A Beneficiary Role for Neuraminidase in Influenza Virus Penetration through the Respiratory Mucus

Yang, Xiaoyun; Steukers, Lennert; Forier, Katrien; Xiong, Ranhua; Braeckmans, Kevin; Reeth, Kristien Van; Nauwynck, Hans

doi:10.1371/journal.pone.0110026

Cited by 95 publications

(94 citation statements)

References 50 publications

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“…This is an important host defense mechanism, as pathogens trapped in mucus are expelled via mucociliary clearance before they are able to bind to cells and initiate an infection (27,29). The interaction between NA and mucins was hypothesized as early as 1943; more recently, by studying viral binding to MDCK cells, A549 cells, erythrocytes, and isolated respiratory mucus, NA activity has been shown to be important in initiating infection and viral penetration of mucus in vitro (18,20,(30)(31)(32). Mucus overlying frozen human trachea/bronchial tissue sections bound to viruses in a manner which appeared to be mediated by sialic acids (20).…”

Section: Discussionmentioning

confidence: 99%

Pandemic Swine H1N1 Influenza Viruses with Almost Undetectable Neuraminidase Activity Are Not Transmitted via Aerosols in Ferrets and Are Inhibited by Human Mucus but Not Swine Mucus

Zanin

Marathe

Wong

et al. 2015

J Virol

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A balance between the functions of the influenza virus surface proteins hemagglutinin (HA) and neuraminidase (NA) is thought to be important for the transmission of viruses between humans. Here we describe two pandemic H1N1 viruses, A/swine/Virginia/1814-1/2012 and A/swine/Virginia/1814-2/2012 (pH1N1 low -1 and -2, respectively), that were isolated from swine symptomatic for influenza. The enzymatic activity of the NA of these viruses was almost undetectable, while the HA binding affinity for ␣2,6 sialic acids was greater than that of the highly homologous pH1N1 viruses A/swine/Pennsylvania/2436/2012 and A/swine/Minnesota/2499/2012 (pH1N1-1 and -2), which exhibited better-balanced HA and NA activities. The in vitro growth kinetics of pH1N1 low and pH1N1 viruses were similar, but aerosol transmission of pH1N1 low -1 was abrogated and transmission via direct contact in ferrets was significantly impaired compared to pH1N1-1, which transmitted by direct and aerosol contact. In normal human bronchial epithelial cells, pH1N1 low -1 was significantly inhibited by mucus but pH1N1-1 was not. In Madin-Darby canine kidney cell cultures overlaid with human or swine mucus, human mucus inhibited pH1N1 low -1 but swine mucus did not. These data show that the interaction between viruses and mucus may be an important factor in viral transmissibility and could be a barrier for interspecies transmission between humans and swine for influenza viruses. IMPORTANCEA balance between the functions of the influenza virus surface proteins hemagglutinin (HA) and neuraminidase (NA) is thought to be important for transmission of viruses from swine to humans. Here we show that a swine virus with extremely functionally mismatched HA and NAs (pH1N1 low -1) cannot transmit via aerosol in ferrets, while another highly homologous virus with HA and NAs that are better matched functionally (pH1N1-1) can transmit via aerosol. These viruses show similar growth kinetics in Madin-Darby canine kidney (MDCK) cells, but pH1N1 low -1 is significantly inhibited by mucus in normal human bronchial epithelial cells whereas pH1N1-1 is not. Further, human mucus could inhibit these viruses, but swine mucus could not. These data show that the interaction between viruses and mucus may be an important factor in viral transmissibility and could be a species barrier between humans and swine for influenza viruses. H emagglutinin (HA) and neuraminidase (NA), the surface glycoproteins of influenza virus, play vital roles in the virus life cycle. HA binds to sialic acids on the cell surface, initiating fusion of the cell and viral membranes. NA enzymatically cleaves sialic acids from glycans on the host cell surface, facilitating the release of budding progeny viruses (1, 2). Thus, HA and NA have opposing roles in the viral life cycle and both are required for viral replication. A functional balance between HA and NA is important for both efficient replication and respiratory droplet transmission in humans. Four pandemic human viruses, A/California/04/2009 (H1N1) (...

