A herpes simplex virus 1 recombinant lacking the glycoprotein G coding sequences is defective in entry through apical surfaces of polarized epithelial cells in culture and in vivo

Herpes simplex virus 1 (HSV-1) is a human pathogen that leads to recurrent facial-oral lesions. Its 152-kb genome is organized in two covalently linked segments, each composed of a unique sequence flanked by inverted repeats. Replication of the HSV-1 genome produces concatemeric molecules in which homologous recombination events occur between the inverted repeats. This mechanism leads to four genome isomers (termed P, IS, IL, and ILS) that differ in the relative orientations of their unique fragments. Molecular combing analysis was performed on DNA extracted from viral particles and BSR, Vero, COS-7, and Neuro-2a cells infected with either strain SC16 or KOS of HSV-1, as well as from tissues of experimentally infected mice. Using fluorescence hybridization, isomers were repeatedly detected and distinguished and were accompanied by a large proportion of noncanonical forms (40%). In both cell and viral-particle extracts, the distributions of the four isomers were statistically equivalent, except for strain KOS grown in Vero and Neuro-2a cells, in which P and IS isomers were significantly overrepresented. In infected cell extracts, concatemeric molecules as long as 10 genome equivalents were detected, among which, strikingly, the isomer distributions were equivalent, suggesting that any such imbalance may occur during encapsidation.In vivo, for strain KOS-infected trigeminal ganglia, an unbalanced distribution distinct from the onein vitrowas observed, along with a considerable proportion of noncanonical assortment.

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Herpes Simplex Virus Entry by a Nonconventional Endocytic Pathway

Journal of Virology ◽

10.1128/jvi.01910-20 ◽

2020 ◽

Vol 94 (24) ◽

Author(s):

Giulia Tebaldi ◽

Suzanne M. Pritchard ◽

Anthony V. Nicola

Keyword(s):

Herpes Simplex Virus ◽

Herpes Simplex ◽

Epithelial Cells ◽

Retrograde Transport ◽

Endocytic Pathway ◽

Host Cells ◽

Herpes Simplex Virus 1 ◽

Simplex Virus ◽

Golgi Network ◽

Hsv 1

ABSTRACT Herpes simplex virus 1 (HSV-1) causes significant morbidity and mortality in humans worldwide. HSV-1 enters epithelial cells via an endocytosis mechanism that is low-pH dependent. However, the precise intracellular pathway has not been identified, including the compartment where fusion occurs. In this study, we utilized a combination of molecular and pharmacological approaches to better characterize HSV entry by endocytosis. HSV-1 entry was unaltered in both cells treated with small interfering RNA (siRNA) to Rab5 or Rab7 and cells expressing dominant negative forms of these GTPases, suggesting entry is independent of the conventional endo-lysosomal network. The fungal metabolite brefeldin A (BFA) and the quinoline compound Golgicide A (GCA) inhibited HSV-1 entry via beta-galactosidase reporter assay and impaired incoming virus transport to the nuclear periphery, suggesting a role for trans-Golgi network (TGN) functions and retrograde transport in HSV entry. Silencing of Rab9 or Rab11 GTPases, which are involved in the retrograde transport pathway, resulted in only a slight reduction in HSV infection. Together, these results suggest that HSV enters host cells by an intracellular route independent of the lysosome-terminal endocytic pathway. IMPORTANCE Herpes simplex virus 1 (HSV-1), the prototype alphaherpesvirus, is ubiquitous in the human population and causes lifelong infection that can be fatal in neonatal and immunocompromised individuals. HSV enters many cell types by endocytosis, including epithelial cells, the site of primary infection in the host. The intracellular itinerary for HSV entry remains unclear. We probed the potential involvement of several Rab GTPases in HSV-1 entry and suggest that endocytic entry of HSV-1 is independent of the canonical lysosome-terminal pathway. A nontraditional endocytic route may be employed, such as one that intersects with the trans-Golgi network (TGN). These results may lead to novel targets for intervention.

