• Korean Journal of Microbiology
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• v.36 no.4
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• pp.306-311
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• 2000

Infection of Vero cells with herpes simplex virus type-1 (HSV-1) resulted in a series of changes in intra-cellular free calcium concentration $([Ca^{2+}]_i)$. A significant and maximal decrease $[Ca^{2+}]_i$ was observed at 4 hours postinfection (hr p.i.) in HSV-1-infected in Vero cells. Inactivation of HSV-1 with UV irradiation and heat treatment abolished HSV-1-induced decrease in $[Ca^{2+}]_i$ at 4 hr p.i. in Vero cells. And the degree of the decrease in $[Ca^{2+}]_i$ was dependent on the amount of input virus. Taxol, which stabilizes the polymerization of microtubule blocked HSV-1-induced decrease in $[Ca^{2+}]_i$ at 4 hr p.i., suggesting that microtubule may mediate the transport of HSV-1 nucleocapsid to the nucleus of infected cell. Treatment of HSV-1-infected Vero cells with metabolic inhibitors such as cycloheximide, cordycepin, or acyclovir partially reversed the decrease in $[Ca^{2+}]_i$ at 4 hr p.i.. Thus, it is suggested that HSV-1 induced decrease in $[Ca^{2+}]_i$ at 4 hr p.i. in Vero cells may play an important role in the multiplication of HSV-1.

• Pediatric Infection and Vaccine
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• v.3 no.2
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• pp.207-213
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• 1996

Herpes simplex virus(HSV) infections of the CNS are associated with significant morbidity and mortality even when appropriate antiviral therapy is administered. HSV infections of the brain can be subdivided into two categories : neonatal HSV infections, which usually are caused by HSV type 2, and herpes simplex encephalitis(HSE), which occur in patients over 3 months old and is nearly uniformly caused by HSV type 1. The clinical presentation of HSE is one of the focal encephalopathic process associated with altered levels of consciousness, fever, focal seizures and hemiparesis. But because of the lack of pathognomic clinical presentation and diagnostic procedure, the efforts to develop alternative diagnostic procedure have led to the use of new diagnostic technique such as polymerase chain reaction(PCR). We report a case of HSV type 1 encephalitis in 13 month old male infant who presented with altered level of consciousness, fever and focal seizures. With the use of the PCR, HSV-1 DNA was detected in cerebrospinal fluid from the patient. The symptoms and signs of encephalitis subsided by treatment with acyclovir in 14 days.

• The Journal of Korean Society of Virology
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• v.29 no.3
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• pp.175-182
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• 1999

An attractive target for anti-herpes chemotherapy is the herpes simplex virus 1 (HSV-1) protease encoded by the UL26 gene. HSV-1 protease is essential for DNA packaging and virus maturation. To perform high throughput for potent inhibitors, the efficient production of larger amounts of highly purified enzyme and protease activity assay method must be established. In this report, expression in E. coli and purification of the protease gene of HSV-1 strain F was investigated. The protease gene was cloned pET28, and the nucleotide sequence of protease catalytic domain of HSV-1 compared strain F with other strains (KOS and CL101). In these results the F strain was different in base sequence. However, the amino acid sequence was identifical. The HSV-1 protease was purified with His-tagged affinity column. The analysis of HSV-1 protease activity was performed by high performance liquid chromatography.

• YAKHAK HOEJI
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• v.42 no.6
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• pp.607-612
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• 1998

In the course of our search for anti-Herpes simplex virus type I (HSV 1) substances from natural sources, we screened crude drugs for their antiviral activity using SRB assay. T he methanol extract from herb of Brassica rapa (Cruciferae) was found to inhibit HSV-1. Though bioassay-directed fractionation of the extract, anti-HSV-1 agent was isolated by chromatographic separation using Amberlite XAD-4 and Sephadex LH-20. The structure of compound I was elucidated by spectral means including $^1H-^1H$ COSY, HMQC and HMBC to be isorhamnetin 3-O-${\beta}$-D-glucopyranoside (compound I). Compound I was active against HSV-1 with the 50% effective concentration of O.42mg/ml and the 50% cytotoxicity of 5.0mg/ml.

• Korean Journal of Pharmacognosy
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• v.29 no.4
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• pp.277-282
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• 1998

In order to search for anti-Herpes simplex virus type 1(HSV-1) agents, we screened 80 specimens of Korean traditional medicine by SRB assay. The methanol extracts of Thujae orientalis Semen (Cupressaceae) showed strong anti-HSV activity among samples tested. From the butanol fraction of Thujae orientalis Semen anti-HSV-1 agent was isolated by chromatographic separation using Amberlite XAD-4 and Sephadex LH-20. The structure was elucidated by spectroscopic methods, and was identified as ${\beta}-sitosterol$ (compound I). Compound I exhibited anti-HSV-1 activity with $EC_{50}$ of 0.6 mg/ml and $CC_{50}$ of 5.99 mg/ml, respectively.

