Human alphaherpesvirus 2 (HSV-2) is the main pathogen resulting human genital herpes, which poses a major threat to the socio-economic development, while there is no effective vaccine. In this study, we developed a novel lipopolyplex (LPP)-delivered mRNA vaccine expressing the HSV-2 envelope glycoprotein gD and evaluated its immunogenicity in mice. The mRNA vaccine was prepared from the genetically modified gD mRNA synthesized in vitro combined with the LPP delivery platform and it was named gD-ORI mRNA. The expression of gD antigen in the mRNA vaccine was validated in vitro by Western blotting and indirect immunofluorescence assay, then the immune responses induced by this mRNA vaccine in mice were evaluated. The immunization with gD mRNA alone induced strong humoral and cellular immune responses in mice. Robust and long-lasting gD-specific IgG antibodies were detected in the mouse serum after booster immunization with gD-ORI mRNA. The immunized mice exhibited a Th1/Th2 balanced IgG response and robust neutralizing antibodies against HSV-2, and a clear dose-response relationship was observed. The gD-specific IgG antibodies were maintained in mice for a long time, up to 18 weeks post-booster immunization. At the same time, multifunctional gD-specific CD4⁺ and CD8⁺ T cells in vaccinated mice were detected by intracellular cytokine staining (ICS). This novel gD-expressing mRNA vaccine delivered by LPP induces strong and long-lasting immune responses in mice post booster immunization and has a promising prospect for development and application. This study provides scientific evidence and reference for the development of a new mRNA vaccine for HSV-2.
Animals ; Herpesvirus 2, Human/genetics* ; Viral Envelope Proteins/genetics* ; Mice ; Herpes Genitalis/immunology* ; RNA, Messenger/immunology* ; Female ; Mice, Inbred BALB C ; Antibodies, Viral/blood* ; mRNA Vaccines/immunology* ; Antibodies, Neutralizing/blood* ; Humans

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

The study investigated the inhibitory effect and mechanism of tectorigenin derivative(SGY) against herpes simplex virus type Ⅰ(HSV-1) by in vitro experiments. The cytotoxicity of SGY and positive drug acyclovir(ACV) on African green monkey kidney(Vero) cells and mouse microglia(BV-2) cells was detected by cell counting kit-8(CCK-8) method, and the maximum non-toxic concentration and median toxic concentration(TC_(50)) of the drugs were calculated. After Vero cells were infected with HSV-1, the virulence was determined by cytopathologic effects(CPE) to calculate viral titers. The inhibitory effect of the tested drugs on HSV-1-induced cytopathy in Vero cells was measured, and their modes of action were initially explored by virus adsorption, replication and inactivation. The effects of the drugs on viral load of BV-2 cells 24 h after HSV-1 infection and the Toll-like receptor(TLR) mRNA expression were detected by real-time fluorescence quantitative PCR(RT-qPCR). The maximum non-toxic concentrations of SGY against Vero and BV-2 cells were 382.804 μg·mL~(-1) and 251.78 μg·mL~(-1), respectively, and TC_(50) was 1 749.98 μg·mL~(-1) and 2 977.50 μg·mL~(-1), respectively. In Vero cell model, the half maximal inhibitory concentration(IC_(50)) of SGY against HSV-1 was 54.49 μg·mL~(-1), and the selection index(SI) was 32.12, with the mode of action of significantly inhibiting replication and directly inactivating HSV-1. RT-qPCR results showed that SGY markedly reduced the viral load in cells. The virus model group had significantly increased relative expression of TLR2, TLR3 and tumor necrosis factor receptor-associated factor 3(TRAF3) and reduced relative expression of TLR9 as compared with normal group, and after SGY intervention, the expression of TLR2, TLR3 and TRAF3 was decreased to different degrees and that of TLR9 was enhanced. The expression of inflammatory factors inducible nitric oxide synthase(iNOS), tumor necrosis factor-α(TNF-α), and interleukin-1β(IL-1β) was remarkably increased in virus model group as compared with that in normal group, and the levels of these inflammatory factors dropped after SGY intervention. In conclusion, SGY significantly inhibited and directly inactivated HSV-1 in vitro. In addition, it modulated the expression of TLR2, TLR3 and TLR9 related pathways, and suppressed the increase of inflammatory factor levels.
Animals ; Antiviral Agents/therapeutic use* ; Chlorocebus aethiops ; Herpes Simplex/pathology* ; Herpesvirus 1, Human/metabolism* ; Isoflavones ; Mice ; TNF Receptor-Associated Factor 3/pharmacology* ; Toll-Like Receptor 2/metabolism* ; Toll-Like Receptor 3/metabolism* ; Toll-Like Receptor 9/metabolism* ; Toll-Like Receptors/metabolism* ; Tumor Necrosis Factor-alpha/metabolism* ; Vero Cells ; Virus Replication

