Antiviral and virucidal activities against herpes simplex viruses of umesu phenolics extracted from Japanese apricot.

Umesu phenolics were obtained from the salt extracts of Japanese apricot (Nanko-mume cultivar of Prunus mume Sieb. et Zucc.) as purified phenolics. The antiviral activities of umesu phenolics obtained were then examined against herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2), enveloped DNA viruses. The phenolics inhibited the multiplication of these viruses when added to the culture media of the infected cells. This inhibition occurred at phenolic concentrations at which they showed no severe cytotoxicity. One-step growth experiments showed that the eclipse period in the HSV-1 multiplication process was extended in the presence of umesu phenolics and that the addition of phenolics after the completion of viral DNA replication did not affect their multiplication. More drastic effects were observed on virucidal activities against HSV-1 and HSV-2; the infectivity decreased to 0.0001 when infected cells were incubated with 3 mg/ml phenolics at 30°C for 5 min. These results demonstrate the antiviral and virucidal activities of umesu phenolics and suggest a potential pharmacological use for these phenolics as a sanitizing or preventive medicine against superficial HSV infections.

[A study of the antiherpetic activity of the chaga mushroom (Inonotus obliquus) extracts in the Vero cells infected with the herpes simplex virus].

The chaga mushroom (Inonotus obliquus) contains a wide range of excellent bioactive compounds. However, limited information exists on the antiviral activity of the compounds extracted from chaga. A number of subfractions of chaga were obtained using different solvents and different procedures. The subfractions of chaga extracted with water, alcohol, alkali were tested for their toxicity for the Vero cell culture and antiviral effect in the Vero cells infected with the Herpes simplex virus (HSV), Type 1. It was shown that most of the subfractions were not toxic for the Vero cells and had protective effect on the Vero cells infected with HSV. The subfraction IV in the concentration 5 microg/ml protected the Vero cells from cytodestructive action of HSV and no viral DNA was detected in infected cells treated with chaga extracts. Best protective effect was observed when compound was added before or within one hour after the Vero cells were infected with HSV.

In vitro anti-herpes simplex viruses and anti-adenoviruses activity of twelve traditionally used medicinal plants in Taiwan.

As an effort to search for new antiviral agents from traditional medicine, the hot water (HW) extract of twelve traditionally used medicinal plants in Taiwan was evaluated for their in vitro anti-herpes simplex viruses (HSV; including HSV-1 and HSV-2) and anti-adenoviruses (ADV; including ADV-3, ADV-8 and ADV-11) activities with a XTT-based colorimetric assay. Results showed that the tested HW extracts exhibited anti-HSV and anti-ADV activities at different magnitudes of potency. Among the twelve medicinal plants, Boussingaultia gracilis var. pseudobaselloides (Basellaceae) and Serissa japonica (Rubiaceae) possessed broad spectrum of antiviral activity. Ardisia squamulosa (Myrsinaceae) and Artemisai princeps var. orientalis (Compositae) were more effective in inhibiting ADV-8 replication than the other four viruses. Cell cytotoxic assay demonstrated that all tested HW extracts had CC50 values higher than their EC50 values. It was concluded that the twelve traditionally used medicinal plants in Taiwan possessed antiviral activity, and some of them merit further investigation.

A randomized, double-blind, placebo-controlled trial of cidofovir gel for the treatment of acyclovir-unresponsive mucocutaneous herpes simplex virus infection in patients with AIDS.

The safety and efficacy of cidofovir gel for treatment of acyclovir-unresponsive herpes simplex virus infections in AIDS patients was evaluated in a randomized, double-blind, multicenter trial. Cidofovir (0.3% or 1%) or placebo gel was applied once daily for 5 days. Ten of 20 cidofovir-treated and none of 10 placebo-treated patients had complete healing or >50% decreased area (P = .008); 30% of cidofovir-treated patients versus 0 placebo recipients had complete healing (P = .031). Viral shedding ceased in 13 (87%) of 15 cidofovir-treated and 0 of 9 placebo-treated patients (P = .00004). For cidofovir-treated patients, median time to complete or good response was 21 days, and median time to negative viral culture was 2 days (P = .025, P = .0001, respectively). Median lesion area decreases were 58% for cidofovir-treated versus 0 for placebo-treated patients (P = .005), and mean pain score changes were -1.84 versus -0.34 (P = .042). Application site reactions occurred in 25% of cidofovir-treated and 20% of placebo-treated patients; none was dose-limiting. Cidofovir therapy provided significant benefits in lesion healing, virologic effect, and pain reduction.

