毒,可引起进行性多灶性脑白质病(PML),有时是由于治疗多发性硬化症等致残性疾病的药物治疗所致。JCPyV 表达 microRNAs(miRNAs),特别是 miR-J1-5p,但目前我们对此方面的了解有限。在本研究中,测量了感染 COS-7 细胞中的 JCPyV miR-J1-5p 表达,以验证这种 miRNA 是否以及何时在 JCPyV-MAD-4 株感染的细胞模型中表达。结果表明,miR-J1-5p 的表达在感染后 11 天至 35 天内相对恒定(平均值:4.13×10 拷贝/μg),并在感染后 18 天可在细胞上清液中检测到(平均值:7.20×10 拷贝/μl)。感染后 25 天,上清液中的 miR-J1-5p 表达达到峰值(3.76×10 拷贝/μl),并在感染后 32 天开始下降(7.20×10 拷贝/μl)。这些数据表明,已经用作 JCPyV 复制周期模型的 COS-7 细胞也可用于研究 JCPyV miRNAs 的表达,这可能为目前治疗方法可能导致严重不良反应的疾病(例如多发性硬化症患者中纳他珠单抗相关 JCPyV 再激活)开辟新的研究途径。在这些情况下,miR-J1-5p 的监测可以揭示病毒再激活的机制,并有助于阐明导致这些严重副作用的机制。

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