To search the effective antisense oligonucleotide that inhibit the growth of Herpes simplex virus type 1 (HSV-1), six kinds of phosphorothioate oligodeoxynucleotides (S-ODNs) were synthesized and the antiviral activity was assessed by measuring cytopathic effect in Vero cells infected with HSV-1. Of the three dodecamer S-ODNs, cornplernentary to the translation initiation site of IE2 (AS2) and scrambled S-ODN (AS1) showed more significant antiherpetic activity than AS4 complementary to the IE4. Accordingly, the antiviral effect of dodecamer S-ODN was not specific. In contrast to the no inhibitory effect of sense strand S-ODN of ICP8 (AS6), two S-ODNs complementary to the translation initiation site of ICP8 (AS3) and that of IE1 (AS5) showed potent antiherpetic activity assessed in vitro HSV-1 virus yield assay. Antiherpetic effect of AS3 was decreased in proportion to the addition of AS6. The synthesis of viral protein ICPS and IE1 were inhibited in AS3 and AS5 treated HSV-1 infected Vero cells, respectively. These findings suggest that antiherpetic effect of AS3 is specifically mediated by targeting ICPS. S-ODNs had no effect on Vero cell viability. The data suggest that the 19-mer S-ODNs may be effective in antiviral chemotherapy.
Drug Therapy ; Herpes Simplex* ; Herpesvirus 1, Human ; Oligodeoxyribonucleotides ; Phosphorothioate Oligonucleotides* ; Simplexvirus* ; Vero Cells

Herpes simplex esophagitis has been recognized with increased frequency as an opportunistic infection in immunosuppressed or debilitated patients. However, it is also documented as self-limited esophagitis in apparently healthy patients. We report two cases of herpetic esophagitis diagnosed by biopsies. One case was noted in an immunocompetent patient having gastric peptic ulcer who had clinical improvement with symptomatic treatment. And the other was noted in an immunosuppressed patient having chemotherapy for gastric carcinoma who had resolution of symptoms with acyclovir therapy. Histologically, esophageal mucosa exhibited erosion and inflammatory cellular exudate with intranuclear eosinophilic inclusions in the epithelial cells. These were confirmed by the PCR and immunohistochemical stain for herpes simplex virus using a biopsy material.
Acyclovir ; Biopsy ; Drug Therapy ; Eosinophils ; Epithelial Cells ; Esophagitis* ; Exudates and Transudates ; Herpes Simplex* ; Humans ; Mucous Membrane ; Opportunistic Infections ; Peptic Ulcer ; Polymerase Chain Reaction ; Simplexvirus

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.
Amino Acid Sequence ; Base Sequence ; Catalytic Domain ; Chromatography, High Pressure Liquid ; Chromatography, Liquid ; Clone Cells ; DNA Packaging ; Drug Therapy ; Herpes Simplex* ; Herpesvirus 1, Human* ; Simplexvirus*

