Investigating Influenza A Virus Infection: Tools To Track Infection and Limit Tropism

Fiege, Jessica K.; Langlois, Ryan A.

doi:10.1128/jvi.00462-15

Cited by 28 publications

(23 citation statements)

References 20 publications

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“…Recombinant viruses expressing fluorescent proteins have been generated to monitor viral replication, including DNA (e.g., Vaccinia virus (Diallo et al, 2011;Dominguez et al, 1998) and human cytomegalovirus (Marschall et al, 2000)), and double-stranded (e.g., HIV (Daelemans et al, 2005)), positive-stranded (e.g., murine hepatitis virus (Freeman et al, 2014)), negative-stranded segmented e.g., arenaviruses (Ortiz-Riano et al, 2013), and nonsegmented (e.g., Newcastle disease virus (Vigil et al, 2007) and Ebola virus (Towner et al, 2005)) RNA viruses. These recombinant systems are powerful tools to monitor viral infections in real time (Fiege and Langlois, 2015;Fukuyama et al, 2015;Manicassamy et al, 2010), to evaluate antivirals (Nogales et al, 2014a;Towner et al, 2005), and to identify NAb (Baker et al, 2015b;Nogales et al, 2014a;Rimmelzwaan et al, 2011).…”

Section: Discussionmentioning

confidence: 99%

“…With the development of reverse genetics techniques, similar approaches have been implemented for the generation and characterization of replication-competent fluorescentexpressing IAVs (Avilov et al, 2012;Baker et al, 2015b;Fiege and Langlois, 2015;Fukuyama et al, 2015;Manicassamy et al, 2010;Nogales et al, 2015Nogales et al, , 2014aReuther et al, 2015;Rimmelzwaan et al, 2011). However, similar methods have not been used for the generation of recombinant fluorescent-expressing IBVs.…”

Section: Discussionmentioning

confidence: 99%

“…Plasmid-based reverse genetics to generate recombinant influenza viruses (Fodor et al, 1999;Neumann et al, 1999) has significantly contributed to a better understanding of the biology of these important human respiratory pathogens (Engelhardt, 2013;Jackson et al, 2011a), for the identification and characterization of antivirals (Baker et al, 2014;Ozawa and Kawaoka, 2011;Roberts et al, 2015), and for the development of alternative influenza vaccines (Baker et al, 2015a;Nogales et al, 2014a;Subbarao and Katz, 2004). Reverse genetics have allowed for the generation of replication-competent IAVs expressing reporter genes as novel powerful tools to track viral infections without the requirement of secondary methodologies to detect viral-infected cells (Eckert et al, 2014;Fiege and Langlois, 2015;Fukuyama et al, 2015;Kittel et al, 2004;Manicassamy et al, 2010;Nogales et al, 2014a;Pan et al, 2013;Perez et al, 2013;Reuther et al, 2015;Tran et al, 2013). In contrast, similar approaches have not been implemented to the same extend for IBV.…”

Section: Introductionmentioning

confidence: 99%

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Replication-competent fluorescent-expressing influenza B virus

et al. 2016

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Influenza B viruses (IBVs) cause annual outbreaks of respiratory illness in humans and are increasingly recognized as a major cause of influenza-associated morbidity and mortality. Studying influenza viruses requires the use of secondary methodologies to identify virus-infected cells. To this end, replication-competent influenza A viruses (IAVs) expressing easily traceable fluorescent proteins have been recently developed. In contrast, similar approaches for IBV are mostly lacking. In this report, we describe the generation and characterization of replication-competent influenza B/Brisbane/60/2008 viruses expressing fluorescent mCherry or GFP fused to the C-terminal of the viral non-structural 1 (NS1) protein. Fluorescent-expressing IBVs display similar growth kinetics and plaque phenotype to wild-type IBV, while fluorescent protein expression allows for the easy identification of virus-infected cells. Without the need of secondary approaches to monitor viral infection, fluorescent-expressing IBVs represent an ideal approach to study the biology of IBV and an excellent platform for the rapid identification and characterization of antiviral therapeutics or neutralizing antibodies using high-throughput screening approaches. Lastly, fluorescent-expressing IBVs can be combined with the recently described reporter-expressing IAVs for the identification of novel therapeutics to combat these two important human respiratory pathogens.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Replication-competent fluorescent-expressing influenza B virus

et al. 2016

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“…Similarly, fusing of miR‐192 target sequence into IAV genome could attenuate influenza pathogenicity in mice . Furthermore, the ability of miRNA‐based attenuation method to determine virus fitness through introducing various number and location of target sites is superiority over classical viral attenuation methods . Together, these evidences suggested that the engineering flu genome virus containing the target sites of deregulated miRNAs in a species‐specific manner could be employed as a high‐throughput strategy which is able to provide a new class of flu IAV vaccines.…”

Section: Microrna and Exosome‐based Vaccinementioning

confidence: 99%

Influenza vaccine: Where are we and where do we go?

