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

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

doi:10.1128/jvi.01415-15

Cited by 34 publications

(48 citation statements)

References 44 publications

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“…Moreover, this is also the scenario in HeLa cells where there is 60-fold less nectin-1 mRNA present, suggesting that even low levels of nectin-1 can support HSV-1 entry. Together with our results on HeLa and nTERT cells presented here, nectin-1 has now been shown to be the preferred entry receptor over HVEM in a range of different cell types (37)(38)(39)(40)(41). As such, it could be argued that the relatively high level of nectin-1 expression in nTERT cells explains the rapid entry phenotype.…”

Section: Discussionsupporting

confidence: 81%

Herpes Simplex Virus 1 Enters Human Keratinocytes by a Nectin-1-Dependent, Rapid Plasma Membrane Fusion Pathway That Functions at Low Temperature

Sayers

Elliott

2016

J Virol

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Herpes simplex virus 1 (HSV-1) infects humans through stratified epithelia that are composed primarily of keratinocytes. The route of HSV-1 entry into keratinocytes has been the subject of limited investigation, but it is proposed to involve pH-dependent endocytosis, requiring the gD-binding receptor nectin-1. Here, we have utilized the nTERT human keratinocyte cell line as a new model for dissecting the mechanism of HSV-1 entry into the host. Although immortalized, these cells nonetheless retain normal growth and differentiation properties of primary cells. Using short interfering RNA (siRNA) depletion studies, we confirm that, despite nTERT cells expressing high levels of the alternative gD receptor HVEM, HSV-1 requires nectin-1, not HVEM, to enter these cells. Strikingly, virus entry into nTERT cells occurred with unusual rapidity, such that maximum penetration was achieved within 5 min. Moreover, HSV-1 was able to enter keratinocytes but not other cell types at temperatures as low as 7°C, conditions where endocytosis was shown to be completely inhibited. Transmission electron microscopy of early entry events at both 37°C and 7°C identified numerous examples of naked virus capsids located immediately beneath the plasma membrane, with no evidence of virions in cytoplasmic vesicles. Taken together, these results imply that HSV-1 uses the nectin-1 receptor to enter human keratinocyte cells via a previously uncharacterized rapid plasma membrane fusion pathway that functions at low temperature. These studies have important implications for current understanding of the relationship between HSV-1 and its relevant in vivo target cell. IMPORTANCEThe gold standard of antiviral treatment for any human virus infection is the prevention of virus entry into the host cell. In the case of HSV-1, primary infection in the human begins in the epidermis of the skin or the oral mucosa, where the virus infects keratinocytes, and it is therefore important to understand the molecular events involved in HSV-1 entry into this cell type. Nonetheless, few studies have looked specifically at entry into these relevant human cells. Our results reveal a new route for virus entry that is specific to keratinocytes, involves rapid entry, and functions at low temperatures. This may reflect the environmental conditions encountered by HSV-1 when entering its host through the skin and emphasizes the importance of studying virushost interactions in physiologically relevant cells. H erpes simplex virus type 1 (HSV-1) gains access to its human host through the epidermis at the oral mucosa, skin, or the cornea, where the majority of the cells are keratinocytes. Hence, although HSV-1 replication has been studied in many cell types in vitro, the role of the human keratinocyte in the HSV-1 life cycle makes it the most physiologically relevant cell type in which to unravel HSV-1 replication strategies important in its natural host. To date, there have been only a few studies on the mechanism of HSV-1 entry into human keratinocytes. While initial s...

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Section: Discussionsupporting

confidence: 81%

Herpes Simplex Virus 1 Enters Human Keratinocytes by a Nectin-1-Dependent, Rapid Plasma Membrane Fusion Pathway That Functions at Low Temperature

Sayers

Elliott

2016

J Virol

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“…Interestingly, these expression levels do not appear to correlate with their effectiveness as receptors. Despite its low level on fibroblasts, our results support nectin-1 as the major mediator of HSV-1 entry into both cell types of murine skin [3,4]. In the absence of nectin-1, HVEM can replace it as a receptor, and appears to do so more efficiently in fibroblasts than in keratinocytes.…”

supporting

confidence: 64%

“…Whether both internalization pathways lead to productive infection is difficult to determine although studies in human keratinocytes support endocytic uptake as contributing to HSV-1 entry [7]. In addition, we demonstrated that entry into skin cells is cholesterol-and dynamin-mediated [4,7]. Based on the known functions of dynamin, the finding that inhibition of dynamin GTPase activity results in a Editorial…”

Section: +mentioning

confidence: 83%

“…It seems clear that nectin-1 binding accounts primarily for the uptake mechanism, while HVEM binding may have a secondary effect of modulating the immune response by interfering with natural ligands. The rapid loss of nectin-1 from the surface of epidermal keratinocytes and dermal fibroblasts upon infection supports this assumption [3,4]. A further intriguing question is whether and how HSV-1 gains access to the cell-cell adhesion molecule nectin-1 in intact skin or mucosa where close cell-cell contacts might be expected to act as a barrier.…”