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Restoring Herpesvirus Entry Mediator (HVEM) Immune Function in HVEM−/− Mice Rescues Herpes Simplex Virus 1 Latency and Reactivation Independently of Binding to Glycoprotein D

Journal of Virology ◽

10.1128/jvi.00700-20 ◽

2020 ◽

Vol 94 (16) ◽

Author(s):

Kati Tormanen ◽

Shaohui Wang ◽

Ujjaldeep Jaggi ◽

Homayon Ghiasi

Keyword(s):

Herpes Simplex Virus ◽

Herpes Simplex ◽

Immune Function ◽

Interleukin 2 ◽

Glycoprotein D ◽

Herpes Simplex Virus 1 ◽

Simplex Virus ◽

Hsv 1

ABSTRACT The immune modulatory protein herpes virus entry mediator (HVEM) is one of several cellular receptors used by herpes simplex virus 1 (HSV-1) for cell entry. HVEM binds to HSV-1 glycoprotein D (gD) but is not necessary for HSV-1 replication in vitro or in vivo. Previously, we showed that although HSV-1 replication was similar in wild-type (WT) control and HVEM−/− mice, HSV-1 does not establish latency or reactivate effectively in mice lacking HVEM, suggesting that HVEM is important for these functions. It is not known whether HVEM immunomodulatory functions contribute to latency and reactivation or whether its binding to gD is necessary. We used HVEM−/− mice to establish three transgenic mouse lines that express either human WT HVEM or human or mouse HVEM with a point mutation that ablates its ability to bind to gD. Here, we show that HVEM immune function, not its ability to bind gD, is required for WT levels of latency and reactivation. We further show that HVEM binding to gD does not affect expression of the HVEM ligands BTLA, CD160, or LIGHT. Interestingly, our results suggest that binding of HVEM to gD may contribute to efficient upregulation of CD8α but not PD1, TIM-3, CTLA4, or interleukin 2 (IL-2). Together, our results establish that HVEM immune function, not binding to gD, mediates establishment of latency and reactivation. IMPORTANCE HSV-1 is a common cause of ocular infections worldwide and a significant cause of preventable blindness. Corneal scarring and blindness are consequences of the immune response induced by repeated reactivation events. Therefore, HSV-1 therapeutic approaches should focus on preventing latency and reactivation. Our data suggest that the immune function of HVEM plays an important role in the HSV-1 latency and reactivation cycle that is independent of HVEM binding to gD.

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The Herpes Simplex Virus 1 IgG Fc Receptor Blocks Antibody-Mediated Complement Activation and Antibody-Dependent Cellular Cytotoxicity In Vivo

Journal of Virology ◽

10.1128/jvi.02509-10 ◽

2011 ◽

Vol 85 (7) ◽

pp. 3239-3249 ◽

Cited By ~ 44

Author(s):

J. M. Lubinski ◽

H. M. Lazear ◽

S. Awasthi ◽

F. Wang ◽

H. M. Friedman

Keyword(s):

Herpes Simplex Virus ◽

Herpes Simplex ◽

Complement Activation ◽

Fc Receptor ◽

Cellular Cytotoxicity ◽

Antibody Dependent Cellular Cytotoxicity ◽

Herpes Simplex Virus 1 ◽

Simplex Virus

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Effects of Phosphorylation of Herpes Simplex Virus 1 Envelope Glycoprotein B by Us3 Kinase In Vivo and In Vitro

Journal of Virology ◽

10.1128/jvi.01447-09 ◽

2009 ◽

Vol 84 (1) ◽

pp. 153-162 ◽

Cited By ~ 30

Author(s):

Takahiko Imai ◽

Ken Sagou ◽

Jun Arii ◽

Yasushi Kawaguchi

Keyword(s):

