Sustained replication of synthetic canine distemper virus defective genomes in vitro and in vivo

Natasha L. Tilston-Lunel, Stephen R. Welch, Sham Nambulli, Rory D. De Vries, Gregory W. Ho, David E. Wentworth, Reed Shabman, Stuart T. Nichol, Christina F. Spiropoulou, Rik L. De Swart, Linda J. Rennick, W. Paul Duprex*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

2 Citations (Scopus)

Abstract

Defective interfering (DI) genomes restrict viral replication and induce type I interferon. Since DI genomes have been proposed as vaccine adjuvants or therapeutic antiviral agents, it is important to understand their generation, delineate their mechanism of action, develop robust production capacities, assess their safety and in vivo longevity, and determine their long-term effects. To address this, we generated a recombinant canine distemper virus (rCDV) from an entirely synthetic molecular clone designed using the genomic sequence from a clinical isolate obtained from a free-ranging raccoon with distemper. rCDV was serially passaged in vitro to identify DI genomes that naturally arise during rCDV replication. Defective genomes were identified by Sanger and next-generation sequencing techniques, and predominant genomes were synthetically generated and cloned into T7-driven plasmids. Fully encapsidated DI particles (DIPs) were then generated using a rationally attenuated rCDV as a producer virus to drive DI genome replication. We demonstrate that these DIPs interfere with rCDV replication in a dose-dependent manner in vitro. Finally, we show sustained replication of a fluorescent DIP in experimentally infected ferrets over a period of 14 days. Most importantly, DIPs were isolated from the lymphoid tissues, which are a major site of CDV replication. Our established pipeline for detection, generation, and assaying DIPs is transferable to highly pathogenic paramyxoviruses and will allow qualitative and quantitative assessment of the therapeutic effects of DIP administration on disease outcome.

Original languageEnglish
Article numbere00537-21
JournalmSphere
Volume6
Issue number5
Early online date22 Sep 2021
DOIs
Publication statusPublished - Oct 2021

Bibliographical note

Funding Information:
This work was funded by the Defense Advanced Research Projects Agency (DARPA) INTERfering and Co-Evolving Prevention and Therapy (INTERCEPT) program (grant HR0011940493), Boston University, and the University of Pittsburgh.

Publisher Copyright:
© 2021 American Society for Microbiology. All rights reserved.

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