[Efficacy of xiaoxingzhang guttae ophthalmic eye drops in the treatment of experimental herpes simplex keratitis].

The new guttae ophthalmic Xiaoxingzhang (XXZ) was extracted from Radix Actinidiae, a traditional Chinese herbal drug. The 50% inhibition concentration (IC50) of XXZ on type I Herpes Simplex Virus (HSV-1) in virus cell cultures is 165.48-174.73 micrograms/ml. However, XXZ concentrations greater than 400 micrograms/ml did not cause any microscopically visible disruption of vero cells. The efficacy of XXZ in the treatment of experimental Herpes Simplex Keratitis (HSK) in rabbits is higher than that of idoxuridine. The effective doses of XXZ are not toxic to corneal epithelium. The results suggest that XXZ as a new anti-HSV preparation is potentialy useful in the treatment of patients with HSK.

Plaquing efficiency of herpes simplex virus type 1 with a defined agar overlay.

Plaque assay was performed on herpes simplex type 1, using Vero, BHK-21, Hep-2 and LLCMK2 cell lines. The plaquing efficiency was optimal when 60 mm dishes containing Vero cells received an inoculum of 200 microliters with an adsorption time of 90 min. The overlay contained agar and tissue culture medium supplemented with extra vitamins, DEAE-Dextran and magnesium chloride. Virus titer was increased by two fold and plaques were clearly visible within three days of incubation. This modified method was reproducible and is being used routinely in our research and diagnostic laboratories.

Rapid in vivo reactivation of herpes simplex virus in latently infected murine ganglionic neurons after transient hyperthermia.

A rapid and physiologically relevant hyperthermia-based induction procedure has been utilized to develop an in vivo model of induced herpes simplex virus (HSV) reactivation in outbred Swiss Webster mice. This procedure was found to efficiently reactivate latent virus from both trigeminal and lumbosacral ganglia. Examination of the time between hyperthermia and virus production demonstrated that detectable levels of infectious virus were present in ganglia as soon as 14 h posttreatment, with peak percent recoveries at 24 h. These data indicated that the switch from latent to active viral gene transcription occurred rapidly following treatment. Immunohistochemical staining for HSV type 1 antigens revealed rare antigen-positive ganglionic neurons 24 h postinduction. HSV antigens were not detected in any other cell type, and lateral spread of the infection was not observed. This is the first report of the detection of HSV antigens in vivo following induced reactivation in the intact nervous system and demonstrates that the neuron is the site of infectious virus production. In addition, our data strongly suggest that at least some neurons in which HSV antigens are detected during reactivation do not survive. Because the temporal and spatial characteristics of HSV reactivation have been clearly defined, this model is uniquely suited for the molecular dissection of the reactivation process.

Evaluation of 1-(2'-deoxy-2'-fluoro-beta-D-arabinofuranosyl)-5-ethyluracil in a rabbit model of herpetic keratitis.

The nucleoside analog 1-(2'-deoxy-2'-fluoro-beta-D-arabinofuranosyl)-5- ethyluracil (FEAU) was tested in a rabbit model of acute herpetic keratitis and its effectiveness compared with that of acyclovir (ACV). FEAU or ACV was applied topically 3 times daily, beginning 3 days post-HSV-1 inoculation and continued for a period of 7 days. FEAU at a concentration of 1% (w/v) or 3% ACV resulted in significant lessening of the severity of corneal lesions, conjunctivitis, iritis, and corneal clouding at 24 to 48 h after beginning chemotherapy. No toxic reaction was observed in any rabbit eyes treated with either FEAU or ACV. The duration of virus shedding into tear film and colonization of the trigeminal ganglia, however, were not reduced by either FEAU or ACV treatment begun 3 days post-inoculation. Fifty percent effective dose (ED50) of FEAU determinations performed on isolates from tear film and on the virus inoculum in secondary rabbit kidney cultures yielded a range of 4.6-7 microM, with two in vitro resistant isolates having ED50S of greater than or equal to 1500 microM of FEAU. Fifty percent cell growth inhibition for FEAU was 3000 microM at 72 h.

Antiviral effect of mangiferin and isomangiferin on herpes simplex virus.

Using tissue culture technique the present study for the first time indicated the antiviral effect of mangiferin and isomangiferin against type I herpes simplex virus (HSV-I). 4 methods were used for evaluating drug effectiveness (i.e., in vitro drug-on-virus direct action, simultaneous addition of drug-virus-inoculum to cell bottle, virus inoculation preceding drug addition, and drug addition followed by virus inoculation), it was readily found by log determination of HSV-I inhibition that isomangiferin somewhat exceeded such control drugs as acyclovir, idoxuridine, and cyclocytidine in log by 0.27-0.50, and that mangiferin was lower than isomangiferin in log by 0.53. The average plaque reduction rates of mangiferin and isomangiferin were 56.8% and 69.5% respectively. The antiviral effect of mangiferin and isomangiferin was presumably due to their capability of inhibiting virus replication within cells.

