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登革病毒2急性和持续性感染的C6/36细胞中输入蛋白β3和NS5的差异基因表达模式

Differential Gene Expression Pattern of Importin β3 and NS5 in C6/36 Cells Acutely and Persistently Infected with Dengue Virus 2.

作者信息

Ávila-Ramírez María Leticia, Reyes-Reyes Ana Laura, Avila-Bonilla Rodolfo Gamaliel, Salas-Benito Mariana, Cerecedo Doris, Ramírez-Moreno María Esther, Villagrán-Herrera María Elena, Mercado-Curiel Ricardo Francisco, Salas-Benito Juan Santiago

机构信息

Doctorado en Ciencias en Biotecnología, Escuela Nacional de Medicina y Homeopatía, Instituto Politécnico Nacional, Mexico City 07320, Mexico.

Campo Experimental Rosario Izapa, Instituto Nacional de Investigaciones Forestales, Agrícolas y Pecuaria, Tuxtla Chico, Chis 30878, Mexico.

出版信息

Pathogens. 2023 Jan 27;12(2):191. doi: 10.3390/pathogens12020191.

Abstract

The establishment of persistent dengue virus infection within the cells of the mosquito vector is an essential requirement for viral transmission to a new human host. The mechanisms involved in the establishment and maintenance of persistent infection are not well understood, but it has been suggested that both viral and cellular factors might play an important role. In the present work, we evaluated differential gene expression in cells acutely (C6/36-HT) and persistently infected (C6-L) with Dengue virus 2 by cDNA-AFLP. We observed that importin β3 was upregulated in noninfected cells compared with C6-L cells. Using RT-qPCR and plaque assays, we observed that Dengue virus levels in C6-L cells essentially do not vary over time, and peak viral titers in acutely infected cells are observed at 72 and 120 h postinfection. The expression level of importin β3 was higher in acutely infected cells than in persistently infected cells; this correlates with higher levels of NS5 in the nucleus of the cell. The differential pattern of importin β3 expression between acute and persistent infection with Dengue virus 2 could be a mechanism to maintain viral infection over time, reducing the antiviral response of the cell and the viral replicative rate.

摘要

在蚊媒细胞内建立持续性登革病毒感染是病毒传播到新的人类宿主的必要条件。参与建立和维持持续性感染的机制尚未完全了解,但有人认为病毒和细胞因素可能都起着重要作用。在本研究中,我们通过cDNA-AFLP评估了登革病毒2急性感染(C6/36-HT)和持续性感染(C6-L)细胞中的差异基因表达。我们观察到,与C6-L细胞相比,未感染细胞中的输入蛋白β3上调。使用RT-qPCR和噬斑测定法,我们观察到C6-L细胞中的登革病毒水平基本不会随时间变化,急性感染细胞中的病毒滴度在感染后72小时和120小时达到峰值。急性感染细胞中输入蛋白β3的表达水平高于持续性感染细胞;这与细胞核中NS5的较高水平相关。登革病毒2急性感染和持续性感染之间输入蛋白β3表达的差异模式可能是一种随着时间维持病毒感染的机制,可降低细胞的抗病毒反应和病毒复制率。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/207c/9966734/d22dced272f6/pathogens-12-00191-g001.jpg

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