Entry Pathways of Herpes Simplex Virus Type 1 into Human Keratinocytes Are Dynamin- and Cholesterol-Dependent

Rahn, Elena; Petermann, Philipp; Hsu, Mei-Ju; Rixon, Frazer J.; Knebel-Mörsdorf, Dagmar

doi:10.1371/journal.pone.0025464

Cited by 81 publications

(96 citation statements)

References 50 publications

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“…How these alternative pathways correlate with nectin-1 internalization has still to be shown. The dual internalization mode in murine keratinocytes is in line with our results for human keratinocytes, where HSV-1 uptake occurs by direct fusion with the plasma membrane and by endocytic vesicles, in a process which requires dynamin and is dependent on cholesterol (27).…”

Section: Discussionsupporting

confidence: 89%

“…Murine epidermal whole mounts from wt or nectin-1-deficient mice were fixed with 3.4% formaldehyde as described previously (27) and then stained overnight with mouse anti-ICP0 (monoclonal antibody [MAb] 11060) (28) diluted 1:60, followed by washing with PBS-0.2% Tween 20 for 4 h, overnight incubation with anti-mouse immunoglobulin G (AF488) (Life Technologies) and DAPI (4=,6-diamidino-2-phenylindole), and washing with PBS-0.2% Tween 20 for 4 h, all at room temperature. Murine epidermal cryosections from wt or nectin-1-deficient mice were embedded in Tissue Tek (Sakura) and frozen in liquid nitrogen; 10-m frozen cross sections were then cut using a CM3050 (Leica) cryomicrotome.…”

Section: Rna Preparation and Reverse Transcription-pcr (Rt-pcr)mentioning

confidence: 99%

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Entry Mechanisms of Herpes Simplex Virus 1 into Murine Epidermis: Involvement of Nectin-1 and Herpesvirus Entry Mediator as Cellular Receptors

Petermann

Thier

Rahn

et al. 2015

J Virol

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Skin keratinocytes represent a primary entry site for herpes simplex virus 1 (HSV-1) in vivo. The cellular proteins nectin-1 and herpesvirus entry mediator (HVEM) act as efficient receptors for both serotypes of HSV and are sufficient for disease development mediated by HSV-2 in mice. How HSV-1 enters skin and whether both nectin-1 and HVEM are involved are not known. We addressed the impact of nectin-1 during entry of HSV-1 into murine epidermis and investigated the putative contribution of HVEM. Using ex vivo infection of murine epidermis, we showed that HSV-1 entered the basal keratinocytes of the epidermis very efficiently. In nectin-1-deficient epidermis, entry was strongly reduced. Almost no entry was observed, however, in nectin-1-deficient keratinocytes grown in culture. This observation correlated with the presence of HVEM on the keratinocyte surface in epidermis and with the lack of HVEM expression in nectin-1-deficient primary keratinocytes. Our results suggest that nectin-1 is the primary receptor in epidermis, while HVEM has a more limited role. For primary murine keratinocytes, on which nectin-1 acts as a single receptor, electron microscopy suggested that HSV-1 can enter both by direct fusion with the plasma membrane and via endocytic vesicles. Thus, we concluded that nectin-1 directs internalization into keratinocytes via alternative pathways. In summary, HSV-1 entry into epidermis was shown to strongly depend on the presence of nectin-1, but the restricted presence of HVEM can potentially replace nectin-1 as a receptor, illustrating the flexibility employed by HSV-1 to efficiently invade tissue in vivo. H erpes simplex viruses (HSV) are ubiquitous human pathogens which can cause a range of diseases, from mild, uncomplicated mucocutaneous lesions to life-threatening infections. HSV-1 is dominantly associated with orofacial infections and encephalitis, whereas HSV-2 more likely causes genital infections. To enter its human host, HSV must come into contact with mucosal surfaces, skin, or the cornea. During initial exposure on mucosa or skin, HSV targets epidermal keratinocytes and establishes a primary infection in the epithelium. Cellular entry of HSV relies on the interaction of several viral glycoproteins with various cell surface receptors (1, 2). The envelope glycoprotein D (gD) is essential for the entry process, and only after gD binding to a receptor is fusion with a cellular membrane induced (3). The major gD receptors mediating entry into mouse and human cells are herpesvirus entry mediator (HVEM) and nectin-1 (4-6). HVEM is a member of the tumor necrosis factor receptor superfamily which can activate either proinflammatory or inhibitory signaling pathways (7), while nectin-1 is an immunoglobulin-like cell adhesion molecule (8). A further gD receptor is 3-O-sulfated heparan sulfate, which may also play a role in HSV-1 entry into various cell types (9, 10).