OBJECTIVE: JieZe-1 (JZ-1), a Chinese herbal prescription, has an obvious effect on genital herpes, which is mainly caused by herpes simplex virus type 2 (HSV-2). Our study aimed to address whether HSV-2 induces pyroptosis of VK2/E6E7 cells and to investigate the anti-HSV-2 activity of JZ-1 and the effect of JZ-1 on caspase-1-dependent pyroptosis. METHODS: HSV-2-infected VK2/E6E7 cells and culture supernate were harvested at different time points after the infection. Cells were co-treated with HSV-2 and penciclovir (0.078125 mg/mL) or caspase-1 inhibitor VX-765 (24 h pretreatment with 100 μmol/L) or JZ-1 (0.078125-50 mg/mL). Cell counting kit-8 assay and viral load analysis were used to evaluate the antiviral activity of JZ-1. Inflammasome activation and pyroptosis of VK2/E6E7 cells were analyzed using microscopy, Hoechst 33342/propidium iodide staining, lactate dehydrogenase release assay, gene and protein expression, co-immunoprecipitation, immunofluorescence, and enzyme-linked immunosorbent assay. RESULTS: HSV-2 induced pyroptosis of VK2/E6E7 cells, with the most significant increase observed 24 h after the infection. JZ-1 effectively inhibited HSV-2 (the 50% inhibitory concentration = 1.709 mg/mL), with the 6.25 mg/mL dose showing the highest efficacy (95.76%). JZ-1 (6.25 mg/mL) suppressed pyroptosis of VK2/E6E7 cells. It downregulated the inflammasome activation and pyroptosis via inhibiting the expression of nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing protein 3 (P < 0.001) and interferon-γ-inducible protein 16 (P < 0.001), and their interactions with apoptosis-associated speck-like protein containing a caspase recruitment domain, and reducing cleaved caspase-1 p20 (P < 0.01), gasdermin D-N (P < 0.01), interleukin (IL)-1β (P < 0.001), and IL-18 levels (P < 0.001). CONCLUSION JZ-1 exerts an excellent anti-HSV-2 effect in VK2/E6E7 cells, and it inhibits caspase-1-dependent pyroptosis induced by HSV-2 infection. These data enrich our understanding of the pathologic basis of HSV-2 infection and provide experimental evidence for the anti-HSV-2 activity of JZ-1. Please cite this article as: Liu T, Shao QQ, Wang WJ, Liu TL, Jin XM, Xu LJ, Huang GY, Chen Z. The Chinese herbal prescription JieZe-1 inhibits caspase-1-dependent pyroptosis induced by herpes simplex virus-2 infection in vitro. J Integr Med. 2023; 21(3): 277-288.
Caspase 1/metabolism* ; Inflammasomes/pharmacology* ; NLR Family, Pyrin Domain-Containing 3 Protein/metabolism* ; Pyroptosis ; Simplexvirus/metabolism* ; Drugs, Chinese Herbal/pharmacology* ; Herpes Simplex/drug therapy* ; Humans

Reports of combined candidal and herpetic esophagitis in immunocompetent states are rare and sporadic. A 44-year-old previously healthy lady presented with a one week history of progressive dysphagia, odynophagia and fever. Esophagogastroduodenoscopy (EGD) showed extensive desquamation of the entire esophagus except for distal 4 cm. Histopathological examination revealed ulcerated and inflamed squamous epithelium with the margin of ulcer showing a few overhanging squamous cells with dense eosinophilic cytoplasm, multinucleated and faceted nuclei with glassy chromatin, and an occasional Cowdry type A intranuclear inclusion bodies. Few candidal spores were seen in the underlying stroma. Intravenous acyclovir, fluconazole and pantoprazole were initiated. Oral analgesics were given for pain relief. She was treated for a total of 14 days. She showed significant improvement and was tolerating oral intake after discharge. The patient was asymptomatic with no evidence of recurrence at a 2-month follow-up.
Adult ; Antifungal Agents ; therapeutic use ; Antiviral Agents ; therapeutic use ; Candidiasis ; diagnosis ; drug therapy ; microbiology ; Esophagitis ; diagnosis ; drug therapy ; microbiology ; virology ; Esophagus ; microbiology ; pathology ; virology ; Female ; Herpes Simplex ; diagnosis ; drug therapy ; virology ; Humans ; Inclusion Bodies, Viral ; Spores, Fungal ; Treatment Outcome

OBJECTIVETo investigate in vitro and in vivo anti-human herpes simplex virus effects of photodynamic therapy (PDT) using 5-aminolevulinic acid (ALA).

METHODSGuinea pigs model of cutaneous herpes virus infection was applied, and Vero cells infected by HSV-I and HSV-II were used as experimental systems to observe the antiherpes effect of ALA-PDT.

RESULTSThe in vitro experiments showed that ALA-PDT has antiherpes effect on HSV-I and HSV-II, its effect was similar to that of acyclovir. The results of animal experiments showed that ALA-PDT had significant therapeutic effect on guinea pigs model of cutaneous herpes virus infection, the effect was dose-related.

