Generation and Characterization of an Influenza D Reporter Virus.

Publisher: MDPI Country of Publication: Switzerland NLM ID: 101509722 Publication Model: Electronic Cited Medium: Internet ISSN: 1999-4915 (Electronic) Linking ISSN: 19994915 NLM ISO Abbreviation: Viruses Subsets: MEDLINE

Original Publication: Basel, Switzerland : MDPI

Influenza, Human*/genetics ; Thogotovirus*/genetics ; Orthomyxoviridae Infections* ; Orthomyxoviridae*/genetics; Cattle ; Animals ; Swine ; Humans ; Deltainfluenzavirus ; Viral Proteins/genetics ; Genes, Reporter ; Antiviral Agents/pharmacology

Influenza D virus (IDV) can infect various livestock animals, such as cattle, swine, and small ruminants, and was shown to have zoonotic potential. Therefore, it is important to identify viral factors involved in the broad host tropism and identify potential antiviral compounds that can inhibit IDV infection. Recombinant reporter viruses provide powerful tools for studying viral infections and antiviral drug discovery. Here we present the generation of a fluorescent reporter IDV using our previously established reverse genetic system for IDV. The mNeonGreen (mNG) fluorescent reporter gene was incorporated into the IDV non-structural gene segment as a fusion protein with the viral NS1 or NS2 proteins, or as a separate protein flanked by two autoproteolytic cleavage sites. We demonstrate that only recombinant reporter viruses expressing mNG as an additional separate protein or as an N-terminal fusion protein with NS1 could be rescued, albeit attenuated, compared to the parental reverse genetic clone. Serial passaging experiments demonstrated that the mNG gene is stably integrated for up to three passages, after which internal deletions accumulate. We conducted a proof-of-principle antiviral screening with the established fluorescent reporter viruses and identified two compounds influencing IDV infection. These results demonstrate that the newly established recombinant IDV reporter virus can be applied for antiviral drug discovery and monitoring viral replication, adding a new molecular tool for investigating IDV.

Chem Biol. 2013 Jul 25;20(7):912-21. (PMID: 23890009)
J Virol. 2020 Jan 31;94(4):. (PMID: 31776281)
Sci Rep. 2015 Jun 12;5:11346. (PMID: 26068081)
Nat Methods. 2012 Jun 28;9(7):676-82. (PMID: 22743772)
PLoS One. 2015 Mar 27;10(3):e0121491. (PMID: 25816132)
Viruses. 2021 Mar 18;13(3):. (PMID: 33803792)
Virology. 2015 Feb;476:206-216. (PMID: 25553516)
J Virol. 2020 May 4;94(10):. (PMID: 32102883)
Viruses. 2021 Apr 17;13(4):. (PMID: 33920517)
mBio. 2014 Mar 04;5(2):e00031-14. (PMID: 24595369)
Emerg Infect Dis. 2019 Oct;25(10):1969-1972. (PMID: 31287050)
Emerg Infect Dis. 2017 Aug;23(8):1392-1396. (PMID: 28726609)
Viruses. 2020 Sep 29;12(10):. (PMID: 33003329)
mBio. 2021 Dec 21;12(6):e0295921. (PMID: 34809454)
PLoS Pathog. 2013 Feb;9(2):e1003176. (PMID: 23408893)
Viruses. 2019 Jun 05;11(6):. (PMID: 31195597)
Sci Rep. 2016 Jun 06;6:27459. (PMID: 27264908)
Zoonoses Public Health. 2023 Mar;70(2):166-170. (PMID: 36370131)
J Clin Virol. 2016 Aug;81:31-3. (PMID: 27294672)
Viruses. 2019 Apr 24;11(4):. (PMID: 31022887)
Microorganisms. 2020 Nov 26;8(12):. (PMID: 33256227)
Viruses. 2015 Oct 13;7(10):5319-27. (PMID: 26473913)
Emerg Infect Dis. 2017 Sep;23(9):1556-1559. (PMID: 28820371)
J Virol. 2015 Dec;89(23):12226-31. (PMID: 26401044)
Virus Res. 2016 Feb 2;213:69-81. (PMID: 26590325)
PLoS One. 2016 Oct 5;11(10):e0163962. (PMID: 27706213)
Front Microbiol. 2019 Dec 17;10:2862. (PMID: 31921042)
Viruses. 2021 Oct 27;13(11):. (PMID: 34834971)
PLoS One. 2015 Dec 30;10(12):e0146021. (PMID: 26717316)
Front Immunol. 2022 Aug 19;13:970325. (PMID: 36059535)
Front Cell Infect Microbiol. 2022 Jan 20;11:827790. (PMID: 35127568)
J Virol. 2019 Oct 15;93(21):. (PMID: 31413133)
Vet Microbiol. 2015 Nov 18;180(3-4):281-285. (PMID: 26414999)
Blood. 2010 Apr 15;115(15):3109-17. (PMID: 20130243)
PLoS Negl Trop Dis. 2017 Apr 12;11(4):e0005540. (PMID: 28403145)
Proc Natl Acad Sci U S A. 2010 Jun 22;107(25):11531-6. (PMID: 20534532)
Nucleic Acids Res. 2012 Apr;40(7):3143-51. (PMID: 22140113)
J Microsc. 2013 Oct;252(1):89-91. (PMID: 23906423)
Nat Methods. 2013 May;10(5):407-9. (PMID: 23524392)

310030_179260 Switzerland SNSF_ Swiss National Science Foundation

Keywords: Influenza D Virus (IDV); antivirals; fluorescent reporter virus; reverse genetics

0 (Viral Proteins)
0 (Antiviral Agents)

Date Created: 20231223 Date Completed: 20231225 Latest Revision: 20240523

20240523

PMC10747006

10.3390/v15122444

38140686