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Section: Discussionsupporting

confidence: 89%

Section: Rna Preparation and Reverse Transcription-pcr (Rt-pcr)mentioning

confidence: 99%

Entry Mechanisms of Herpes Simplex Virus 1 into Murine Epidermis: Involvement of Nectin-1 and Herpesvirus Entry Mediator as Cellular Receptors

Petermann

Thier

Rahn

et al. 2015

J Virol

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“…Previously, we have shown that uptake into keratinocytes, the major cell type of the epidermis, depends on the multidomain GTPase, dynamin (37). To further characterize the internalization pathway in primary fibroblasts, we investigated the role of dynamin.…”

Section: Resultsmentioning

confidence: 99%

“…We have also previously shown that cholesterol is required for HSV-1 uptake into human primary keratinocytes, highlighting the role of cholesterol-rich lipid rafts (37). To investigate the role of cholesterol in primary dermal fibroblasts, we treated them with M␤CD, which depletes cholesterol from the plasma membrane.…”

Section: Resultsmentioning

confidence: 99%

Role of Nectin-1 and Herpesvirus Entry Mediator as Cellular Receptors for Herpes Simplex Virus 1 on Primary Murine Dermal Fibroblasts

Petermann

Rahn

Thier

et al. 2015

J Virol

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The cellular proteins nectin-1 and herpesvirus entry mediator (HVEM) can both mediate the entry of herpes simplex virus 1 (HSV-1). We have recently shown how these receptors contribute to infection of skin by investigating HSV-1 entry into murine epidermis. Ex vivo infection studies reveal nectin-1 as the primary receptor in epidermis, whereas HVEM has a more limited role. Although the epidermis represents the outermost layer of skin, the contribution of nectin-1 and HVEM in the underlying dermis is still open. Here, we analyzed the role of each receptor during HSV-1 entry in murine dermal fibroblasts that were deficient in expression of either nectin-1 or HVEM or both receptors. Because infection was not prevented by the absence of either nectin-1 or HVEM, we conclude that they can act as alternative receptors. Although HVEM was found to be highly expressed on fibroblasts, entry was delayed in nectin-1-deficient cells, suggesting that nectin-1 acts as the more efficient receptor. In the absence of both receptors, entry was strongly delayed leading to a much reduced viral spread and virus production. These results suggest an unidentified cellular component that acts as alternate but inefficient receptor for HSV-1 on dermal fibroblasts. Characterization of the cellular entry mechanism suggests that HSV-1 can enter dermal fibroblasts both by direct fusion with the plasma membrane and via endocytic vesicles and that this is not dependent on the presence or absence of nectin-1. Entry was also shown to require dynamin and cholesterol, suggesting comparable entry pathways in keratinocytes and dermal fibroblasts. IMPORTANCEHerpes simplex virus (HSV) is a human pathogen which infects its host via mucosal surfaces or abraded skin. To understand how HSV-1 overcomes the protective barrier of mucosa or skin and reaches its receptors in tissue, it is essential to know which receptors contribute to the entry into individual skin cells. Previously, we have explored the contribution of nectin-1 and herpesvirus entry mediator (HVEM) as receptors for HSV-1 entry into murine epidermis, where keratinocytes form the major cell type. Since the underlying dermis consists primarily of fibroblasts, we have now extended our study of HSV-1 entry to dermal fibroblasts isolated from nectin-1-or HVEM-deficient mice or from mice deficient in both receptors. Our results demonstrate a role for both nectin-1 and HVEM as receptors and suggest a further receptor which appears much less efficient.T o initiate infection, herpes simplex virus 1 (HSV-1) enters its human host via mucosal surfaces or abraded skin. HSV-1 entry into individual cells involves the interaction of several viral glycoproteins with various cell surface receptors (1, 2). The first step during entry is the attachment of virions to glycosaminoglycans, which facilitates the interaction with cellular receptors, leading to the fusion of the viral envelope with a cellular membrane. Fusion can either occur with the plasma membrane or with vesicle membranes after virions are int...

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“…One other example is herpes simplex virus type 1 (HSV-1). Although previous studies had indicated that the endocytosis of HSV-1 involves macropinocytosis [77], but Clement and Rahn have discovered that, the HSV-1, can take another route; a phagocytosis-like uptake that is facilitated by nectin-1, dynamin, and cholesterol-dependent manner [78,79]. In�uenza virus as mentioned above, enter host cells mainly via clathrin-mediated endocytosis.…”

Section: Unusual Endocytic Pathwaysmentioning

confidence: 99%

Molecular Signaling and Cellular Pathways for Virus Entry

Chi

Liu

2013

ISRN Virology

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The cell signaling plays a pivotal role in regulating cellular processes and is often manipulated by viruses as they rely on the functions offered by cells for their propagation. The first stage of their host life is to pass the genetic materials into the cell. Although some viruses can directly penetrate into cytosol, in fact, most virus entry into their host cells is through endocytosis. This machinery initiates with cell type specific cellular signaling pathways, and the signaling compounds can be proteins, lipids, and carbohydrates. The activation can be triggered in a very short time after virus binds on target cells, such as receptors. The signaling pathways involved in regulation of viral entry are wide diversity that often cross-talk between different endocytosis results. Furthermore, some viruses have the ability to use the multiple internalization pathways which leads to the regulation being even more complex. In this paper, we discuss some recent advances in our understanding of cellular pathways for virus entry, molecular signaling during virus entry, formation of endocytic vesicles, and the traffic.

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Entry Pathways of Herpes Simplex Virus Type 1 into Human Keratinocytes Are Dynamin- and Cholesterol-Dependent

Cited by 81 publications

References 50 publications

Entry Mechanisms of Herpes Simplex Virus 1 into Murine Epidermis: Involvement of Nectin-1 and Herpesvirus Entry Mediator as Cellular Receptors

Entry Mechanisms of Herpes Simplex Virus 1 into Murine Epidermis: Involvement of Nectin-1 and Herpesvirus Entry Mediator as Cellular Receptors

Role of Nectin-1 and Herpesvirus Entry Mediator as Cellular Receptors for Herpes Simplex Virus 1 on Primary Murine Dermal Fibroblasts

Molecular Signaling and Cellular Pathways for Virus Entry

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