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Influenza A penetrates host mucus by cleaving sialic acids with neuraminidase
Miriam Cohen1, Xing-Quan Zhang2, Hooman P Senaati1, Hui-Wen Chen23, Nissi M Varki1, Robert T Schooley2 and Pascal Gagneux1*
* Corresponding author: Pascal Gagneux pgagneux@ucsd.edu
Author Affiliations
1 Department of Cellular and Molecular Medicine, University of California San Diego, 9500 Gilman Dr, La Jolla 92093, California, USA
2 Division of Infectious Disease, University of California San Diego, 9500 Gilman Dr, La Jolla 92093, California, USA
3 Present address: School of Veterinary Medicine, National Taiwan University, 1 Sec. 4 Roosevelt Rd, Taipei 10617, Taiwan
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Virology Journal 2013, 10:321 doi:10.1186/1743-422X-10-321
The electronic version of this article is the complete one and can be found online at: http://www.virologyj.com/content/10/1/321
Received: 17 September 2013
Accepted: 14 October 2013
Published: 22 November 2013
? 2013 Cohen et al.; licensee BioMed Central Ltd.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
Abstract
Background
Influenza A virus (IAV) neuraminidase (NA) cleaves sialic acids (Sias) from glycans. Inhibiting NA with oseltamivir suppresses both viral infection, and viral release from cultured human airway epithelial cells. The role of NA in viral exit is well established: it releases budding virions by cleaving Sias from glycoconjugates on infected cells and progeny virions. The role of NA in viral entry remains unclear. Host respiratory epithelia secrete a mucus layer rich in heavily sialylated glycoproteins; these could inhibit viral entry by mimicking sialylated receptors on the cell surface. It has been suggested that NA allows influenza to penetrate the mucus by cleaving these sialylated decoys, but the exact mechanism is not yet established.
Methods
We tested IAV interaction with secreted mucus using frozen human trachea/bronchus tissue sections, and bead-bound purified human salivary mucins (HSM) and purified porcine submaxillary mucins (PSM). The protective effect of mucus was analyzed using MDCK cells coated with purified HSM and PSM with known Sia content. Oseltamivir was used to inhibit NA activity, and the fluorescent reporter substrate, 4MU-Neu5Ac, was used to quantify NA activity.
Results
IAV binds to the secreted mucus layer of frozen human trachea/bronchus tissues in a Sia dependent manner. HSM inhibition of IAV infection is Sia dose-dependent, but PSM cannot inhibit infection of underlying cells. HSM competitively inhibits NA cleavage of 4MU-Neu5Ac, reporter substrate. Human IAV effectively cleaves Sias from HSM but not from PSM, and binds to HSM but not to PSM.
Conclusion
IAV interacts with human mucus on frozen tissue sections and mucus-coated beads. Inhibition of IAV infection by sialylated human mucus is dose-dependent, and enhanced when NA is inhibited with oseltamivir. Thus NA cleaves sialylated decoys during initial stages of infection. Understanding IAV interactions with host mucins is a promising new avenue for drug development.
Keywords:
Influenza A; Sialic acids; Mucus; Neuraminidase; Infection; Saliva
full article
http://www.virologyj.com/content/10/1/321
Miriam Cohen1, Xing-Quan Zhang2, Hooman P Senaati1, Hui-Wen Chen23, Nissi M Varki1, Robert T Schooley2 and Pascal Gagneux1*
* Corresponding author: Pascal Gagneux pgagneux@ucsd.edu
Author Affiliations
1 Department of Cellular and Molecular Medicine, University of California San Diego, 9500 Gilman Dr, La Jolla 92093, California, USA
2 Division of Infectious Disease, University of California San Diego, 9500 Gilman Dr, La Jolla 92093, California, USA
3 Present address: School of Veterinary Medicine, National Taiwan University, 1 Sec. 4 Roosevelt Rd, Taipei 10617, Taiwan
For all author emails, please log on.
Virology Journal 2013, 10:321 doi:10.1186/1743-422X-10-321
The electronic version of this article is the complete one and can be found online at: http://www.virologyj.com/content/10/1/321
Received: 17 September 2013
Accepted: 14 October 2013
Published: 22 November 2013
? 2013 Cohen et al.; licensee BioMed Central Ltd.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
Abstract
Background
Influenza A virus (IAV) neuraminidase (NA) cleaves sialic acids (Sias) from glycans. Inhibiting NA with oseltamivir suppresses both viral infection, and viral release from cultured human airway epithelial cells. The role of NA in viral exit is well established: it releases budding virions by cleaving Sias from glycoconjugates on infected cells and progeny virions. The role of NA in viral entry remains unclear. Host respiratory epithelia secrete a mucus layer rich in heavily sialylated glycoproteins; these could inhibit viral entry by mimicking sialylated receptors on the cell surface. It has been suggested that NA allows influenza to penetrate the mucus by cleaving these sialylated decoys, but the exact mechanism is not yet established.
Methods
We tested IAV interaction with secreted mucus using frozen human trachea/bronchus tissue sections, and bead-bound purified human salivary mucins (HSM) and purified porcine submaxillary mucins (PSM). The protective effect of mucus was analyzed using MDCK cells coated with purified HSM and PSM with known Sia content. Oseltamivir was used to inhibit NA activity, and the fluorescent reporter substrate, 4MU-Neu5Ac, was used to quantify NA activity.
Results
IAV binds to the secreted mucus layer of frozen human trachea/bronchus tissues in a Sia dependent manner. HSM inhibition of IAV infection is Sia dose-dependent, but PSM cannot inhibit infection of underlying cells. HSM competitively inhibits NA cleavage of 4MU-Neu5Ac, reporter substrate. Human IAV effectively cleaves Sias from HSM but not from PSM, and binds to HSM but not to PSM.
Conclusion
IAV interacts with human mucus on frozen tissue sections and mucus-coated beads. Inhibition of IAV infection by sialylated human mucus is dose-dependent, and enhanced when NA is inhibited with oseltamivir. Thus NA cleaves sialylated decoys during initial stages of infection. Understanding IAV interactions with host mucins is a promising new avenue for drug development.
Keywords:
Influenza A; Sialic acids; Mucus; Neuraminidase; Infection; Saliva
full article
http://www.virologyj.com/content/10/1/321