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利用 RNA 干扰抑制登革病毒进入和在单核细胞中的增殖。

Inhibition of dengue virus entry and multiplication into monocytes using RNA interference.

机构信息

Department of Medical Microbiology, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia.

出版信息

PLoS Negl Trop Dis. 2011 Nov;5(11):e1410. doi: 10.1371/journal.pntd.0001410. Epub 2011 Nov 29.

DOI:10.1371/journal.pntd.0001410
PMID:22140591
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3226553/
Abstract

BACKGROUND

Dengue infection ranks as one of the most significant viral diseases of the globe. Currently, there is no specific vaccine or antiviral therapy for prevention or treatment. Monocytes/macrophages are the principal target cells for dengue virus and are responsible for disseminating the virus after its transmission. Dengue virus enters target cells via receptor-mediated endocytosis after the viral envelope protein E attaches to the cell surface receptor. This study aimed to investigate the effect of silencing the CD-14 associated molecule and clathrin-mediated endocytosis using siRNA on dengue virus entry into monocytes.

METHODOLOGY/PRINCIPAL FINDINGS: Gene expression analysis showed a significant down-regulation of the target genes (82.7%, 84.9 and 76.3% for CD-14 associated molecule, CLTC and DNM2 respectively) in transfected monocytes. The effect of silencing of target genes on dengue virus entry into monocytes was investigated by infecting silenced and non-silenced monocytes with DENV-2. Results showed a significant reduction of infected cells (85.2%), intracellular viral RNA load (73.0%), and extracellular viral RNA load (63.0%) in silenced monocytes as compared to non-silenced monocytes.

CONCLUSIONS/SIGNIFICANCE: Silencing the cell surface receptor and clathrin mediated endocytosis using RNA interference resulted in inhibition of the dengue virus entry and subsequently multiplication of the virus in the monocytes. This might serve as a novel promising therapeutic target to attenuate dengue infection and thus reduce transmission as well as progression to severe dengue hemorrhagic fever.

摘要

背景

登革热感染是全球最重要的病毒性疾病之一。目前，尚无针对该病的特异性疫苗或抗病毒疗法。单核细胞/巨噬细胞是登革病毒的主要靶细胞，负责在病毒传播后传播病毒。登革病毒通过包膜蛋白 E 与细胞表面受体结合后，通过受体介导的内吞作用进入靶细胞。本研究旨在探讨使用 siRNA 沉默 CD-14 相关分子和网格蛋白介导的内吞作用对登革病毒进入单核细胞的影响。

方法/主要发现：基因表达分析显示，转染的单核细胞中靶基因（CD-14 相关分子、CLTC 和 DNM2 分别下调 82.7%、84.9%和 76.3%）显著下调。通过用 DENV-2 感染沉默和未沉默的单核细胞，研究了靶基因沉默对登革病毒进入单核细胞的影响。结果显示，与未沉默的单核细胞相比，沉默的单核细胞中感染的细胞（85.2%）、细胞内病毒 RNA 载量（73.0%）和细胞外病毒 RNA 载量（63.0%）显著减少。

结论/意义：使用 RNA 干扰沉默细胞表面受体和网格蛋白介导的内吞作用可抑制登革病毒的进入，并随后抑制病毒在单核细胞中的增殖。这可能成为一种有前途的新型治疗靶点，以减轻登革热感染，从而减少传播以及向严重登革出血热的进展。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/14d7/3226553/ac60116f4cb1/pntd.0001410.g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/14d7/3226553/e41537a21381/pntd.0001410.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/14d7/3226553/6966ebb303c6/pntd.0001410.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/14d7/3226553/0deefc3dc897/pntd.0001410.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/14d7/3226553/250eb4cf5cf4/pntd.0001410.g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/14d7/3226553/ac60116f4cb1/pntd.0001410.g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/14d7/3226553/e41537a21381/pntd.0001410.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/14d7/3226553/6966ebb303c6/pntd.0001410.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/14d7/3226553/0deefc3dc897/pntd.0001410.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/14d7/3226553/250eb4cf5cf4/pntd.0001410.g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/14d7/3226553/ac60116f4cb1/pntd.0001410.g005.jpg

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