Keshavarz

Mirzaei

Salemi

et al. 2018

Reviews in Medical Virology

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Summary The alarming rise of morbidity and mortality caused by influenza pandemics and epidemics has drawn attention worldwide since the last few decades. This life‐threatening problem necessitates the development of a safe and effective vaccine to protect against incoming pandemics. The currently available flu vaccines rely on inactivated viral particles, M2e‐based vaccine, live attenuated influenza vaccine (LAIV) and virus like particle (VLP). While inactivated vaccines can only induce systemic humoral responses, LAIV and VLP vaccines stimulate both humoral and cellular immune responses. Yet, these vaccines have limited protection against newly emerging viral strains. These strains, however, can be targeted by universal vaccines consisting of conserved viral proteins such as M2e and capable of inducing cross‐reactive immune response. The lack of viral genome in VLP and M2e‐based vaccines addresses safety concern associated with existing attenuated vaccines. With the emergence of new recombinant viral strains each year, additional effort towards developing improved universal vaccine is warranted. Besides various types of vaccines, microRNA and exosome‐based vaccines have been emerged as new types of influenza vaccines which are associated with new and effective properties. Hence, development of a new generation of vaccines could contribute to better treatment of influenza.

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“…Despite the importance of this virus, far less is known about IBV relative to influenza A virus (IAV). Research into IAV has been greatly enhanced by several replication-competent reporter viruses (5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17). These reporter viruses have been crucial for identifying host factors, understanding basic viral pathogenesis, screening for antiviral compounds, and characterizing broadly reactive antibodies (6)(7)(8)(9)(10)(12)(13)(14)(15)(16)(17)(18)(19).…”

mentioning

confidence: 99%

Replication-Competent Influenza B Reporter Viruses as Tools for Screening Antivirals and Antibodies

Fulton

Palese

Heaton

2015

J Virol

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cInfluenza B virus is a human pathogen responsible for significant health and economic burden. Research into this pathogen has been limited by the lack of reporter viruses. Here we describe the development of both a replication-competent fluorescent influenza B reporter virus and bioluminescent influenza B reporter virus. Furthermore, we demonstrate these reporter viruses can be used to quickly monitor viral growth and permit the rapid screening of antiviral compounds and neutralizing antibodies. Influenza B virus (IBV) is a common human pathogen that causes yearly epidemics and significant disease in the pediatric population (1-4). Despite the importance of this virus, far less is known about IBV relative to influenza A virus (IAV). Research into IAV has been greatly enhanced by several replication-competent reporter viruses (5-17). These reporter viruses have been crucial for identifying host factors, understanding basic viral pathogenesis, screening for antiviral compounds, and characterizing broadly reactive antibodies (6)(7)(8)(9)(10)(12)(13)(14)(15)(16)(17)(18)(19). Lack of viruses encoding reporters has delayed similar progress for IBV.Development of a fluorescent influenza B reporter virus. We have previously published a description of an IAV reporter virus that contains the Gaussia luciferase gene directly behind the PB2 open reading frame (ORF) in segment 1 of A/Puerto Rico/8/1934 virus (PR8) (7). A 2A proteolytic cleavage site inserted between the PB2 ORF and the Gaussia luciferase gene allows for the cotranslational separation of the two proteins (20).Utilizing a similar approach, an influenza virus codon-optimized ORF of mNeonGreen (mNeon), a gene encoding a bright monomeric fluorescent protein (21), was cloned behind each of the three polymerase segments of the B/Yamagata/16/ 1988 (Ya88) virus (Fig. 1A). The reporter segments were rescued individually utilizing standard protocols (22-24) to generate the PB1 mNeon, PB2 mNeon, and PA mNeon viruses. Madin-Darby canine kidney (MDCK) cells were infected at a multiplicity of infection (MOI) of 0.5 for 18 h with the recombinant wild-type (rWT) Ya88 or one of the three reporter viruses (with all experiments carried out at 33°C). Tosyl phenylalanyl chloromethyl ketone (TPCK) trypsin, which is required for spread of viral progeny, was excluded from the infection medium. The cells were imaged or subjected to flow cytometry after being labeled with Hoechst stain (Thermo Scientific) or LIVE/DEAD stain (Life Technologies), respectively. By epifluorescence microscopy, the PB1 mNeon and PB2 mNeon viruses were bright and easily observed by 18 h postinfection, but the PA mNeon virus was less detectable (Fig. 1B to E). Flow cytometry (BD LSRIIA) recapitulated the microscopy results (Fig. 1F to I). Quantification of the flow cytometry data with FlowJo indicated that both the PB1 mNeon and PB2 mNeon viruses yielded the brightest signals ( Fig. 1J and K), and the PB1 mNeon virus was chosen for further study.The PB1 mNeon virus can rapidly monitor the growth of IBV in vi...

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Investigating Influenza A Virus Infection: Tools To Track Infection and Limit Tropism

Cited by 28 publications

References 20 publications

Replication-competent fluorescent-expressing influenza B virus

Replication-competent fluorescent-expressing influenza B virus

Influenza vaccine: Where are we and where do we go?

Replication-Competent Influenza B Reporter Viruses as Tools for Screening Antivirals and Antibodies

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