CONCLUSIONALA-PDT could be effective in treating HSV infections, which may provide a new approach to the treatment of viral infections.

Aminolevulinic Acid ; therapeutic use ; Animals ; Cercopithecus aethiops ; Disease Models, Animal ; Female ; Guinea Pigs ; Herpes Simplex ; drug therapy ; virology ; Male ; Photochemotherapy ; Random Allocation ; Simplexvirus ; drug effects ; Skin Diseases, Viral ; drug therapy ; Treatment Outcome ; Vero Cells

OBJECTIVETrichosanthin (TCS), a ribosome-inactivating protein extracted from the root tuber of Chinese medicinal herb Trichosanthes kirilowii maximowicz, has various pharmacological properties including abortifacient, anti-tumor and anti-virus. This study aimed to evaluate the effects of TCS on infectious brain injury induced by Herpes simplex virus-1 (HSV-1) in mice.

METHODSNinety mice were randomly assigned into three groups: Normal control group (n=30), Model group (n=30) and TCS-treated group (n=30). Viral encephalitis was induced by intracranial inoculation of HSV-1 in the latter two groups. The TCS-treated group was injected with TCS 30 minutes before HSV-1 inoculation. The water content of brain tissue was measured at 1, 12, 24 and 48 hrs, and at 4 and 7 days after HSV-1 inoculation. The viral titer of brain tissue and brain histopathological changes were detected at 7 days after HSV-1 inoculation. The neurological deficient scores were determined daily.

RESULTSThe water content of brain tissue in the TCS-treated group between 48 hrs and 7 days after HSV-1 inoculation was significantly lower than that in the Model group (P < 0.05), although it was significantly higher than that in the Normal control group (P < 0.05). The viral titer of brain tissue in the TCS-treated group was markedly lower than that in the Model group (1.16 +/- 0.45 vs 2.89 +/- 0.44; P < 0.05) 7 days after HSV-1 inoculation. The neurological deficient scores of the TCS-treated group after 24 hrs of HSV-1 inoculation were significantly lower than that in the Model group but were higher than those of the Normal control group. TCS treatment resulted in alleviated pathological changes of brain tissue compared with the Model group 7 days after HSV-1 inoculation.

CONCLUSIONSTCS has protective effects against infectious brain injury induced by HSV-1 in mice.

Animals ; Body Water ; metabolism ; Brain ; metabolism ; pathology ; virology ; Encephalitis, Viral ; drug therapy ; Female ; Herpes Simplex ; drug therapy ; Herpesvirus 1, Human ; Male ; Mice ; Neuroprotective Agents ; therapeutic use ; Trichosanthin ; therapeutic use

PURPOSE: Gastric cancer is the most common malignancy in Korea. Although treatment such as surgery, chemotherapy, and immunotherapy has greatly improved, the mortality rate of gastic cancer is still high, A new therapeutic trial is necessary to improve the cure rate of gastric cancer. Therefore we investigated the pre-clinical significance of HSV-tk gene therapy using retroviral vector for gastric cancer cell lines. MATERIALS AND METHODS: LNC/HSV-tk retroviral vector and PA317/LNC/HSV-tk producer cell line were constructed. HSV-tk gene transduction and expression were detected by PCR. An in vitro ganciclovir(GCV) sensitivity test was performed by MTT assay. To evaluate in vivo GCV sensitivity, GCV was intraperitoneally injected after tumor formation in the nude mice. Bystander effect was observed in vitro MTT assay using YCC- S-2 cell line and in vivo using N87 and YCC-S-2 cell lines. RESULTS: The in vitro GCV sensitivity test showed that the growth inhibition was 30~32% with 0.5 uM GCV and 52~77% with 500 uM GCV in the HSV-tk transduced cell line in comparison with 0- 5% with 0.5 and 500 uM GCV in the parent cell line. The in vivo GCV administration showed that the tumors induced by HSV-tk transduced N87 cell line and YCC-S-2 cell line decreased completely, while the tumors with the parent cell lines continued to grow in nude mice. We observed no tumor cells in tissue specimen of the tumor induced by the N87/HSV-tk cell line after. GCV administration. In vitro and in vivo bystander effects were observed in HSV-tk/GCV system due to the resultant cell death exceeding the proportion of HSV-tk transduced cells in the mixtures of HSV-tk transduced and parent cells. CONCLUSION: HSV-tk transduced gastric cancer cell lines showed sensitivity to GCV and a bystander effect was observed. These results suggested that HSV-tk/GCV system should be evaluated in the clinical settings.
Animals ; Bystander Effect ; Cell Death ; Cell Line* ; Drug Therapy ; Ganciclovir* ; Genetic Therapy ; Herpes Simplex* ; Humans ; Immunotherapy ; Korea ; Mice ; Mice, Nude ; Mortality ; Parents ; Polymerase Chain Reaction ; Stomach Neoplasms* ; Thymidine Kinase* ; Thymidine* ; Zidovudine

