scholarly journals Specific Association of Glycoprotein B with Lipid Rafts during Herpes Simplex Virus Entry

2003 ◽  
Vol 77 (17) ◽  
pp. 9542-9552 ◽  
Author(s):  
Florent C. Bender ◽  
J. Charles Whitbeck ◽  
Manuel Ponce de Leon ◽  
Huan Lou ◽  
Roselyn J. Eisenberg ◽  
...  
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Lipid Rafts ◽  
Ebola Virus ◽  
Virus Entry ◽  
Vero Cells ◽  
Inhibitory Effect ◽  
Membrane Lipid ◽  
Dependent Manner ◽  
Simplex Virus

ABSTRACT Herpes simplex virus (HSV) entry requires the interaction of glycoprotein D (gD) with a cellular receptor such as herpesvirus entry mediator (HVEM or HveA) or nectin-1 (HveC). However, the fusion mechanism is still not understood. Since cholesterol-enriched cell membrane lipid rafts are involved in the entry of other enveloped viruses such as human immunodeficiency virus and Ebola virus, we tested whether HSV entry proceeds similarly. Vero cells and cells expressing either HVEM or nectin-1 were treated with cholesterol-sequestering drugs such as methyl-β-cyclodextrin or nystatin and then exposed to virus. In all cases, virus entry was inhibited in a dose-dependent manner, and the inhibitory effect was fully reversible by replenishment of cholesterol. To examine the association of HVEM and nectin-1 with lipid rafts, we analyzed whether they partitioned into nonionic detergent-insoluble glycolipid-enriched membranes (DIG). There was no constitutive association of either receptor with DIG. Binding of soluble gD or virus to cells did not result in association of nectin-1 with the raft-containing fractions. However, during infection, a fraction of gB but not gC, gD, or gH associated with DIG. Similarly, when cells were incubated with truncated soluble glycoproteins, soluble gB but not gC was found associated with DIG. Together, these data favor a model in which HSV uses gB to rapidly mobilize lipid rafts that may serve as a platform for entry and cell signaling. It also suggests that gB may interact with a cellular molecule associated with lipid rafts.

scholarly journals Roles for Endocytosis and Low pH in Herpes Simplex Virus Entry into HeLa and Chinese Hamster Ovary Cells

2003 ◽  
Vol 77 (9) ◽  
pp. 5324-5332 ◽  
Author(s):  
Anthony V. Nicola ◽  
Anna M. McEvoy ◽  
Stephen E. Straus
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Cell Surface ◽  
Vero Cell ◽  
Cell Lines ◽  
Vero Cells ◽  
Low Ph ◽  
Dependent Manner ◽  
Cho Cell ◽  
Simplex Virus

ABSTRACT Herpes simplex virus (HSV) infection of many cultured cells, e.g., Vero cells, can be initiated by receptor binding and pH-neutral fusion with the cell surface. Here we report that a major pathway for HSV entry into the HeLa and CHO-K1 cell lines is dependent on endocytosis and exposure to a low pH. Enveloped virions were readily detected in HeLa or receptor-expressing CHO cell vesicles by electron microscopy at <30 min postinfection. As expected, images of virus fusion with the Vero cell surface were prevalent. Treatment with energy depletion or hypertonic medium, which inhibits endocytosis, prevented uptake of HSV from the HeLa and CHO cell surface relative to uptake from the Vero cell surface. Incubation of HeLa and CHO cells with the weak base ammonium chloride or the ionophore monensin, which elevate the low pH of organelles, blocked HSV entry in a dose-dependent manner. Noncytotoxic concentrations of these agents acted at an early step during infection by HSV type 1 and 2 strains. Entry mediated by the HSV receptor HveA, nectin-1, or nectin-2 was also blocked. As analyzed by fluorescence microscopy, lysosomotropic agents such as the vacuolar H+-ATPase inhibitor bafilomycin A1 blocked the delivery of virus capsids to the nuclei of the HeLa and CHO cell lines but had no effect on capsid transport in Vero cells. The results suggest that HSV can utilize two distinct entry pathways, depending on the type of cell encountered.

scholarly journals Functional Region IV of Glycoprotein D from Herpes Simplex Virus Modulates Glycoprotein Binding to the Herpesvirus Entry Mediator

1998 ◽  
Vol 72 (9) ◽  
pp. 7091-7098 ◽  
Author(s):  
Ann H. Rux ◽  
Sharon H. Willis ◽  
Anthony V. Nicola ◽  
Wangfang Hou ◽  
Charline Peng ◽  
...  
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Virus Entry ◽  
Vero Cells ◽  
Optical Biosensor ◽  
Glycoprotein D ◽  
Functional Region ◽  
Functional Regions ◽  
Simplex Virus ◽  
Herpesvirus Entry Mediator

