Abstract
Influenza A virus (IAV) is a major human pathogen associated with significant morbidity and mortality worldwide. Through serial passage in mice, we generated a recombinant pdmH1N1 2009 IAV, A/Guangdong/GLW/2018 (GLW/18-MA), which encodes an mCherry gene fused to the C-terminal of a polymerase acidic (PA) segment and demonstrated comparable growth kinetics to the wild-type. Nine mutations were identified in the GLW/18-MA genome: PA (I61M, E351G, and G631S), NP (E292G), HA1 (T164I), HA2 (N117S and P160S), NA (W61R), and NEP (K44R). The recombinant IAV reporter expresses mCherry, a red fluorescent protein, at a high level and maintains its genetic integrity after five generations of serial passages in Madin-Darby Canine Kidney cells (MDCK) cells. Moreover, the imaging is noninvasive and permits the monitoring of infection in living mice. Treatment with oseltamivir or baicalin followed by infection with the reporter IAV led to a decrease in fluorescent protein signal in living mice. This result demonstrates that the IAV reporter virus is a powerful tool to study viral pathogenicity and transmission and to develop and evaluate novel anti-viral drugs, inhibitors, and vaccines in the future.
Highlights
Influenza A virus (IAV) is the most widespread virus worldwide, which causes respiratory diseases in avian and mammalian host species (Webster, 2002)
All eight Viral RNAs (vRNAs) are selectively packaged into each progeny virion through segment-specific genome-packaging signal sequences located in the noncoding and terminal coding regions of both the 5′ and the 3′ ends of the vRNAs (Goto et al, 2013)
Most of them were derived from the PR8 or WSN laboratory-adapted isolates, which may not be suitable for the study of contemporary epidemics (Kittel et al, 2004; Gao et al, 2010; Martinez-Sobrido et al, 2010; Pan et al, 2013; Fukuyama et al, 2015)
Summary
Influenza A virus (IAV) is the most widespread virus worldwide, which causes respiratory diseases in avian and mammalian host species (Webster, 2002). Outbreaks of avian influenza viruses (i.e., H5N1, H7N9, and H9N2) in humans, not resulting in a pandemic, show that exposure to infected poultry is the main risk factor for infection (Centers for Disease, C. and Prevention, 1997; Song and Qin, 2020; Song et al, 2021). A pdmH1N1 Reporter Virus subtypes, which may result in novel pandemic strains that threaten human health (Steel and Lowen, 2014). The latest development of in vivo bioluminescence or fluorescence imaging technology enables scientists to directly monitor cell activity in living organisms (Kim et al, 2015). The use of sensitive optical detection equipment allows researchers to directly analyze molecules, cells, and tissues from a range of living systems and investigate their real-time dynamics
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