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iScience . A human H5N1 influenza virus expressing bioluminescence for evaluating viral infection and identifying therapeutic interventions

tetano

Editor, Senior Moderator
iScience


. 2025 Aug 21;28(9):113402.
doi: 10.1016/j.isci.2025.113402. eCollection 2025 Sep 19. A human H5N1 influenza virus expressing bioluminescence for evaluating viral infection and identifying therapeutic interventions

Ramya S Barre[SUP] 1 2 [/SUP], Ruby A Escobedo[SUP] 1 2 [/SUP], Esteban M Castro[SUP] 1 2 [/SUP], Michal Gazi[SUP] 1 [/SUP], Joshua D Castro[SUP] 1 [/SUP], Anastasija Cupic[SUP] 3 4 [/SUP], Mahmoud Bayoumi[SUP] 1 5 [/SUP], Nathaniel Jackson[SUP] 1 2 [/SUP], Chengin Ye[SUP] 1 [/SUP], Aitor Nogales[SUP] 6 [/SUP], Roy N Platt[SUP] 1 [/SUP], Ricardo Carrion Jr[SUP] 1 [/SUP], Timothy J C Anderson[SUP] 1 [/SUP], Adolfo García-Sastre[SUP] 3 7 8 9 10 11 [/SUP], Ahmed Mostafa[SUP] 1 12 [/SUP], Luis Martinez-Sobrido[SUP] 1 [/SUP]



Affiliations
Abstract

A multistate outbreak of highly pathogenic avian influenza virus (HPAIV) H5N1 in the United States dairy cattle was first reported on March 2024, followed by a zoonotic cattle-to-human virus transmission to a dairy farm worker in Texas. To facilitate real-time tracking of HPAIV H5N1, we generated a recombinant nanoluciferase (Nluc)-expressing H5N1 virus, HPhTX NSs-Nluc, by introducing an Nluc reporter into the non-structural gene of human A/Texas/37/2024 H5N1 (HPhTX). HPhTX NSs-Nluc replicated with kinetics and plaque morphology comparable to wild-type virus in vitro. In vivo and ex vivo, HPhTX NSs-Nluc allowed tracking viral infection in the living animals and their necropsied organs using in vivo imaging systems (IVISs). Treatment with baloxavir effectively inhibited HPhTX NSs-Nluc replication, comparable to wild-type virus, validating its applicability for high-throughput screening of potential antiviral therapeutics. These results demonstrate that HPhTX NSs-Nluc is a robust tool for studying H5N1 pathogenesis and assessing antiviral efficacy against HPAIV H5N1.

Keywords: Bioengineering; Methodology in biological sciences; Optical imaging; Virology.

 
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