Antiviral drug susceptibility of herpes simplex type 1 (HSV-1) strains emerging under the selective pressure of (S)-1-(3-Hydroxy-2-Phosphonylmethoxypropyl)cytosine (HPMPC) in vitro

ABSTRACTDNA polymerases of theHerpesviridaeand bacteriophage RB69 belong to the α-like DNA polymerase family. In spite of similarities in structure and function, the RB69 enzyme is relatively resistant to foscarnet, requiring the mutation V478W in helix N to promote the closed conformation of the enzyme to make it susceptible to the antiviral. Here, we generated recombinant herpes simplex virus 1 (HSV-1) and human cytomegalovirus (HCMV) mutants harboring the revertant in UL30 (W781V) and UL54 (W780V) DNA polymerases, respectively, to further investigate the impact of this tryptophan on antiviral drug susceptibility and viral replicative capacity. The mutation W781V in HSV-1 induced resistance to foscarnet, acyclovir, and ganciclovir (3-, 14-, and 3-fold increases in the 50% effective concentrations [EC50s], respectively). The recombinant HCMV mutant harboring the W780V mutation was slightly resistant to foscarnet (a 1.9-fold increase in the EC50) and susceptible to ganciclovir. Recombinant HSV-1 and HCMV mutants had altered viral replication kinetics. The apparent inhibition constant values of foscarnet against mutant UL30 and UL54 DNA polymerases were 45- and 4.9-fold higher, respectively, than those against their wild-type counterparts. Structural evaluation of the tryptophan position in the UL54 DNA polymerase suggests that the bulkier phenylalanine (fingers domain) and isoleucine (N-terminal domain) could induce a tendency toward the closed conformation greater than that for UL30 and explains the modest effect of the W780V mutation on foscarnet susceptibility. Our results further suggest a role of the tryptophan in helix N in conferring HCMV and especially HSV-1 susceptibility to foscarnet and the possible contribution of other residues localized at the interface between the fingers and N-terminal domains.IMPORTANCEDNA polymerases of theHerpesviridaeand bacteriophage RB69 belong to the α-like DNA polymerase family. However, the RB69 DNA polymerase is relatively resistant to the broad-spectrum antiviral agent foscarnet. The mutation V478W in helix N of the fingers domain caused the enzyme to adopt a closed conformation and to become susceptible to the antiviral. We generated recombinant herpes simplex virus 1 (HSV-1) and human cytomegalovirus (HCMV) mutants harboring the revertant in UL30 (W781V) and UL54 (W780V) DNA polymerases, respectively, to further investigate the impact of this tryptophan on antiviral drug susceptibility. The W781V mutation in HSV-1 induced resistance to foscarnet, whereas the W780V mutation in HCMV slightly decreased drug susceptibility. This study suggests that the different profiles of susceptibility to foscarnet of the HSV-1 and HCMV mutants could be related to subtle conformational changes resulting from the interaction between residues specific to each enzyme that are located at the interface between the fingers and the N-terminal domains.

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Application of Real-Time PCR for Determination of Antiviral Drug Susceptibility of Herpes Simplex Virus

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.46.9.2943-2947.2002 ◽

2002 ◽

Vol 46 (9) ◽

pp. 2943-2947 ◽

Cited By ~ 22

Author(s):

Růžena Stránská ◽

Anton M. van Loon ◽

Merjo Polman ◽

Rob Schuurman

Keyword(s):

Herpes Simplex ◽

Real Time ◽

Real Time Pcr ◽

Antiviral Drug ◽

Drug Susceptibility ◽

Pcr Assay ◽

Simplex Virus ◽

Antiviral Drug Susceptibility ◽

Hsv 1

ABSTRACT A quantitative real-time PCR (TaqMan) assay was developed for determination of antiviral drug susceptibility of herpes simplex virus (HSV). After short-time culture of the virus, the antiviral drug susceptibility of HSV isolates for acyclovir (ACV) was determined by measuring the reduction of the HSV type 1 (HSV-1) DNA levels in culture supernatants using real-time PCR. The 50% inhibitory concentration was reported as the concentration of antiviral drug that reduced the number of HSV-1 DNA copies by 50%. A total of 15 well-characterized ACV-sensitive or -resistant strains and clinical isolates were used for assay evaluation. The new assay with real-time PCR readout permitted rapid (3 days), objective, and reproducible determination of HSV-1 drug susceptibilities with no need for stringent control of initial multiplicity of infection. Furthermore, the real-time PCR assay results showed good correlation (r = 0.86) with those for the plaque reduction assay. In conclusion, the real-time PCR assay described here is a suitable quantitative method for determination of antiviral susceptibility of HSV-1, amenable for use in the routine diagnostic virology laboratory.