• Journal of Microbiology and Biotechnology
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• v.10 no.5
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• pp.714-720
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• 2000

Antigens of Herpes simplex virus type 1 (HSV-1) strain F were immunoblotted to identify the most immunodominant one, and the localization of this antigen was then studied using immunoelectron microscopy. The 67.8 kDa antigen appeared to be the most immunodominant one in a mouse model, and it showed randomly scattered and partially clustered distribution on the surface of the virion. The localization study was performed using immunogold with polyclonal anti-HSV-1 sera produced from BALB/c mice, and immunofluorescence demonstrated that the viral products in the HSV-2 infected Vero cells were distributed throughout the infected host cell, however, mainly on the surface of the host membrane.

• Korean Journal of Pharmacognosy
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• v.30 no.1
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• pp.34-39
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• 1999

To search for less toxic antiherpetic agents, the inhibitory effects of twelve kinds of flavonoids including chrysin, quercetin, quercitrin, rutin, fisetin, gossypin, kaempferol, morin, naringenin, naringin, hesperetin and hesperidin on the plaque formation of herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) in Vero cells were examined by plaque reduction assay in vitro. Some flavonoids tested in this study showed potent antiherpetic activity, reducing intracellular replication of herpes simplex viruses when Vero cell monolayers were infected and subsequently cultured in medium containing flavonoids. Naringenin showed the most potent antiviral activity against HSV-1 with selectivity index (SI) of 19.1 and hesperetin showed the most potent antiviral activity against HSV-2 with SI of 9.8. These results suggest that some flavonoids may be a potential therapeutic agent for the treatment and prevention of herpes simplex virus infections.

• The Korean Journal of Pharmacology
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• v.22 no.2
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• pp.123-127
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• 1986

n-Butyrate (n-BT A) increased the rate and number of infectious units produced in the in vitro reactivation of latent herpes simplex virus. While the mechanism of action of n-BT A is obscure, a continuous presence of n-BT A is necessary for its inductive effect.

• The Journal of the Korean Society for Microbiology
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• v.35 no.1
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• pp.1-8
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• 2000

In order to explore the potential of ascorbic acid supplementation for the prevention and treatment of herpes simplex viral diseases, plaque reduction assays were performed. Ascorbic acid as well as copper chloride/ferric chloride were added to wells containing Vero cells infected with herpes simplex virus type 1 (HSV-1), and the infectivity of HSV-1 was determined. Since copper and iron are major transition metals in human plasma, near the normal human plasma concentrations of them were used for experiments. When Cu(II) and Fe(III) were applied, there were no significant differences between virus control and Cu(II)/Fe(III)-treated groups. But, when appropriate concentrations of ascorbic acid were added to wells, meaningful differences between control and ascorbate-treated groups were found. In the presence of Cu(II)/Fe(III) at $5.8/3.7\;{\mu}M$, 72-h treatment with ascorbate at $50\;{\mu}M$ reduced HSV-1 infections to $10.77%{\pm}4.25%$ (P < 0.001) and $500\;{\mu}M$ did to $3.06%{\pm}1.62%$ (P < 0.001). Moreover, the cytotoxicities for Vero cells at those concentrations were insignificant (P > 0.05). Current recommended dietary allowance (RDA) of ascorbic acid is 60 mg/day, and the oral intake of 60 mg/day of ascorbic acid yields plasma ascorbic acid at 45 to $58\;{\mu}M$ in a healthy adult man. Therefore, the results of this study suggest that the maintenance of appropriate level (more than $50\;{\mu}M$) of ascorbic acid in human plasma by appropriate amount (more than the RDA) of ascorbic acid supplementation may be helpful for the prevention and treatment of diseases caused by HSV -1 in an adult man. In addition, this study also suggests that ascorbic acid may be useful for the prophylaxis of fatal HSV-1 infections in neonates and the prevention of HSV-1 reactivation in immunocompromised hosts.

• Journal of Microbiology and Biotechnology
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• v.11 no.2
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• pp.173-178
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• 2001

A monoclonal antibody (mAb) to the glucoprotein D (gD) of Herpes simplex virus type 2 (HSV-2) was successfully generated by hybridoma technology and characterized. The mAb, SKS2v, recognized a gD antigen with an apparent molecular mass of 60kDa in a Western blot analysis. The isotype was determined by a sandwich ELISA to be IgG2a. HSV-2 exhibited major antigens of 36, 43, 46, 47, 60, 69, 81, 96, 109, 112, 159, and 227 kDa among 25 protein profiles in SDS-PAGE, and among these antigens, those of 60, 112, 125, and 227 kDa were immunodominant in a Western blot analysis using antisera, thereby indicating that they play a role in inducing neutralizing antibodies in HSV-2 infection. When reacted with Vero cells infected with HSV-1 and HSV-2 SKSv2 showed a reactivity to the surface of the infected cells, and a gD antigen of 60 kDa appeared to be expressed in both types of HSV.