Glycoprotein D (gD) of herpes simplex virus (HSV) is essential for virus entry and has four functional regions (I to IV) important for this process. We previously showed that a truncated form of a functional region IV variant, gD1(Δ290-299t), had an enhanced ability to block virus entry and to bind to the herpesvirus entry mediator (HveAt; formerly HVEMt), a cellular receptor for HSV. To explore this phenotype further, we examined other forms of gD, especially ones with mutations in region IV. Variant proteins with deletions of amino acids between 277 and 300 (region IV), as well as truncated forms lacking C-terminal residues up to amino acid 275 of gD, were able to block HSV entry into Vero cells 1 to 2 logs better than wild-type gD1(306t). In contrast, gD truncated at residue 234 did not block virus entry into Vero cells. Using optical biosensor technology, we recently showed that gD1(Δ290-299t) had a 100-fold-higher affinity for HveAt than gD1(306t) (3.3 × 10−8 M versus 3.2 × 10−6 M). Here we found that the affinities of other region IV variants for HveAt were similar to that of gD1(Δ290-299t). Thus, the affinity data follow the same hierarchy as the blocking data. In each case, the higher affinity was due primarily to a faster k onrather than to a slower k off. Therefore, once the gDt-HveAt complex formed, its stability was unaffected by mutations in or near region IV. gD truncated at residue 234 bound to HveAt with a lower affinity (2.0 × 10−5 M) than did gD1(306t) due to a more rapid k off. These data suggest that residues between 234 and 275 are important for maintaining stability of the gDt-HveAt complex and that functional region IV is important for modulating the binding of gD to HveA. The binding properties of any gD1(234t)-receptor complex could account for the inability of this form of gDt to block HSV infection.

scholarly journals Comparative Study of Mechanisms of Herpes Simplex Virus Inactivation by Sodium Lauryl Sulfate and n-Lauroylsarcosine

2002 ◽  
Vol 46 (9) ◽  
pp. 2933-2942 ◽  
Author(s):  
Jocelyne Piret ◽  
Sylvie Roy ◽  
Mylène Gagnon ◽  
Sébastien Landry ◽  
André Désormeaux ◽  
...  
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Sodium Lauryl Sulfate ◽  
Cultured Cells ◽  
Vero Cells ◽  
Dependent Manner ◽  
Lauryl Sulfate ◽  
Concentration Dependent Manner ◽  
Simplex Virus

ABSTRACT The mechanisms of herpes simplex virus (HSV) inactivation by sodium lauryl sulfate (SLS) and n-lauroylsarcosine (LS), two anionic surfactants with protein denaturant potency, have been evaluated in cultured cells. Results showed that pretreatment of HSV type 1 (HSV-1) strain F and HSV-2 strain 333 with either surfactant inhibited, in a concentration- and time-dependent manner, their infectivities on Vero cells. SLS was a more potent inhibitor of HSV-2 strain 333 infectivity than LS with respect to the concentration (4.8-fold lower) and time (2.4-fold shorter) required to completely inactivate the virus. No inhibition of both herpesvirus strains infectivities was observed when Vero cells were pretreated with either surfactant. LS prevented the binding of HSV-2 strain 333 to cells without affecting the stable attachment and the rate of penetration into cells, whereas SLS exerted the opposite effect. Both SLS and LS inhibited, in a concentration-dependent manner, the HSV-2 strain 333-induced cytopathic effect, probably by affecting newly synthesized virions that come into contact with surfactant molecules present in culture medium. The pretreatment of HSV-2 strain 333 with specific combinations of SLS and LS concentrations inhibited the viral infectivity in a synergistic manner and resulted in only a small increase in their toxicities for exponentially growing Vero cells compared with that caused by each compound alone. Taken together, these results suggest that SLS and LS, alone or combined, could represent potent candidates as microbicides in topical vaginal formulations to prevent the transmission of herpes and possibly other pathogens that cause sexually transmitted diseases, including human immunodeficiency virus type 1.

scholarly journals Herpes Simplex Virus Glycoproteins H/L Bind to Cells Independently of αVβ3 Integrin and Inhibit Virus Entry, and Their Constitutive Expression Restricts Infection

2010 ◽  
Vol 84 (8) ◽  
pp. 4013-4025 ◽  
Author(s):  
Tatiana Gianni ◽  
Arianna Cerretani ◽  
Rebecca DuBois ◽  
Stefano Salvioli ◽  
Scott S. Blystone ◽  
...  
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Virus Entry ◽  
Constitutive Expression ◽  
K562 Cells ◽  
Protein S ◽  
Αvβ3 Integrin ◽  
Dependent Manner ◽  
Infection Inhibition ◽  
Simplex Virus