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Activity of hydroxyurea (HU) in combination with several antiviral nucleosides against in vitro replication of herpes simplex type 1 (HSV-1) and type 2 (HSV-2) and varicella-zoster virus (VZV)

Antiviral Research ◽

10.1016/0166-3542(95)94866-z ◽

1995 ◽

Vol 26 (3) ◽

pp. A315

Author(s):

R. Snoeck ◽

G. Andrei ◽

E. De Clercq

Keyword(s):

Herpes Simplex ◽

Varicella Zoster Virus ◽

Herpes Simplex Type ◽

Varicella Zoster ◽

In Vitro Replication ◽

Antiviral Nucleosides ◽

Hsv 1

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Entry of Herpes Simplex Virus Type 1 into Primary Sensory Neurons In Vitro Is Mediated by Nectin-1/HveC

Journal of Virology ◽

10.1128/jvi.77.5.3307-3311.2003 ◽

2003 ◽

Vol 77 (5) ◽

pp. 3307-3311 ◽

Cited By ~ 53

Author(s):

Sarah M. Richart ◽

Scott A. Simpson ◽

Claude Krummenacher ◽

J. Charles Whitbeck ◽

Lewis I. Pizer ◽

...

Keyword(s):

Herpes Simplex Virus ◽

Herpes Simplex ◽

Sensory Neurons ◽

Virus Type ◽

Herpes Simplex Virus Type ◽

Primary Cultures ◽

Simplex Virus ◽

Hsv 1

ABSTRACT Primary cultures of rat and mouse sensory neurons were used to study the entry of herpes simplex virus type 1 (HSV-1). Soluble, truncated nectin-1 but not HveA prevented viral entry. Antibodies against nectin-1 also blocked infection of rat neurons. These results indicate that nectin-1 is the primary receptor for HSV-1 infection of sensory neurons.

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Antiviral effect of oryzacystatin, a proteinase inhibitor in rice, against herpes simplex virus type 1 in vitro and in vivo.

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.39.4.846 ◽

1995 ◽

Vol 39 (4) ◽

pp. 846-849 ◽

Cited By ~ 41

Author(s):

H Aoki ◽

T Akaike ◽

K Abe ◽

M Kuroda ◽

S Arai ◽

...

Keyword(s):

Herpes Simplex Virus ◽

Herpes Simplex ◽

Virus Type ◽

Herpes Simplex Virus Type ◽

Rice Seed ◽

Simplex Virus ◽

Hsv 1

Oryzacystatin (OC) is the first-described cystatin originating from rice seed; it consists of two molecular species, OC-I and OC-II, which have antiviral action against poliovirus in vitro (H. Kondo, S. Ijiri, K. Abe, H. Maeda, and S. Arai, FEBS Lett. 299:48-50, 1992). In the experiments reported here, we investigated the effects of OC-I and OC-II on the replication of herpes simplex virus type 1 (HSV-1) in vitro and in vivo. HSV-1 was inoculated onto monolayers of monkey kidney epithelial cells (CV-1 cells) at a multiplicity of infection of 0.1 PFU per cell. After adsorption of the virus onto cells, the cultures were incubated in the presence of either OC-I or OC-II in the concentration range of 1.0 to 300 microM, and the supernatant virus yield was quantitated at 24 h. The effective concentration for 90% inhibition of HSV-1 was 14.8 microM, while a cytotoxic effect on CV-1 cells without infection of HSV-1 was not observed below 500 microM OC-I. Therefore, the apparent in vitro chemotherapeutic index was estimated to be more than 33. In the mouse model of HSV-1-induced keratitis and encephalopathy, topical administration of OC-I to the mouse cornea produced a significant decrease in virus production in the cornea (mean virus yields: 3.11 log10 PFU in the treated group and 4.37 log10 PFU in the control group) and significant improvement in survival rates (P = 0.01). The in vivo antiherpetic effect of OC-I was comparable to that of acyclovir, indicating that topical treatment of HSV-1 infection in humans with OC-I might be possible. Our data also suggest the importance of some thiol proteinases, which may be derived from either the host's cells or HSV-1, during the replication process of HSV-1.