Herpes Simplex Virus ◽

Herpes Simplex ◽

Viral Replication ◽

Envelope Glycoprotein ◽

Critical Role ◽

Glycoprotein B ◽

Herpes Simplex Virus 1 ◽

Simplex Virus ◽

Hsv 1

ABSTRACT We recently reported that the herpes simplex virus 1 (HSV-1) Us3 protein kinase phosphorylates threonine at position 887 (Thr-887) in the cytoplasmic tail of envelope glycoprotein B (gB) (A. Kato, J. Arii, I. Shiratori, H. Akashi, H. Arase, and Y. Kawaguchi, J. Virol. 83:250-261, 2009; T. Wisner, C. C. Wright, A. Kato, Y. Kawaguchi, F. Mou, J. D. Baines, R. J. Roller and D. C. Johnson, J. Virol. 83:3115-3126, 2009). In the studies reported here, we examined the effect(s) of this phosphorylation on viral replication and pathogenesis in vivo and present data showing that replacement of gB Thr-887 by alanine significantly reduced viral replication in the mouse cornea and development of herpes stroma keratitis and periocular skin disease in mice. The same effects have been reported for mice infected with a recombinant HSV-1 carrying a kinase-inactive mutant of Us3. These observations suggested that Us3 phosphorylation of gB Thr-887 played a critical role in viral replication in vivo and in HSV-1 pathogenesis. In addition, we generated a monoclonal antibody that specifically reacted with phosphorylated gB Thr-887 and used this antibody to show that Us3 phosphorylation of gB Thr-887 regulated subcellular localization of gB, particularly on the cell surface of infected cells.

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Herpes simplex virus type 1 exhibits a tropism for basal entry in polarized epithelial cells

Journal of General Virology ◽

10.1099/vir.0.19226-0 ◽

2003 ◽

Vol 84 (9) ◽

pp. 2473-2484 ◽

Cited By ~ 37

Author(s):

Mario Schelhaas ◽

Matthias Jansen ◽

Ingo Haase ◽

Dagmar Knebel-Mörsdorf

Keyword(s):

Herpes Simplex Virus ◽

Herpes Simplex ◽

Epithelial Cells ◽

Virus Type ◽

Herpes Simplex Virus Type ◽

Polarized Epithelial Cells ◽

Simplex Virus

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RIG-I-Mediated STING Upregulation Restricts Herpes Simplex Virus 1 Infection

Journal of Virology ◽

10.1128/jvi.00748-16 ◽

2016 ◽

Vol 90 (20) ◽

pp. 9406-9419 ◽

Cited By ~ 38

Author(s):

Yiliu Liu ◽

Marie-Line Goulet ◽

Alexandre Sze ◽

Samar Bel Hadj ◽

Sidi Mehdi Belgnaoui ◽

...

Keyword(s):