ABSTRACT Herpes simplex virus (HSV) fusion with cells requires the gD, gB, and gH/gL glycoprotein quartet. gD serves as a receptor binding glycoprotein. gB and gH/gL execute fusion in an as-yet-unclear manner. To better understand the role of gH/gL in HSV entry, we produced a soluble version of gH/gL carrying a One-STrEP tag (gHt.st/gL). Previous findings implicated integrins as possible ligands to gH/gL (C. Parry et al., J. Gen. Virol. 86:7-10, 2005). We report that (i) gHt.st/gL bound a number of cells in a dose-dependent manner at concentrations similar to those required for the binding of soluble gB or gD. (ii) gHt.st/gL inhibited HSV entry at the same concentrations required for binding. It also inhibited cell-cell fusion in transfected cells. (iii) The absence of β3 integrin did not prevent the binding of gHt.st/gL to CHO cells and infection inhibition. Conversely, integrin-negative K562 cells did not acquire the ability to bind gHt.st/gL when hyperexpressing αVβ3 integrin. (iv) Constitutive expression of wild-type gH/gL (wt-gH/gL) restricted infection in all of the cell lines tested, a behavior typical of glycoproteins which bind cellular receptors. The extent of restriction broadly paralleled the efficiency of gH/gL transfection. RGD motif mutant gH/gL could not be differentiated from wt-gH with respect to restriction of infection. Cumulatively, the present results provide several lines of evidence that HSV gH/gL interacts with a cell surface cognate protein(s), that this protein is not necessarily an αVβ3 integrin, and that this interaction is required for the process of virus entry/fusion.

Inhibitory Effect of Polyionic Compounds on the Adsorption of Herpes Simplex Virus Type 1 (KOS)

1997 ◽  
Vol 8 (1) ◽  
pp. 32-37 ◽  
Author(s):  
Y-W Yang ◽  
J-C Yang
Keyword(s):  
Molecular Weight ◽  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Vero Cells ◽  
Cellular Membrane ◽  
Inhibitory Effect ◽  
The Difference ◽  
Simplex Virus ◽  
Hsv 1

The polyionic compounds, including dextran sulphates and poly-L-lysines, were evaluated for their inhibitory effect on the cytopathic effect of herpes simplex virus (HSV-1, KOS) in vitro. The anti-HSV activities of poly-L-lysines were found to increase with increasing molecular weight of the polymers. Both dextran sulphates and poly-L-lysines were found to block adsorption of HSV-1 to Vero cells. The inhibitory effect of adsorption of [3H] labelled virus was related to the molecular weight of the polymers. Polymers of higher molecular weight were found to be more effective than the lower molecular weight samples in inhibiting virus adsorption. The results from the microelectrophoresis measurements demonstrated that poly-L-lysines adsorb and confer positive charges on the Vero cells. Dextran sulphates, on the other hand, may adsorb onto the HSV-1 membrane surfaces instead of binding onto the cell membranes and interfere with adsorption of virions to the cells. The inhibitory effects of these polymers on viral cytopathogenic effect were probably attributable to the electrostatic and steric hindrance effects exerted by the polymers as reflected in the difference in zeta potential of cellular membrane treated with these compounds.

scholarly journals ent-Epiafzelechin-(4α→8)-epiafzelechin extracted from Cassia javanica inhibits herpes simplex virus type 2 replication

2006 ◽  
Vol 55 (2) ◽  
pp. 201-206 ◽  
Author(s):  
Hua-Yew Cheng ◽  
Chien-Min Yang ◽  
Ta-Chen Lin ◽  
Den-En Shieh ◽  
Chun-Ching Lin
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Inhibitory Effect ◽  
Human Populations ◽  
Dependent Manner ◽  
Virus Attachment ◽  
Life Threatening ◽  
Simplex Virus

Herpes simplex virus (HSV) is a ubiquitous organism that causes infections in human populations throughout the world. It causes a variety of diseases ranging in severity from mild to life-threatening. In this study, ent-epiafzelechin-(4α→8)-epiafzelechin (EEE) extracted from the fresh leaves of Cassia javanica L. agnes de Wit (Leguminosae) was investigated for its in vitro anti-HSV-2 activity using XTT and plaque reduction assays. Results showed that EEE inhibited HSV-2 replication in a dose-dependent manner. The IC50 value was 83·8±10·9 and 166·8±12·9 μM for XTT and plaque reduction assays, respectively. EEE did not affect the viability and the proliferation of cells at antiviral concentrations. Mechanistic studies demonstrated that EEE prevented HSV-2 from penetrating the cell and also interfered with HSV-2 replication at the late stage of its life cycle. It also disturbed virus attachment but the inhibitory effect was minor. In summary, the conclusion of this study was that EEE exhibits various modes of action in suppressing HSV-2 multiplication.

