Functional Characterization and Direct Comparison of Influenza A, B, C, and D NS1 Proteins in vitro and in vivo

Nogales, Aitor; Aydillo, Teresa; Ávila‐Pérez, Ginés; Escalera, Alba; Chiem, Kevin; Cadagan, Richard; DeDiego, Marta L.; Feng, Liang; Garcı́a-Sastre, Adolfo; Martínez-Sobrido, Luis

doi:10.3389/fmicb.2019.02862

Cited by 31 publications

(43 citation statements)

References 120 publications

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“…Indeed, NS1 has been described to positively regulate transcription/replication of the IAV genome and translation of viral mRNAs (for a review on NS1, see Hale et al, 2008). NS1 proteins of IAV and IBV share limited sequence homology, yet both have the ability to counteract the type 1 and type 3 IFN responses and are essential for efficient viral growth (Dauber et al, 2004(Dauber et al, , 2006Hai et al, 2008;Patzina et al, 2017;Nogales et al, 2019). Despite homologous functions, NS1 B has been shown to not fully complement the functions of NS1 A in the context of recombinant IAV viruses expressing heterotypic NS1, suggesting that other functions specific for NS1 A are not present in NS1 B (Nogales et al, 2019).…”

Section: Discussionmentioning

confidence: 99%

“…NS1 proteins of IAV and IBV share limited sequence homology, yet both have the ability to counteract the type 1 and type 3 IFN responses and are essential for efficient viral growth (Dauber et al, 2004(Dauber et al, , 2006Hai et al, 2008;Patzina et al, 2017;Nogales et al, 2019). Despite homologous functions, NS1 B has been shown to not fully complement the functions of NS1 A in the context of recombinant IAV viruses expressing heterotypic NS1, suggesting that other functions specific for NS1 A are not present in NS1 B (Nogales et al, 2019). One hypothesis is that, in the context of IAV-IBV co-infections, IAV-NS1 could exert effects similar to IAV RNAs on IBV RNAs and increase transcription/replication or translation of the IBV genome.…”

Section: Discussionmentioning

confidence: 99%

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Influenza B Virus Infection Is Enhanced Upon Heterotypic Co-infection With Influenza A Virus

et al. 2021

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Homotypic co-infections with influenza viruses are described to increase genetic population diversity, to drive viral evolution and to allow genetic complementation. Less is known about heterotypic co-infections between influenza A (IAV) and influenza B (IBV) viruses. Previous publications showed that IAV replication was suppressed upon co-infection with IBV. However, the effect of heterotypic co-infections on IBV replication was not investigated. To do so, we produced by reverse genetics a pair of replication-competent recombinant IAV (A/WSN/33) and IBV (B/Brisbane/60/2008) expressing a GFP and mCherry fluorescent reporter, respectively. A549 cells were infected simultaneously or 1 h apart at a high MOI with IAV-GFP or IBV-mCherry and the fluorescence was measured at 6 h post-infection by flow cytometry. Unexpectedly, we observed that IBV-mCherry infection was enhanced upon co-infection with IAV-GFP, and more strongly so when IAV was added 1 h prior to IBV. The same effect was observed with wild-type viruses and with various strains of IAV. Using UV-inactivated IAV or type-specific antiviral compounds, we showed that the enhancing effect of IAV infection on IBV infection was dependent on transcription/replication of the IAV genome. Our results, taken with available data in the literature, support the hypothesis that the presence of IAV proteins can enhance IBV genome expression and/or complement IBV defective particles.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Influenza B Virus Infection Is Enhanced Upon Heterotypic Co-infection With Influenza A Virus

et al. 2021

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“…Influenza A/Puerto Rico/08/1934 (PR8) H1N1 expressing mCherry and ∆NS1 viruses were previously described [ 38 , 69 ]. Viral titers (plaque forming units, PFU, per milliliter) were determined by standard plaque assay in MDCK cells [ 11 , 24 ]. MDCK cells expressing the green fluorescent protein (GFP) and firefly luciferase (FFluc) proteins under the control of the IFNβ promoter (MDCK pIFNβ-GFP/IFNβ-FFluc) were previously described [ 44 , 47 ].…”