Inhibition of herpes simplex virus infection by pine cone antitumor substances.

Several antitumor substances extracted from cones of various pine trees inhibited the plaque formation of Herpes simplex virus types 1 and 2 (HSV-1, HSV-2) strains in African green monkey kidney cells and human adenocarcinoma cells. The 50% effective dose of the most active fraction, Fr. VI (0.3 micrograms/ml) was 0.001 times its 50% cytotoxic dose (greater than 300 micrograms/ml). The anti-HSV activity of various pine cone extracts increased with their acidity. Identification of the polyphenol groups as donors of the acidity was further supported by the observation that the anti-HSV activity of the natural polyphenolic products, such as tannin and lignin, significantly exceeded that of other natural or chemically modified antitumor polysaccharides. An experiment using radiolabeled virus particles indicated that the anti-HSV effect of both Fr. VI and lignin was attributable to interference with virus adsorption to these cells rather than to inhibition of virus penetration into the cells.

An in vitro reactivation system for investigation of herpes simplex virus latency.

An in vitro reactivation system for investigation of herpes simplex virus latency which is inexpensive, simple, and convenient to establish and operate is presented. The system is useful for investigation of compounds that may be involved in suppression or augmentation of herpes simplex virus reactivation. The uses of this model in exploring the mechanism of HSV reactivation, in screening for drugs which affect this process, and in testing their mode of action are discussed.

Synthesis of some phosphonates with antiherpetic activity.

Several keto phosphonates, phosphonoacetates , and dialkyl phosphonates containing (aryloxy)aryl groups were synthesized and evaluated for antiherpteic activity. Two of the most active compounds, 12 and 16, were evaluated topically in the mouse vaginal model against herpes simplex virus (HSV) type 2. Compound 16 exhibited an increased survival rate, as well as increased survival time. Evaluation of 16 in the guinea pig skin test against HSV-2 produced a reduction in virus titer, as well as in mean vesicle score.

The presence of RNA complementary to HSV-2 (herpes simplex virus) DNA in cervical intraepithelial neoplasia after laser therapy.

In-situ hybridization of radiolabelled DNA probes to tissue sections detected RNA complementary to HSV-2 (herpes simplex virus) DNA in 65% (22 of 34) biopsies from cervical intraepithelial neoplasia (CIN) and in none of internally paired benign epithelia from individuals before treatment by laser vaporization cone. Laser therapy did not produce an increase in clinical or subclinical HSV infections of the cervix. Of 18 patients with RNA complementary to HSV-2 DNA in CIN biopsies before treatment, in whom treatment was successful, virus transcripts were detected at follow-up in only two of them.

Effect of phosphonoformate on symptomatic genital herpes simplex virus type 2 infection of guinea pigs.

Using a guinea pig model of genital herpes simplex virus (HSV) type 2 infection, phosphonoformate treatment initiated after the onset of symptoms had a therapeutic effect on the outcome of disease. Severity of disease, viral shedding, and HSV-induced cellular changes in vaginal cytology were significantly reduced in drug-treated animals as compared to sham-treated animals, although virus was readily detected in the nervous system of animals from both groups. The percentage of animals harboring latent virus in their dorsal root ganglia was approximately the same in both drug-treated and sham-treated animals. No drug toxicity was found.

Failure of 2-deoxy-D-glucose in the treatment of experimental cutaneous and genital infections due to herpes simplex virus.

The effectiveness of 2-deoxy-D-glucose (2-DG) was evaluated in the treatment of cutaneous infections with herpes simplex virus type 1 (HSV-1) in mice and genital infections with HSV type 2 (HSV-2) in mice and guinea pigs. Groups of mice were inoculated in the lumbosacral or orofacial area with HSV-1 and treated topically three times a day with 0.2% or 0.5% 2-DG solution beginning 3 hr after inoculation. No effect on skin lesions, mortality, or latency was observed. Mice were inoculated intravaginally with HSV-2 and treated intravaginally three times a day with 0.2%-5% 2-DG in solution or suspended in miconazole nitrate cream beginning 6 hr, 24 hr, or 48 hr after inoculation. Replication of HSV-2 in the vagina and final mortality were not affected. Guinea pigs were infected intravaginally with HSV-2 and treated both intravaginally and topically on the external genitalia four times a day with 1% or 5% 2-DG in miconazole nitrate cream. Treatment initiated just prior to development of lesions (on day 3 after inoculation) did not alter vaginal virus replication, lesion development and severity, or virus titers in lesions.