Herpes simplex virus (HSV) is a recognized cause of gastrointestinal infection in immunodeficient patients. Although a few cases of HSV gastritis and colitis in immunocompromised patients have been reported, there are no reports of HSV duodenitis in patients with Crohn's disease (CD). A 74-year-old female was admitted with general weakness and refractory epigastric pain. She had been diagnosed with CD three years ago. Esophagogastroduodenoscopy (EGD) revealed diffuse edematous and whitish mucosa with multiple erosions in the duodenum. Considering the possibility of viral co-infection, cytomegalovirus (CMV) immunohistochemical staining, PCR, and cultures of duodenal biopsies were performed, all of which were negative with the exception of the isolation of HSV in culture. After administration of intravenous acyclovir for 1 week, follow-up EGD showed almost complete resolution of the lesions and the patient's symptoms improved. In CD patients with refractory gastrointestinal symptoms, HSV, as well as CMV, should be considered as a possible cause of infection, so that the diagnosis of viral infection is not delayed and the appropriate antiviral treatment can be initiated.
Acyclovir/therapeutic use ; Aged ; Antiviral Agents/therapeutic use ; Crohn Disease/complications/*diagnosis/virology ; DNA, Viral/analysis ; Duodenitis/complications/*diagnosis ; Endoscopy, Digestive System ; Female ; Herpes Simplex/*diagnosis/drug therapy/virology ; Humans ; Intestinal Mucosa/pathology ; Polymerase Chain Reaction ; Simplexvirus/genetics/*isolation & purification

In order to investigate the antiviral effect of chinonin against Herpes simplex virus (HSV), the encephalitis model in mice and skin infection model in guinea pigs were established by HSV- I and HSV-II infection respectively. Acyclovir was used as the positive reference drug to evaluate the antiviral capacity of chinonin. Chinonin showed an obvious therapeutic effect on encephalitis in mice at doses of 25 and 50 mg/kg. At both dosages, chinonin demonstrated stronger protection than acyclovir (1 and 5 mg/kg) to the infected mice from death. It was also found that chinonin could treat the skin infection in guinea pigs effectively. The therapeutic effect of chinonin was similar to that of acyclovir (5 mg/kg) at 25 mg/kg but obviously better than that at 50 and 75 mg/ kg. In conclusion, chinonin is a potential candidate for the treatment against HSV.
Acyclovir ; therapeutic use ; Animals ; Antiviral Agents ; pharmacology ; Encephalitis ; drug therapy ; virology ; Glycosides ; pharmacology ; Guinea Pigs ; Herpes Simplex ; drug therapy ; Herpesvirus 1, Human ; drug effects ; Herpesvirus 2, Human ; drug effects ; Mice ; Simplexvirus ; drug effects ; Skin Diseases, Infectious ; drug therapy ; Xanthenes ; pharmacology ; Xanthones