Section: Methodsmentioning

confidence: 99%

“…Influenza A viruses (IAVs) are enveloped viruses containing a segmented genome of eight single-stranded RNA molecules of negative polarity that belong to the Orthomyxoviridae family [ 1 , 2 , 3 ]. Currently, there are four circulating influenza virus types: A, B, C, and D (IAV, IBV, ICV, and IDV, respectively), which are able to infect multiple mammalian (IAV, IBV, ICV and IDV) and avian (IAV) species [ 4 , 5 , 6 , 7 , 8 , 9 , 10 , 11 ]. IAVs are classified into different subtypes based on the viral surface hemagglutinin (HA; 18 subtypes) and neuraminidase (NA; 11 subtypes) glycoproteins.…”

Section: Introductionmentioning

confidence: 99%

“…Consequently, viruses lacking NS1 or containing truncated forms of NS1 are affected in viral replication in most cells and hosts, except for those that are deficient in IFN production and/or signaling [ 21 , 25 , 27 , 38 , 39 , 40 , 41 , 42 , 43 , 44 , 45 , 46 , 47 , 48 ]. IAV NS1 is able to modulate cell innate immune responses through different mechanisms that can be host- and viral strain-dependent [ 11 , 16 , 32 , 34 , 36 , 49 , 50 , 51 , 52 , 53 , 54 , 55 , 56 , 57 ]. However, despite multiple studies with IAV NS1, there are many gaps related to its role in pathogenesis, replication or the ability to cross the host species barrier.…”

Section: Introductionmentioning

confidence: 99%

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Replication-Competent ΔNS1 Influenza A Viruses Expressing Reporter Genes

Nogales

Schotsaert

Rathnasinghe

et al. 2021

Viruses

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The influenza A virus (IAV) is able to infect multiple mammalian and avian species, and in humans IAV is responsible for annual seasonal epidemics and occasional pandemics of respiratory disease with significant health and economic impacts. Studying IAV involves laborious secondary methodologies to identify infected cells. Therefore, to circumvent this requirement, in recent years, multiple replication-competent infectious IAV expressing traceable reporter genes have been developed. These IAVs have been very useful for in vitro and/or in vivo studies of viral replication, identification of neutralizing antibodies or antivirals, and in studies to evaluate vaccine efficacy, among others. In this report, we describe, for the first time, the generation and characterization of two replication-competent influenza A/Puerto Rico/8/1934 H1N1 (PR8) viruses where the viral non-structural protein 1 (NS1) was substituted by the monomeric (m)Cherry fluorescent or the NanoLuc luciferase (Nluc) proteins. The ΔNS1 mCherry was able to replicate in cultured cells and in Signal Transducer and Activator of Transcription 1 (STAT1) deficient mice, although at a lower extent than a wild-type (WT) PR8 virus expressing the same mCherry fluorescent protein (WT mCherry). Notably, expression of either reporter gene (mCherry or Nluc) was detected in infected cells by fluorescent microscopy or luciferase plate readers, respectively. ΔNS1 IAV expressing reporter genes provide a novel approach to better understand the biology and pathogenesis of IAV, and represent an excellent tool to develop new therapeutic approaches against IAV infections.

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Viral Fitness, Population Complexity, Host Interactions, and Resistance to Antiviral Agents

Domingo

García-Crespo

Soria

et al. 2023

Current Topics in Microbiology and Immunology

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Functional Characterization and Direct Comparison of Influenza A, B, C, and D NS1 Proteins in vitro and in vivo

Cited by 31 publications

References 120 publications

Influenza B Virus Infection Is Enhanced Upon Heterotypic Co-infection With Influenza A Virus

Influenza B Virus Infection Is Enhanced Upon Heterotypic Co-infection With Influenza A Virus

Replication-Competent ΔNS1 Influenza A Viruses Expressing Reporter Genes

Viral Fitness, Population Complexity, Host Interactions, and Resistance to Antiviral Agents

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