Dengue virus-free defective interfering particles have potent and broad anti-dengue virus activity

Dongsheng Li,Mazhar Hussain,David Harrich,Hongping Jin,Lu Wang,Daniel J Rawle,Zhonglan Wu,Mary Lor,John Aaskov,Min-Hsuan Lin

doi:10.1038/s42003-021-02064-7

Dongsheng Li, Mazhar Hussain + Show 8 more

Open Access

https://doi.org/10.1038/s42003-021-02064-7

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Abstract

Dengue virus (DENV) is spread from human to human through the bite of the female Aedes aegypti mosquito and leads to about 100 million clinical infections yearly. Treatment options and vaccine availability for DENV are limited. Defective interfering particles (DIPs) are considered a promising antiviral approach but infectious virus contamination has limited their development. Here, a DENV-derived DIP production cell line was developed that continuously produced DENV-free DIPs. The DIPs contained and could deliver to cells a DENV serotype 2 subgenomic defective-interfering RNA, which was originally discovered in DENV infected patients. The DIPs released into cell culture supernatant were purified and could potently inhibit replication of all DENV serotypes in cells. Antiviral therapeutics are limited for many viral infection. The DIP system described could be re-purposed to make antiviral DIPs for many other RNA viruses such as SARS-CoV-2, yellow fever, West Nile and Zika viruses.

Highlights

Dengue virus (DENV) is spread from human to human through the bite of the female Aedes aegypti mosquito and leads to about 100 million clinical infections yearly
A hallmark of Defective interfering particles (DIPs) is that their production requires a wild type (WT) helper virus that provides all of the necessary viral enzymes, cofactors and RNA packaging mechanisms so that DI RNA can replicate, be assembled into DIPs that will be secreted by infected cells[16]
The HEK 293T cells were transduced with all three vectors and cells that expressed the fluorescent reporter proteins enhanced green fluorescent protein (EGFP), mCherry and cyan fluorescent protein (CFP) were selected by fluorescence-activated cell sorting (FACS) (Fig. 1d)

Summary

Introduction

Dengue virus (DENV) is spread from human to human through the bite of the female Aedes aegypti mosquito and leads to about 100 million clinical infections yearly. Defective interfering particles (DIPs) are considered a promising antiviral approach but infectious virus contamination has limited their development. A DENV-derived DIP production cell line was developed that continuously produced DENV-free DIPs. The DIPs contained and could deliver to cells a DENV serotype 2 subgenomic defective-interfering RNA, which was originally discovered in DENV infected patients. A hallmark of DIPs is that their production requires a WT helper virus that provides all of the necessary viral enzymes, cofactors and RNA packaging mechanisms so that DI RNA can replicate, be assembled into DIPs that will be secreted by infected cells[16]. DIPs parasitize cellular and viral resources required by WT viruses replication[13], and may stimulate cellular innate antiviral responses[22]. The use of DIPs as therapy or prophylaxis has been hampered by DIP preparations that are contaminated by infectious helper virus that is impractical to remove

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