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P176 evaluation of topical brivudin in comparison to aciclovir in experimental cutaneous herpes simplex type 1 (HSV-1) infection in the guinea pig

European Journal of Pharmaceutical Sciences ◽

10.1016/0928-0987(94)90349-2 ◽

1994 ◽

Vol 2 (1-2) ◽

pp. 162

Author(s):

G. Werner ◽

M. Fischer ◽

S. Möller ◽

A. Giachetti

Keyword(s):

Herpes Simplex ◽

Guinea Pig ◽

Herpes Simplex Type ◽

Hsv 1

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UL36p Is Required for Efficient Transport of Membrane-Associated Herpes Simplex Virus Type 1 along Microtubules

Journal of Virology ◽

10.1128/jvi.00225-08 ◽

2008 ◽

Vol 82 (15) ◽

pp. 7388-7394 ◽

Cited By ~ 52

Author(s):

Sara K. Shanda ◽

Duncan W. Wilson

Keyword(s):

Herpes Simplex Virus ◽

Herpes Simplex ◽

Virus Type ◽

Herpes Simplex Virus Type ◽

Anterograde Transport ◽

Tegument Proteins ◽

Simplex Virus ◽

Hsv 1

ABSTRACT Microtubule-mediated anterograde transport is essential for the transport of herpes simplex virus type 1 (HSV-1) along axons, yet little is known regarding the mechanism and the machinery required for this process. Previously, we were able to reconstitute anterograde transport of HSV-1 on microtubules in an in vitro microchamber assay. Here we report that the large tegument protein UL36p is essential for this trafficking. Using a fluorescently labeled UL36 null HSV-1 strain, KΔUL36GFP, we found that it is possible to isolate a membrane-associated population of this virus. Although these viral particles contained normal amounts of tegument proteins VP16, vhs, and VP22, they displayed a 3-log decrease in infectivity and showed a different morphology compared to UL36p-containing virions. Membrane-associated KΔUL36GFP also displayed a slightly decreased binding to microtubules in our microchamber assay and a two-thirds decrease in the frequency of motility. This decrease in binding and motility was restored when UL36p was supplied in trans by a complementing cell line. These findings suggest that UL36p is necessary for HSV-1 anterograde transport.

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In vitro antiviral and immunomodulatory activity of arbidol and structurally related derivatives in herpes simplex virus type 1-infected human keratinocytes (HaCat)

Journal of Medical Microbiology ◽

10.1099/jmm.0.076612-0 ◽

2014 ◽

Vol 63 (11) ◽

pp. 1474-1483 ◽

Cited By ~ 11

Author(s):

Brunella Perfetto ◽

Rosanna Filosa ◽

Vincenza De Gregorio ◽

Antonella Peduto ◽

Annalisa La Gatta ◽

...

Keyword(s):

Herpes Simplex Virus ◽

Herpes Simplex ◽

Virus Type ◽

Herpes Simplex Virus Type ◽

Viral Infections ◽

Antiviral Drug ◽

Immunomodulatory Activity ◽

Simplex Virus

Arbidol (ARB) is an antiviral drug that has broad-spectrum activity against a number of viral infections. To date, there are no specific data regarding its effects against a herpesvirus. Here, the in vitro antiviral effect of ARB and structurally related derivatives were evaluated in HaCat cells on different steps of herpes simplex virus type 1 replication: adsorption, entry and post-entry. The simplified pyrrolidine analogue, 9a2, showed the best antiviral activity in vitro by reducing the plaque numbers by about 50 % instead of 42 % obtained with ARB at the same concentration. Furthermore, we have reported that all tested compounds evaluated for their immunomodulatory activity showed the ability to reduce the viral proteins VP16 and ICP27 and to modify the virus-induced cytokine expression, allowing the host cell a more efficient antiviral response.

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