In this study, emotional stress-induced herpes simplex virus type 1(HSV-1) susceptibility model was employed to simu-late the pathological state of " depression-induced liver fire", and the protection effect of Qingre Xiaoyanning(QX) in clearing liver fire was investigated. BALB/c mice were randomly divided into a normal group, a HSV-1 group, a restraint stress + HSV-1 group,low-(0. 658 g·kg~(-1)) and high-dose(1. 316 g·kg~(-1)) QX groups, and an acyclovir group. Except for the normal group and the HSV-1 group, the mice in other groups received daily restraint stress for 6 h from day 3 of medication. On day 9 of medication, mice were anesthetized by isoflurane and infected intranasally with HSV-1. Survival rate, weight change, encephalitis symptoms, and eye injury of mice were recorded for 14 d after virus infection. Hematoxylin-eosin(HE) staining and immunohistochemical staining were used to detect pathological changes and HSV-1 antigen distribution. Plaque assay was performed to detect the titer of HSV-1. The protein ex-pression of ICP27 in the mouse brain was detected by Western blot. The experimental results showed that QX could increase the survival rate of HSV-1-infected mice loaded with emotional stress(P<0. 001), reduce the titer of HSV-1 in the mouse brain(P<0. 01), relieve brain inflammation(P<0. 05) and eye injury(P<0. 05), down-regulate the expression of ICP27 related to HSV-1(P<0. 05), and decrease the distribution of HSV-1 antigen in the mouse brain. The results demonstrated that QX significantly reduced the susceptibility to HSV-1 induced by emotional stress, which is expected to provide a theoretical basis for the treatment and preven-tion of HSV-1 infection and promote the clinical development and application of Chinese medicine effective in clearing liver fire.
Animals ; Capsules ; Herpes Simplex ; Herpesvirus 1, Human ; Mice ; Mice, Inbred BALB C ; Psychological Distress

ABSTRACT The symptoms of recurrent oral herpes infection may vary, from mild discomfort to life threatening. Dentists are more likely to be consulted in this oral infection, hence the ability to diagnose and treat this disease is mandatory. This article described manifestation and therapy of recurrent oral herpes infection. In the first case, a 41-year-old woman came with a complaint of painful sore mouth in her lower gingiva. She experienced canker sores for two to three times a year on the tongue, lips and palate. Laboratory results showed positive IgM anti HSV-2, which matched with the primary herpes infection. However, based on the history and clinical manifestations, she was diagnosed with recurrent intra oral herpes infection. In the second case, a 70-year-old man came with a very painful canker sores in his entire mouth. The patient had to be hospitalised for three days and received analgesic, antibiotic and gel containing triamcinolone. After hospitalisation, canker sores did not heal. Extraorally, we found a crust in the vermillion border of the upper lip and intraorally, we found multiple ulceration of keratinised and non-keratinised mucosa. The use of topical steroid in this patient may aggravated ulceration due to its ability to cause rapid spreading of the virus. The first patient was given chlorhexidine gluconate 0.2% and the second patient was prescribed with doxycycline rinse. Both patients received multivitamin containing zinc. Complete history taking, objective and adjunctive examination played a role in establishing the diagnosis and treatment of recurrent oral herpes infections.
Stomatitis, Herpetic

PURPOSE: To report a case of a Corynebacterium macginleyi-infected corneal ulcer of a patient who had been treated for conjunctivitis for more than 3 months. CASE SUMMARY: A 72-year-old female was transferred from a private ophthalmic clinic for evaluation of herpetic keratitis with progressive corneal edema and infiltration in the left eye. She had a history of conjunctival hyperemia and eyeball pain in her left eye 3 months prior to her visit. She was treated with levofloxacin eye drops and acyclovir ointment (Herpesid®, Samil, Co., Ltd. Seoul, Korea). On slit lamp examination, 5.4 × 4.0 mm corneal epithelial defects and stromal infiltrations were observed in the upper to central cornea, and endothelial keratic precipitates were found. Gram positive bacteria were detected on Gram staining and Corynebacterium macginleyi was identified on bacterial cultures from the conjunctiva and cornea. She was treated with topical vancomycin eye drops. After 3 months of treatment, the corneal ulcer was completely resolved, leaving mild superficial opacity on the cornea. CONCLUSIONS: While Corynebacterium macginleyi, normal flora of the conjunctiva, is considered a major causative agent for conjunctivitis and blepharitis, Corynebacterium macginleyi should also be considered a possible cause of slowly progressive keratitis in patients with chronic conjunctivitis.
Acyclovir ; Aged ; Blepharitis ; Conjunctiva ; Conjunctivitis ; Cornea ; Corneal Edema ; Corneal Ulcer ; Corynebacterium ; Female ; Gram-Positive Bacteria ; Humans ; Hyperemia ; Keratitis ; Keratitis, Herpetic ; Levofloxacin ; Ophthalmic Solutions ; Seoul ; Slit Lamp ; Vancomycin