Virus-infected colostral cell cytokine stimulation of human leukocyte natural killer cytotoxicity.

Natural killer cytotoxicity is an important antiviral defense mechanism. Human peripheral blood mononuclear cells cultured with herpes simplex virus (HSV)-infected cells produced a cytokine. This substance stimulated adult natural killer cytotoxicity from 53.0 +/- 10.5% to 79.8% (P less than 0.01) against HSV-infected target cells. These data resulted in a calculated cytokine-dependent cellular cytotoxicity (CDCC) value of 65.8%. Cytokine production was not stimulated by uninfected cells and was independent of the presence or absence of antibodies to HSV in sera of donors and mononuclear cells. Cells from human colostrum also produced an HSV-stimulated cytokine which mediated CDCC by using both adult (19.8 +/- 3.9%) and neonatal (18.6 +/- 3.4%) mononuclear effectors cells. Colostral cell cytokine production was also independent of donor HSV serology. Not all colostral cultures produced the cytokine, and in general colostrum-stimulated CDCC was lower than peripheral blood leukocyte-stimulated CDCC. Colostral cell cytokine stimulation of neonatal natural killer cytotoxicity may account in part for the increased nonspecific resistance of breast-fed infants to viral infection.

Anti-herpesvirus activity of adenine arabinoside analogues in tissue culture and a genital infection of mice and guinea pigs.

Four analogues of adenine arabinoside (ara-A) were compared for activity against herpes simplex virus (HSV) in tissue culture and in a genital infection of mice and guinea pigs. These analogues, 5'-monophosphate (ara-AMP), 5'-valerate ester (ara-AV), 2'3'-diacetate ester (ara-ADA), and 2',3',5'- triacetate ester (ara-ATA) have greater water and lipid solubility and resistance to deamination than ara-A. In mouse embryo fibroblast cells, similar viral inhibitory levels were noted with ara-A, AMP, and ara-Av, while ara-ADA and ara-ATA were 6-10 time less active. In mice infected intravaginally with HSV type 2 (HSV-2), intravaginal treatment with 10% concentrations of each of the compounds beginning 3 h after viral challenge, had no effect on infection rates, titers of virus in vaginal secretions, mortality rates or the mean day of death as compared with placebo-treated controls. In the HSV-2 genital infection of guinea pigs, treatment with 10% vaginal creams or placebo vehicle was initiated 6 or 24 h after viral inoculation. In animals treated at 6 h with ara-A, ara-AMP and ara-AV, there was complete inhibition of viral replication in the vaginal tract and development of external genital lesions. When treatment with these three drugs was delayed 24 h after infection, there was no effect on vaginal virus titers, but lesions severity was reduced by ara-A or ara-AMP therapy. Ara-ATA was ineffective whether begun at 6 or 24 h. The greater solubility in water and lipid as well as the resistance to deamination of ara-AMP and ara-AV did not appear to enhance their antiviral activity over that of ara-A. Additionally, ara-ADA and ara-ATA exhibited less activity both in tissue culture and in the experimental genital infections.

Fate of herpes simplex virus in lymphocytes from inflammatory joint effusions. i. Failure of the virus to grow in cultured lymphocytes.

The ability of lymphocytes isolated from the blood and synovial effusions of patients with rheumatoid arthritis to support the growth of herpex simplex virus (HSV) was studied. Whereas blood lymphocytes supported virus growth, effusion lymphocytes were usually non-permissive. Lymphocytes obtained from the effusions of patients with other forms of inflammatory arthritis or suspected viral arthritis commonly failed to support the growth of HSV, but the virus grew in effusion lymphocytes from patients with noninflammatory joint disease. Several nonspecific factors which might have accounted for this non-permissiveness were excluded.

Antiviral activity in the rabbit cornea of adenine arabinoside, Ara-A 5' monophosphate, and hypoxanthine arabinoside; and interactions with adenosine deaminase inhibitor.

The multiple microtrephination technique was used in the rabbit cornea to compare the activity against herpes simplex virus (hsv) of adenine arabinoside (ara-A), ara-A 5'monophosphate (ara-AMP) and hypoxanthine arabinoside (ara-Hx), and to determine the effect of addition of adenosine deaminase inhibitor (ADAI) to each. The greatest antiviral activity was shown by ara-AMP, and the least by ara-Hx. ADAI increased the activity of ara-A but had no effect on ara-AMP or ara-Hx.