A mutation in the receptor binding site enhances infectivity of 2009 H1N1 influenza hemagglutinin pseudotypes without changing antigenicity

Wang, Wei; Castelán-Vega, Juan Arturo; Jiménez-Alberto, Alicia; Vassell, Russell; Ye, Zhiping; Weiss, Carol D.

doi:10.1016/j.virol.2010.08.027

Cited by 20 publications

(32 citation statements)

References 39 publications

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“…It is of considerable importance to detect new antigenic changes occurring in HA protein when updating vaccine compositions. The results of our study are consistent with the results of others in terms of changes in number and location of glycosylation sites (20,21), mutations/conservation in important positions such as Q240R (22), V169T, and D239G (24,25), changes/conservation in active/binding sites (23,24), and conservation in important domains such as fusion peptide (24,25). Evaluation of changes in predicted ligandbinding sites, type and number of ligands was done in our study, which indicated a significant change in ligand binding site of 2009 isolates.…”

Section: Discussionsupporting

confidence: 93%

“…Addition of glycans to the HA is an important mechanism contributing to antigenic drift and therefore sustained circulation of influenza A virus in the human population (20,21). The (Table 3), in which some changes (creation/ deletion of glycosyla- Position 240, which is located in receptor binding site, is one of the important positions in HA (22). This position is conserved in sequences of 2006 to 2008 isolates, yet, from 2009 onwards, Q240R mutation has occurred in some isolates.…”

Section: Resultsmentioning

confidence: 99%

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A Bioinformatics Study of Complete Amino Acid Sequences’ Changes of Hemagglutinin Antigen of H1N1 Influenza Viruses in GenBank From Year 2006 to 2013 in Iran

Farhangi

Goliaei

Kavousi

et al. 2017

Jundishapur J Microbiol

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Section: Discussionsupporting

confidence: 93%

Section: Resultsmentioning

confidence: 99%

A Bioinformatics Study of Complete Amino Acid Sequences’ Changes of Hemagglutinin Antigen of H1N1 Influenza Viruses in GenBank From Year 2006 to 2013 in Iran

Farhangi

Goliaei

Kavousi

et al. 2017

Jundishapur J Microbiol

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“…As previously described (22), the full-length Q223R HA open reading frame (ORF) from A/Mexico/4108/2009 (MX HA) (GenBank accession no. GQ223112), which was previously reported to enhance pseudovirus infectivity (18), and the wild-type NA ORF from A/California/04/2009 (GenBank accession no. FJ966084) were amplified from the respective viruses by reverse transcription-PCR (RT-PCR) and placed into the CMV/R 8B expression plasmid, encoding HA from A/PR/8/34 (PR HA) (GenBank accession no.…”

Section: Methodsmentioning

confidence: 99%

“…We previously showed that the M2 protein enhanced (H1N1) pdm09 HA-pseudovirus infectivity when M2 was coexpressed with HA during pseudovirus production (18). However, the mechanism responsible for this effect remained unknown.…”

mentioning

confidence: 99%

Influenza Virus M2 Protein Ion Channel Activity Helps To Maintain Pandemic 2009 H1N1 Virus Hemagglutinin Fusion Competence during Transport to the Cell Surface

Alvarado-Facundo

Gao

Ribas‐Aparicio

et al. 2015

J Virol

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The influenza virus hemagglutinin (HA) envelope protein mediates virus entry by first binding to cell surface receptors and then fusing viral and endosomal membranes during endocytosis. Cleavage of the HA precursor (HA0) into a surface receptor-binding subunit (HA1) and a fusion-inducing transmembrane subunit (HA2) by host cell enzymes primes HA for fusion competence by repositioning the fusion peptide to the newly created N terminus of HA2. We previously reported that the influenza virus M2 protein enhances pandemic 2009 influenza A virus [(H1N1)pdm09] HA-pseudovirus infectivity, but the mechanism was unclear. In this study, using cell-cell fusion and HA-pseudovirus infectivity assays, we found that the ion channel function of M2 was required for enhancement of HA fusion and HA-pseudovirus infectivity. The M2 activity was needed only during HA biosynthesis, and proteolysis experiments indicated that M2 proton channel activity helped to protect (H1N1)pdm09 HA from premature conformational changes as it traversed low-pH compartments during transport to the cell surface. While M2 has previously been shown to protect avian influenza virus HA proteins of the H5 and H7 subtypes that have polybasic cleavage motifs, this study demonstrates that M2 can protect HA proteins from human H1N1 strains that lack a polybasic cleavage motif. This finding suggests that M2 proton channel activity may play a wider role in preserving HA fusion competence among a variety of HA subtypes, including HA proteins from emerging strains that may have reduced HA stability. IMPORTANCEInfluenza virus infects cells when the hemagglutinin (HA) surface protein undergoes irreversible pH-induced conformational changes after the virus is taken into the cell by endocytosis. HA fusion competence is primed when host cell enzymes cleave the HA precursor. The proton channel function of influenza virus M2 protein has previously been shown to protect avian influenza virus HA proteins that contain a polybasic cleavage site from pH-induced conformational changes during biosynthesis, but this effect is less well understood for human influenza virus HA proteins that lack polybasic cleavage sites. Using assays that focus on HA entry and fusion, we found that the M2 protein also protects (H1N1)pdm09 influenza A virus HA from premature conformational changes as it transits low-pH compartments during biosynthesis. This work suggests that M2 may play a wider role in preserving HA function in a variety of influenza virus subtypes that infect humans and may be especially important for HA proteins that are less stable. T he influenza virus hemagglutinin (HA) envelope protein mediates virus entry by binding to cell surface receptors, followed by endocytosis and fusion of the viral and endosomal membranes. Cellular proteases cleave the HA precursor (HA0) to generate the HA1 surface subunit, which mediates binding to cell surface sialic acid receptors, and the HA2 transmembrane subunit, which mediates membrane fusion between viral and endosomal membranes during e...

