A Defect in Influenza A Virus Particle Assembly Specific to Primary Human Macrophages

ABSTRACT Influenza A virus (IAV) propagates efficiently in epithelial cells, its primary target in the respiratory tract. In contrast, productive infection of most IAV strains is either blocked or highly inefficient in macrophages. The exact nature of the defect in IAV replication in human macrophag...

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Autores principales:	Sukhmani Bedi, Takeshi Noda, Yoshihiro Kawaoka, Akira Ono
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2018
Materias:	actin influenza macrophages plasma membrane virus assembly Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/d92cd2ca561a481d92ba0fcb467c797d
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spelling	oai:doaj.org-article:d92cd2ca561a481d92ba0fcb467c797d2021-11-15T15:58:21ZA Defect in Influenza A Virus Particle Assembly Specific to Primary Human Macrophages10.1128/mBio.01916-182150-7511https://doaj.org/article/d92cd2ca561a481d92ba0fcb467c797d2018-11-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.01916-18https://doaj.org/toc/2150-7511ABSTRACT Influenza A virus (IAV) propagates efficiently in epithelial cells, its primary target in the respiratory tract. In contrast, productive infection of most IAV strains is either blocked or highly inefficient in macrophages. The exact nature of the defect in IAV replication in human macrophages remains unknown. In this study, we showed that even compared to a monocytic cell line differentiated to macrophage-like cells, primary human monocyte-derived macrophages (MDM) are inefficient in IAV production, despite comparable levels of expression of viral glycoproteins at the plasma membrane. Correlative fluorescence scanning electron microscopy revealed that formation of budding structures at the cell surface is inefficient in MDM even though clustering of a viral glycoprotein, hemagglutinin (HA), is observed, suggesting that a step in IAV particle assembly is blocked in MDM. Using an in situ proximity ligation assay, we further determined that HA associates with neuraminidase (NA) but fails to associate with another viral transmembrane protein, M2, at the MDM plasma membrane. Notably, the defects in HA-M2 association and particle assembly in MDM were reversed upon cytochalasin D treatment that inhibits actin polymerization. These results suggest that HA-M2 association on the plasma membrane is a discrete step in IAV production, which is susceptible to suppression by actin cytoskeleton in MDM. Virus release remained inefficient in MDM upon cytochalasin D treatment, suggesting the presence of an additional defect(s) in virus release in this cell type. Overall, our study revealed the presence of multiple cell-type-specific mechanisms negatively regulating IAV production at the plasma membrane in MDM. IMPORTANCE Identification of host cell determinants promoting or suppressing replication of viruses has been aided by analyses of host cells that impose inherent blocks on viral replication. In this study, we show that primary human MDM, which are not permissive to IAV replication, fail to support virus particle formation. This defect is specific to primary human macrophages, since a human monocytic cell line differentiated to macrophage-like cells supports IAV particle formation. We further identified association between two viral transmembrane proteins, HA and M2, on the cell surface as a discrete assembly step, which is defective in MDM. Defective HA-M2 association and particle budding, but not virus release, in MDM are rescued by disruption of actin cytoskeleton, revealing a previously unknown, negative role for actin, which specifically targets an early step in the multistep IAV production. Overall, our study uncovered a host-mediated restriction of association between viral transmembrane components during IAV assembly.Sukhmani BediTakeshi NodaYoshihiro KawaokaAkira OnoAmerican Society for Microbiologyarticleactininfluenzamacrophagesplasma membranevirus assemblyMicrobiologyQR1-502ENmBio, Vol 9, Iss 5 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	actin influenza macrophages plasma membrane virus assembly Microbiology QR1-502
spellingShingle	actin influenza macrophages plasma membrane virus assembly Microbiology QR1-502 Sukhmani Bedi Takeshi Noda Yoshihiro Kawaoka Akira Ono A Defect in Influenza A Virus Particle Assembly Specific to Primary Human Macrophages
description	ABSTRACT Influenza A virus (IAV) propagates efficiently in epithelial cells, its primary target in the respiratory tract. In contrast, productive infection of most IAV strains is either blocked or highly inefficient in macrophages. The exact nature of the defect in IAV replication in human macrophages remains unknown. In this study, we showed that even compared to a monocytic cell line differentiated to macrophage-like cells, primary human monocyte-derived macrophages (MDM) are inefficient in IAV production, despite comparable levels of expression of viral glycoproteins at the plasma membrane. Correlative fluorescence scanning electron microscopy revealed that formation of budding structures at the cell surface is inefficient in MDM even though clustering of a viral glycoprotein, hemagglutinin (HA), is observed, suggesting that a step in IAV particle assembly is blocked in MDM. Using an in situ proximity ligation assay, we further determined that HA associates with neuraminidase (NA) but fails to associate with another viral transmembrane protein, M2, at the MDM plasma membrane. Notably, the defects in HA-M2 association and particle assembly in MDM were reversed upon cytochalasin D treatment that inhibits actin polymerization. These results suggest that HA-M2 association on the plasma membrane is a discrete step in IAV production, which is susceptible to suppression by actin cytoskeleton in MDM. Virus release remained inefficient in MDM upon cytochalasin D treatment, suggesting the presence of an additional defect(s) in virus release in this cell type. Overall, our study revealed the presence of multiple cell-type-specific mechanisms negatively regulating IAV production at the plasma membrane in MDM. IMPORTANCE Identification of host cell determinants promoting or suppressing replication of viruses has been aided by analyses of host cells that impose inherent blocks on viral replication. In this study, we show that primary human MDM, which are not permissive to IAV replication, fail to support virus particle formation. This defect is specific to primary human macrophages, since a human monocytic cell line differentiated to macrophage-like cells supports IAV particle formation. We further identified association between two viral transmembrane proteins, HA and M2, on the cell surface as a discrete assembly step, which is defective in MDM. Defective HA-M2 association and particle budding, but not virus release, in MDM are rescued by disruption of actin cytoskeleton, revealing a previously unknown, negative role for actin, which specifically targets an early step in the multistep IAV production. Overall, our study uncovered a host-mediated restriction of association between viral transmembrane components during IAV assembly.
format	article
author	Sukhmani Bedi Takeshi Noda Yoshihiro Kawaoka Akira Ono
author_facet	Sukhmani Bedi Takeshi Noda Yoshihiro Kawaoka Akira Ono
author_sort	Sukhmani Bedi
title	A Defect in Influenza A Virus Particle Assembly Specific to Primary Human Macrophages
title_short	A Defect in Influenza A Virus Particle Assembly Specific to Primary Human Macrophages
title_full	A Defect in Influenza A Virus Particle Assembly Specific to Primary Human Macrophages
title_fullStr	A Defect in Influenza A Virus Particle Assembly Specific to Primary Human Macrophages
title_full_unstemmed	A Defect in Influenza A Virus Particle Assembly Specific to Primary Human Macrophages
title_sort	defect in influenza a virus particle assembly specific to primary human macrophages
publisher	American Society for Microbiology
publishDate	2018
url	https://doaj.org/article/d92cd2ca561a481d92ba0fcb467c797d
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A Defect in Influenza A Virus Particle Assembly Specific to Primary Human Macrophages

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