PURPOSE: We report a case of herpes simplex keratitis after Descemet membrane endothelial keratoplasty (DMEK). CASE SUMMARY: A 67-year-old male underwent DMEK in his left eye due to pseudophakic bullous keratopathy. One week after DMEK, re-bubbling was performed due to partial detachment of Descemet's membrane at the corneal periphery. After re-bubbling, the cornea remained clear and the patient's visual acuity gradually improved. Two months after DMEK, the patient presented with mild discomfort and decreased visual acuity. The cornea showed an irregular, narrow dendrite with an epithelial defect and surrounding opacity. After confirming that Descemet's membrane was attached, the patient was started on oral valacyclovir for suspected herpes keratitis. Herpes simplex virus type 1 was eventually identified by polymerase chain reaction. The corneal lesion resolved after three weeks of antiviral treatment. CONCLUSIONS: Similar to penetrating keratoplasty, DMEK can trigger outbreaks of herpes simplex keratitis. Herpes simplex keratitis should remain on the clinician's differential diagnosis for patients who present with a corneal epithelial irregularity and decreased visual acuity following DMEK.
Aged ; Cornea ; Corneal Transplantation ; Dendrites ; Descemet Membrane ; Diagnosis, Differential ; Disease Outbreaks ; Herpes Simplex ; Herpesvirus 1, Human ; Humans ; Keratitis ; Keratitis, Herpetic ; Keratoplasty, Penetrating ; Male ; Polymerase Chain Reaction ; Visual Acuity

Chest pain in kidney transplant patients is usually caused by cardiac or pulmonary problems. However, it may be rarely caused by opportunistic esophageal infections. A 66-year-old female kidney transplant recipient was admitted because of chest pain. She had been treated with high-dose steroid and immunosuppressants for acute T-cell-mediated rejection. Cardiologic and pulmonary evaluations had normal results. Endoscopic examination revealed three clear ulcerative lesions in the esophagus. Histological and immunohistochemical staining of the endoscopic biopsy specimens revealed coinfection of herpes simplex virus and cytomegalovirus. The patient was treated with intravenous ganciclovir for 2 weeks. Her symptoms completely resolved, and follow-up endoscopy revealed complete healing of the previous ulcers. Viral esophagitis should be considered in the differential diagnosis in kidney transplant recipients presenting with chest pain.
Aged ; Biopsy ; Chest Pain ; Coinfection ; Cytomegalovirus ; Diagnosis, Differential ; Endoscopy ; Esophagitis ; Esophagus ; Female ; Follow-Up Studies ; Ganciclovir ; Herpes Simplex ; Humans ; Immunosuppressive Agents ; Kidney ; Kidney Transplantation ; Simplexvirus ; Thorax ; Transplant Recipients ; Ulcer

BACKGROUND: Kaposi varicelliform eruption (KVE) is a disseminated viral infection primarily caused by the herpes simplex virus in the setting of an underlying chronic skin disease. Few studies have reported the clinical characteristics and predisposing factors for recurrent KVE.OBJECTIVES: To characterize the clinical features and predisposing factors for recurrent KVE.METHODS: This retrospective comparative study of recurrent vs. single-episode KVE was performed at the Pusan National University Hospital between 2004 and 2017.RESULTS: A total of 84 episodes occurred in 60 patients, and of these, 13 patients developed recurrence (21.7%). No statistically significant intergroup difference was observed in the mean age and sex distribution. The face was the most common site of involvement in both groups, followed by the trunk and the upper and lower extremities. Atopic dermatitis was the most common pre-existing disease in both groups; however, Darier's disease was more common in the recurrent KVE group, and this difference was statistically significant. Most patients with KVE (66.7%) showed aggravation of the underlying skin disease within 3 months of KVE onset. This finding was more prominent in patients with recurrent episodes (91.7%) than in those with single-episode KVE (58.3%), (p=0.040).CONCLUSION: This study can contribute to a better understanding of recurrent KVE and guide clinicians in treating patients with conditions predisposing to KVE.
Busan ; Causality ; Darier Disease ; Dermatitis, Atopic ; Humans ; Kaposi Varicelliform Eruption ; Lower Extremity ; Preexisting Condition Coverage ; Recurrence ; Retrospective Studies ; Sex Distribution ; Simplexvirus ; Skin Diseases

Disseminated neonatal herpes simplex virus (HSV) infection is a severe disease with a high mortality rate. Here, we report the patient presented with fulminant hepatic failure secondary to HSV infection followed by renal failure without any mucocutaneous symptoms. The patient recovered after treatment with acyclovir and peritoneal dialysis. This is the first known case of a patient in Korea who survived disseminated HSV infection with fulminant liver failure followed by renal failure without undergoing liver transplantation.
Acyclovir ; Hepatitis ; Herpes Simplex ; Humans ; Infant, Newborn ; Korea ; Liver Failure ; Liver Failure, Acute ; Liver Transplantation ; Mortality ; Peritoneal Dialysis ; Renal Insufficiency ; Simplexvirus