scholarly journals Stable Association of Herpes Simplex Virus with Target Membranes Is Triggered by Low pH in the Presence of the gD Receptor, HVEM

2006 ◽  
Vol 80 (8) ◽  
pp. 3773-3780 ◽  
Author(s):  
J. Charles Whitbeck ◽  
Yi Zuo ◽  
Richard S. B. Milne ◽  
Gary H. Cohen ◽  
Roselyn J. Eisenberg
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Binding Assay ◽  
Virus Entry ◽  
Dependent Manner ◽  
Acidic Ph ◽  
Lower Efficiency ◽  
Ph Dependent ◽  
Stable Association ◽  
Simplex Virus

ABSTRACT Using a liposome-binding assay, we investigated the requirements for activation of herpes simplex virus (HSV) into a state capable of membrane interaction. Virions were mixed with liposomes along with the ectodomain of one of three gD receptors (HVEMt, nectin-1t, or nectin-2t) and incubated under different pH and temperature conditions. Virions failed to associate with liposomes in the presence of nectin-1 or nectin-2 at any temperature or pH tested. In contrast, HVEMt triggered association of HSV with liposomes at pH 5.3 or 5.0 when incubated at 37°C, suggesting that HVEM binding and mildly acidic pH at a physiological temperature provide coactivation signals, allowing virus association with membranes. Virions incubated with HVEMt at 37°C without liposomes rapidly lost infectivity upon exposure to pH 5.0, suggesting that these conditions lead to irreversible virus inactivation in the absence of target membranes. Consistent with the idea that soluble receptor molecules provide a trigger for HSV entry, HVEMt promoted virus entry into receptor-deficient CHO K1 cells. However, in B78H1 cells, HVEMt promoted virus entry with markedly lower efficiency. Interestingly, HSV entry into receptor-bearing CHO K1 cells has been shown to proceed via a pH-dependent manner, whereas HSV entry into receptor-bearing B78H1 cells is pH independent. Based on these observations, we propose that the changes triggered by HVEM and mildly acidic pH that allow liposome association are similar or identical to changes that occur during pH-dependent HSV entry.

scholarly journals 17-β Estradiol Promotion of Herpes Simplex Virus Type 1 Reactivation Is Estrogen Receptor Dependent

2009 ◽  
Vol 84 (1) ◽  
pp. 565-572 ◽  
Author(s):  
Rodolfo D. Vicetti Miguel ◽  
Brian S. Sheridan ◽  
Stephen A. K. Harvey ◽  
Robert S. Schreiner ◽  
Robert L. Hendricks ◽  
...  
Keyword(s):  
Estrogen Receptor ◽  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Viral Reactivation ◽  
Hormonal Contraceptives ◽  
Viral Gene ◽  
Dependent Manner ◽  
Ovariectomized Mice ◽  
Latently Infected ◽  
Simplex Virus

ABSTRACT Correlations between estrogen and herpes simplex virus (HSV) reactivation from latency have been suggested by numerous clinical reports, but causal associations are not well delineated. In a murine HSV-1 corneal infection model, we establish 17-β estradiol (17-βE) treatment of latently infected ovariectomized mice induces viral reactivation, as demonstrated by increased viral load and increased immediate-early viral gene expression in the latently infected trigeminal ganglia (TG). Interestingly, the increased HSV reactivation occurred in the absence of inhibition of viral specific CD8+ T-cell effector function. 17-βE administration increased HSV reactivation in CD45+ cell-depleted TG explant cultures, providing further support that leukocyte-independent effects on latently infected neurons were responsible for the increased reactivation. The drug-induced increases in HSV copy number were not recapitulated upon in vivo treatment of latently infected estrogen receptor alpha-deficient mice, evidence that HSV reactivation promoted by 17-βE was estrogen receptor dependent. These findings provide additional framework for the emerging conceptualization of HSV latency as a dynamic process maintained by complex interactions among multiple cooperative and competing host, viral, and environmental forces. Additional research is needed to confirm whether pregnancy or hormonal contraceptives containing 17-βE also promote HSV reactivation from latency in an estrogen receptor-dependent manner.

Sign in / Sign up

Export Citation Format

Share Document