Section: +mentioning

confidence: 97%

“…The rapid loss of nectin-1 from the surface of epidermal keratinocytes and dermal fibroblasts upon infection supports this assumption [3,4]. A further intriguing question is whether and how HSV-1 gains access to the cell-cell adhesion molecule nectin-1 in intact skin or mucosa where close cell-cell contacts might be expected to act as a barrier.Characterization of the uptake pathway in murine skin suggests that HSV-1 enters into epidermal sheets, primary epidermal keratinocytes and primary dermal fibroblasts, both by direct fusion of the viral envelope with the plasma membrane and via endocytic vesicles [3,4]. Interestingly, this is not dependent on the presence or absence of nectin-1, suggesting that nectin-1 and HVEM can initiate both uptake modes.…”

mentioning

confidence: 97%

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2016

Oncotarget

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Herpes simplex virus (HSV) can cause a range of diseases in humans from mild uncomplicated mucocutaneous lesions to life-threatening infections. HSV 1 (HSV-1) is dominantly associated with orofacial infections and encephalitis, and can be devastating in immune-compromised hosts and newborns. Although entry into individual cells in culture and the immune response to HSV-1 have been studied in detail, we have little knowledge of how the virus invades actual target tissues or of which cellular factors determine susceptibility to infection in vivo. Two important factors are the cell surface receptors, herpesvirus entry mediator (HVEM) and nectin-1, which interact with the viral envelope glycoprotein D (gD) [1]. This interaction is essential for the fusion of the viral envelope with cellular membranes, allowing delivery of the nucleocapsid into the cytoplasm to initiate infection. Target tissues for HSV-1 in the human host are mucosal surfaces, skin and cornea. Mice are widely used as an animal model to investigate HSV pathogenesis, since the murine homologs of nectin-1 and HVEM also act as efficient receptors and the presence of either nectin-1 or HVEM is sufficient for disease development in mice [2]. Although gD interactions with HVEM or nectin-1 have been studied extensively in different cell lines, the contributions of the individual receptors to uptake into tissue are not well understood.We established an ex vivo infection model of murine epidermal sheets to analyze the contribution of the receptors, and used an experimental setting that enables the virus to enter via the basal layer of the epidermis. Infection studies in HVEM-or nectin-1-deficient epidermis identified nectin-1 as the major receptor in the epidermal sheets, while HVEM had a more limited role [3]. Keratinocytes are the major cell type in the epidermis and when cultured murine primary keratinocytes that expressed neither HVEM nor nectin-1 were examined, almost no infected cells were observed [3]. Since the epidermis represents only the outermost layer of skin, we also addressed the contribution of nectin-1 and HVEM as receptors in the underlying dermis. Fibroblasts are the major resident cell type of the dermis. When we infected murine primary dermal fibroblasts which were deficient in nectin-1, infection was slower, suggesting that HVEM is a less efficient receptor. In the absence of both HVEM and nectin-1, infection was severely delayed resulting in greatly reduced viral spreading and virus production [4]. In contrast to cultured keratinocytes, there was residual infection suggesting the presence of a further, rather inefficient receptor.Comparison of the two major cell types of skin, keratinocytes in the epidermis and fibroblasts in the underlying dermis, demonstrated that nectin-1 is less highly expressed on fibroblasts than on keratinocytes. In contrast, HVEM is present on nearly all fibroblasts but only expressed on a few keratinocytes in epidermis. Interestingly, these expression levels do not appear to correlate with their effectiveness a...

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Initial Contact: The First Steps in Herpesvirus Entry

Azab

Osterrieder

2017

Advances in Anatomy, Embryology and Cell Biology

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The entry process of herpesviruses into host cells is complex and highly variable. It involves a sequence of well-orchestrated events that begin with virus attachment to glycan-containing proteinaceous structures on the cell surface. This initial contact tethers virus particles to the cell surface and results in a cascade of molecular interactions, including the tight interaction of viral envelope glycoproteins to specific cell receptors. These interactions trigger intracellular signaling and finally virus penetration after fusion of the viral envelope with cellular membranes. Based on the engaged cellular receptors and co-receptors, and the subsequent signaling cascades, the entry pathway will be decided on the spot. A number of viral glycoproteins and many cellular receptors and molecules have been identified as players in one or several of these events during virus entry. This chapter will review viral glycoproteins, cellular receptors and signaling cascades associated with the very first interactions of herpesviruses with their target cells.

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

Cited by 34 publications

References 44 publications

Herpes Simplex Virus 1 Enters Human Keratinocytes by a Nectin-1-Dependent, Rapid Plasma Membrane Fusion Pathway That Functions at Low Temperature

Herpes Simplex Virus 1 Enters Human Keratinocytes by a Nectin-1-Dependent, Rapid Plasma Membrane Fusion Pathway That Functions at Low Temperature

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Initial Contact: The First Steps in Herpesvirus Entry

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