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Section: Discussionmentioning

confidence: 99%

Pandemic Swine H1N1 Influenza Viruses with Almost Undetectable Neuraminidase Activity Are Not Transmitted via Aerosols in Ferrets and Are Inhibited by Human Mucus but Not Swine Mucus

Zanin

Marathe

Wong

et al. 2015

J Virol

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“…11 Others have shown that the influenza virus is immobilized in airway mucus through adhesive interactions of the virus with sialic acid on mucins found in the gel layer, as influenza requires the same specific binding to sialic acid on cell-surface glycans to mediate epithelial cell entry. 89 The influenza envelope contains a sialic-acid cleaving enzyme, neuraminidase, which was shown to facilitate its passage through the mucus gel layer. 89 Neutralizing antibodies against inhaled gene vectors can also impair penetration through the airway mucus gel layer.…”

Section: Trapping Mechanismsmentioning

confidence: 99%

“…89 The influenza envelope contains a sialic-acid cleaving enzyme, neuraminidase, which was shown to facilitate its passage through the mucus gel layer. 89 Neutralizing antibodies against inhaled gene vectors can also impair penetration through the airway mucus gel layer. Gene vectors coated with a sufficient number of antibodies are subject to multivalent adhesive trapping in mucus since the Fc region of antibodies binds, albeit with low affinity, to mucus.…”

Section: Trapping Mechanismsmentioning

confidence: 99%

The Mucus Barrier to Inhaled Gene Therapy

et al. 2016

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Recent evidence suggests that the airway mucus gel layer may be impermeable to the viral and synthetic gene vectors used in past inhaled gene therapy clinical trials for diseases like cystic fibrosis. These findings support the logic that inhaled gene vectors that are incapable of penetrating the mucus barrier are unlikely to provide meaningful benefit to patients. In this review, we discuss the biochemical and biophysical features of mucus that contribute its barrier function, and how these barrier properties may be reinforced in patients with lung disease. We next review biophysical techniques used to assess the potential ability of gene vectors to penetrate airway mucus. Finally, we provide new data suggesting that fresh human airway mucus should be used to test the penetration rates of gene vectors. The physiological barrier properties of spontaneously expectorated CF sputum remained intact up to 24 hours after collection when refrigerated at 4 °C. Conversely, the barrier properties were significantly altered after freezing and thawing of sputum samples. Gene vectors capable of overcoming the airway mucus barrier hold promise as a means to provide the widespread gene transfer throughout the airway epithelium required to achieve meaningful patient outcomes in inhaled gene therapy clinical trials.

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“…Viral purification was performed as described by Yang et al (2014). Briefly, culture fluids of EHV-1-infected (03P37 and 97P70) RK-13 cells were clarified and virus was collected after centrifugation.…”

Section: Cell Viabilitymentioning

confidence: 99%

Entry of equid herpesvirus 1 into CD172a+ monocytic cells

Laval

Favoreel

Cleemput

et al. 2016

Journal of General Virology

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Equid herpesvirus 1 (EHV-1) causes respiratory disease, abortion and neurological disorders in horses. Cells from the myeloid lineage (CD172a In contrast, we found that treatment of cells with neuraminidase (NA) reduced infection by 85-100 % compared with untreated cells, whilst NA treatment of virus had no effect on infection. This shows that sialic acid residues present on CD172a + cells are essential in the initiation of EHV-1 infection. We found that a V b 3 integrins are involved in the post-binding stage of CD172a + cell infection. Using pharmacological inhibitors, we showed that EHV-1 does not enter CD172a + cells via a clathrin-or caveolae-dependent endocytic pathway, nor by macropinocytosis, but requires cholesterol, tyrosine kinase, actin, dynamin and endosomal acidification, pointing towards a phagocytic mechanism. Overall, these results show that the narrow tropism of EHV-1 amongst CD172a + cells is determined by the presence of specific cellular receptors.

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A Beneficiary Role for Neuraminidase in Influenza Virus Penetration through the Respiratory Mucus

Cited by 95 publications

References 50 publications

Pandemic Swine H1N1 Influenza Viruses with Almost Undetectable Neuraminidase Activity Are Not Transmitted via Aerosols in Ferrets and Are Inhibited by Human Mucus but Not Swine Mucus

Pandemic Swine H1N1 Influenza Viruses with Almost Undetectable Neuraminidase Activity Are Not Transmitted via Aerosols in Ferrets and Are Inhibited by Human Mucus but Not Swine Mucus

The Mucus Barrier to Inhaled Gene Therapy

Entry of equid herpesvirus 1 into CD172a+ monocytic cells

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