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“…An egg adaptation mutation leading to a Q223R substitution in HA was found to have occurred in the passaged viruses encoding both wild-type PB1 and PB1-V43I. The Q223R mutation in the receptor binding site improves infectivity in egg and in human cells without changing HA antigenicity (28,35). Interestingly, the K154N and G155E antigenic mutations were introduced into the HA of the PB1 wild-type vaccine virus following passages; however, these mutations were not found in the PB1-V43I vaccine strain (Tables 3 and 4).…”

Section: Discussionmentioning

confidence: 99%

Generation of a Genetically Stable High-Fidelity Influenza Vaccine Strain

Naito

Mori

Ushirogawa

et al. 2017

J Virol

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Vaccination is considered the most effective preventive means for influenza control. The development of a master virus with high growth and genetic stability, which may be used for the preparation of vaccine viruses by gene reassortment, is crucial for the enhancement of vaccine performance and efficiency of production. Here, we describe the generation of a high-fidelity and high-growth influenza vaccine master virus strain with a single V43I amino acid change in the PB1 polymerase of the high-growth A/Puerto Rico/8/1934 (PR8) master virus. The PB1-V43I mutation was introduced to increase replication fidelity in order to design an H1N1 vaccine strain with a low error rate. The PR8-PB1-V43I virus exhibited good replication compared with that of the parent PR8 virus. In order to compare the efficiency of egg adaptation and the occurrence of gene mutations leading to antigenic alterations, we constructed 6:2 genetic reassortant viruses between the A(H1N1)pdm09 and the PR8-PB1-V43I viruses; hemagglutinin (HA) and neuraminidase (NA) were from the A(H1N1)pdm09 virus, and the other genes were from the PR8 virus. Mutations responsible for egg adaptation mutations occurred in the HA of the PB1-V43I reassortant virus during serial egg passages; however, in contrast, antigenic mutations were introduced into the HA gene of the 6:2 reassortant virus possessing the wild-type PB1. This study shows that the mutant PR8 virus possessing the PB1 polymerase with the V43I substitution may be utilized as a master virus for the generation of high-growth vaccine viruses with high polymerase fidelity, low error rates of gene replication, and reduced antigenic diversity during virus propagation in eggs for vaccine production.IMPORTANCE Vaccination represents the most effective prophylactic option against influenza. The threat of emergence of influenza pandemics necessitates the ability to generate vaccine viruses rapidly. However, as the influenza virus exhibits a high mutation rate, vaccines must be updated to ensure a good match of the HA and NA antigens between the vaccine and the circulating strain. Here, we generated a genetically stable master virus of the A/Puerto Rico/8/1934 (H1N1) backbone encoding an engineered high-fidelity viral polymerase. Importantly, following the application of the high-fidelity PR8 backbone, no mutation resulting in antigenic change was introduced into the HA gene during propagation of the A(H1N1)pdm09 candidate vaccine virus. The low error rate of the present vaccine virus should decrease the risk of generating mutant viruses with increased virulence. Therefore, our findings are expected to be useful for the development of prepandemic vaccines and live attenuated vaccines with higher safety than that of the present candidate vaccines.

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A mutation in the receptor binding site enhances infectivity of 2009 H1N1 influenza hemagglutinin pseudotypes without changing antigenicity

Cited by 20 publications

References 39 publications

A Bioinformatics Study of Complete Amino Acid Sequences’ Changes of Hemagglutinin Antigen of H1N1 Influenza Viruses in GenBank From Year 2006 to 2013 in Iran

A Bioinformatics Study of Complete Amino Acid Sequences’ Changes of Hemagglutinin Antigen of H1N1 Influenza Viruses in GenBank From Year 2006 to 2013 in Iran

Influenza Virus M2 Protein Ion Channel Activity Helps To Maintain Pandemic 2009 H1N1 Virus Hemagglutinin Fusion Competence during Transport to the Cell Surface

Generation of a Genetically Stable High-Fidelity Influenza Vaccine Strain

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