Herpes Simplex Virus ◽

Immune System ◽

Herpes Simplex ◽

Innate Immune System ◽

Innate Immune ◽

Herpes Simplex Virus 1 ◽

Dna And Rna ◽

Simplex Virus ◽

Hsv 1

ABSTRACTSTING has emerged in recent years as a key player in orchestrating innate immune responses to cytosolic DNA and RNA derived from pathogens. However, the regulation of STING still remains poorly defined. In the present study, we investigated the mechanism of the regulation of STING expression in relation to the RIG-I pathway. Our data show that signaling through RIG-I induces STING expression at both the transcriptional and protein levels in various cell types. STING induction by the RIG-I agonist 5′triphosphorylated RNA (5′pppRNA) was recognized to be a delayed event resulting from an autocrine/paracrine mechanism. Indeed, cotreatment with tumor necrosis factor alpha and type I/II interferon was found to have a synergistic effect on the regulation of STING expression and could be potently decreased by impairing NF-κB and/or STAT1/2 signaling. STING induction significantly contributed to sustainment of the immune signaling cascade following 5′pppRNA treatment. Physiologically, this cross talk between the RNA- and DNA-sensing pathways allowed 5′pppRNA to efficiently block infection by herpes simplex virus 1 (HSV-1) bothin vitroandin vivoin a STING-dependent fashion. These observations demonstrate that STING induction by RIG-I signaling through the NF-κB and STAT1/2 cascades is essential for RIG-I agonist-mediated HSV-1 restriction.IMPORTANCEThe innate immune system represents the first line of defense against invading pathogens. The dysregulation of this system can result in failure to combat pathogens, inflammation, and autoimmune diseases. Thus, precise regulation at each level of the innate immune system is crucial. Recently, a number of studies have established STING to be a central molecule in the innate immune response to cytosolic DNA and RNA derived from pathogens. Here, we describe the regulation of STING via RIG-I-mediated innate immune sensing. We found that STING is synergistically induced via proinflammatory and antiviral cytokine cascades. In addition, we show thatin vivoprotection against herpes simplex virus 1 (HSV-1) by a RIG-I agonist required STING. Our study provides new insights into the cross talk between DNA and RNA pathogen-sensing systems via the control of STING.

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Role for nectin-1 in herpes simplex virus 1 entry and spread in human retinal pigment epithelial cells

FEBS Journal ◽

10.1111/j.1742-4658.2008.06655.x ◽

2008 ◽

Vol 275 (21) ◽

pp. 5272-5285 ◽

Cited By ~ 34

Author(s):

Vaibhav Tiwari ◽

Myung-Jin Oh ◽

Maria Kovacs ◽

Shripaad Y. Shukla ◽

Tibor Valyi-Nagy ◽

...

Keyword(s):

Herpes Simplex Virus ◽

Herpes Simplex ◽

Epithelial Cells ◽

Retinal Pigment Epithelial ◽

Retinal Pigment ◽

Retinal Pigment Epithelial Cells ◽

Herpes Simplex Virus 1 ◽

Pigment Epithelial ◽

Simplex Virus ◽

Pigment Epithelial Cells

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A novel bioluminescent herpes simplex virus 1 for in vivo monitoring of herpes simplex encephalitis

Scientific Reports ◽

10.1038/s41598-021-98047-z ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Olus Uyar ◽

Pier-Luc Plante ◽

Jocelyne Piret ◽

Marie-Christine Venable ◽

Julie Carbonneau ◽

...

Keyword(s):

Herpes Simplex Virus ◽

Herpes Simplex ◽

Cell Culture Supernatant ◽

Herpes Simplex Virus Encephalitis ◽

Herpes Simplex Virus 1 ◽

Simplex Virus ◽

Hsv 1 ◽

Viral Titers

AbstractHerpes simplex virus 1 (HSV-1) is responsible for herpes simplex virus encephalitis (HSE), associated with a 70% mortality rate in the absence of treatment. Despite intravenous treatment with acyclovir, mortality remains significant, highlighting the need for new anti-herpetic agents. Herein, we describe a novel neurovirulent recombinant HSV-1 (rHSV-1), expressing the fluorescent tdTomato and Gaussia luciferase (Gluc) enzyme, generated by the Clustered regularly interspaced short palindromic repeats (CRISPR)—CRISPR-associated protein 9 (Cas9) (CRISPR-Cas9) system. The Gluc activity measured in the cell culture supernatant was correlated (P = 0.0001) with infectious particles, allowing in vitro monitoring of viral replication kinetics. A significant correlation was also found between brain viral titers and Gluc activity in plasma (R2 = 0.8510, P < 0.0001) collected from BALB/c mice infected intranasally with rHSV-1. Furthermore, evaluation of valacyclovir (VACV) treatment of HSE could also be performed by analyzing Gluc activity in mouse plasma samples. Finally, it was also possible to study rHSV-1 dissemination and additionally to estimate brain viral titers by in vivo imaging system (IVIS). The new rHSV-1 with reporter proteins is not only as a powerful tool for in vitro and in vivo antiviral screening, but can also be used for studying different aspects of HSE pathogenesis.

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