Effect of Preincubation of Thymopentin and Thymopentin-Treated Mononuclear Cell Supernatant on Herpes Simplex Virus Infection of Human Fibroblast Monolayers

Kim, Hyung Ok,Lee, Jeong Deuk,Kim, Tae Yoon,Kim, Chung Won CATHOLIC MEDICAL CENTER 1993 Bulletin of the Clinical Research Institute Vol.21 No.1

Herpes simplex viruses are one of the most common sources of infectious in man. Relapses occur frequently in a small fraction of the individuals who had a primary infection. Many different modes of treatment have been tried for herpes simplex. Thymopentin is a suggestive treatment agent for herpes simplex virus infection. In the previous studies, thymopentin induced changes in cell-mediated immunity. However, it has not been investigated to the direct effect of thymopentin on herpes simplex virus infection. In this study, we studied the direct effect of thymopentin on herpes simplex virus infection of human fibroblast monolayers. Mononuclear cells were obtained from herpes simplex patients and healthy adults. The cells were cultured in the presence of different thymopentin concentrations (10^5, 10^4, 10^2, 10^1, 10^0, 10^-1, 0 ng/dl). The supernatant was harvested. The fibroblast monolayers were incubated in the presence of different thymopentin concentrations (10^5, 10^4, 10^2, 10^1, 10^0, 10^-1, 0 ng/dl) and supernatant for 30 minutes. The fibroblast monolayers were inoculated by herpes simplex virus. The number of herpes simplex virus infected cells was determined by plaque assay and immunofluorescence. The results were as follows: 1. The maximum cytopathic effect on the infected fibroblast monolayers was observed for 48-72 hours after infection. Immunofluorescence of the infected fibroblast was preceded than the cytopathic effect appeare4. 2. Thymopentin increased plaque forming unit (PFU) to 43.4% at 10^3 ng/ml, 33.0% at 10&1 ng/ml and immunofluorescence to 2.93%, 2.67% respectively. 3. Thymopentin in mononuclear tell supernatant from herpes simplex patients decreased PFU to 12.3% and immunofluorescence to 3.68% at 10^3 ng/ml. 4. Thymopentin in mononuclear cell supernatant from healthy adults decreased PFU to 5.7% at 10^5, l0^3 and 10^2 ng/ml thymopentin. In conclusion, thymopentin directly increased herpes simplex virus infection in cultured fibroblast monolayers at 10^3 ng/ml (p<0.05).

방사선 조사가 단순포진 바이러스의 증식에 미치는 영향

이유철,조유진,신상훈 대한감염학회 1999 감염 Vol.31 No.2

Herpes Simplex Virus Type-1 (HSV-1) 감염에 따른 세포내 유리 $Ca^{2+}$농도의 변화

남윤정,이규철,이찬희 한국미생물학회 2000 미생물학회지 Vol.36 No.4

Herpes simplex virus type-1 (HSV-1)의 감염에 따른 세포내 유리 칼슘농도의 변화에 대한 실험을 수행한 결과, HSV-1이 Vero 세포에 감염한 후 4시간째에 세포내 칼슘농도가 최대로 감소한 것을 알았으며 이러한 세포내 유리 칼슘농도의 감소는 감염성 바이러스의 양에 따라 커지며, 유전자 발현 억제제의 처리나 바이러스의 불활성화에 의해 극복되었다. 따라서 바이러스의 유전자발현이 세포내 유리 칼슘농도의 감소에 중요한 역할을 한다는 것을 알 수 있다. 또한 Vero 세포에 바이러스를 감염시키고 미세소관 안정제인 taxol을 처리하여 4 시간째의 세포내 유리 칼슘농도의 감소가 극복된다는 사실로부터 바이러스이 유전자 물질의 이동에는 미세소관이 관여한다는 것을 알 수 있었다. 이와 같은 실험 결과로부터 Vero 세포에서 HSV-1에 의해 유도되는 세포내 유리칼슘 농도의 감소는 HSV-1 증식과 밀접한 관계를 가진다고 생각된다. 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.

잠재성 Herpes Simplex Virus의 재활성화에 대한 n-Butyrate의 효과

천연숙(Yeon-Sook Chun),박노희(No-Hee Park) 대한약리학회 1986 대한약리학잡지 Vol.22 No.2

4-carbon fatty acid인 n-Butyrate(n-BTA)는 herpes virus의 일종인 Epstein-Bat virus(EBV)에 작용해서 잠복형인 EBV를 활성형태로 유도시키는 것으로 알려져 있다. 고로 본 실험에서는 mouse의 삼차신경절에 잠복하고 있는 HSV의 재활성화에 대한 n-BTA의 효과를 실험 관찰하였다. Pentobarbital로 마취시킨 mouse의 양쪽눈 각막을 30 gauge 주사바늘로 scarify한 후에 type I HSV(HSV-1) 10μ1(1 X 10⁵ plaque-forming units)를 각각 점안 감염시켰다. virus를 감염시킨 4주 후에 mouse의 삼차신경절을 적출하여 시험관 내에서 조직배양을 시행하였다. 조직배양시에 0. 1, 0. 25, 0.5. 1.0 그리고 2.0mM농도의 n-BTA를 첨가하였으며 1일, 2일, 3일간 각각 배양한 후 신경절을 연마하여 연마액내의 HSV-1 titer를 Vero cell monolayer에서 plaque assay로 측정하였다. 1) n-BTA첨가군은 잠재성 HSV가 대조군에 비하여 현저하게 빨리 재활성화 되었고 재활성화되는 virus의 양도 현저히 증가되었다. 2) 24시간을 계속해서 n-BTA 각 농도를 첨가해서 배양할 군은 n-BTA 6시간 첨가 배양하고 새로운 배양액으로 갈아서 18시간 배양한 군에 비해 잠재성 virus의 재활성화가 현저히 증가되었다. 3) Gang1ionic latent HSV-1의 재 활성화에 영향을 미치는 각 농도의 n-BTA는 Vero cell의 monolayer에서의 HSV-1의 번식에는 아무런 영향을 미치지 않았다. 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.

Epidemiology of Herpes Simplex Virus Type 1 and Herpes Simplex Virus Type 2 Infections: A Single-Center Retrospective Study in Korea

( Dong Seok Shin ),( Joo Yeon Ko ) 대한피부과학회 2021 대한피부과학회지 Vol.59 No.6

Background: Herpes simplex virus (HSV) infection is a common viral disease of the skin worldwide. HSV type 1 (HSV-1) has been associated with orofacial infections, while HSV type 2 (HSV-2) accounts for the main cause of genital herpes. However, a number of recent clinical studies have revealed that the epidemiology of HSV-1 and HSV-2 is changing. Objective: We performed the study to find out if there is any correlation between HSV subtypes and other multiple variables. Methods: We examined the results of HSV-polymerase chain reaction (PCR) tests performed in our hospital from July 2017 through June 2020. We analyzed the HSV subtypes of samples from 304 patients that showed positive HSV-PCR. Comparative analysis was performed to identify the correlation between HSV subtypes and other multiple variables. Results: HSV-1 was typed in 53.9% (164/304) and HSV-2 in 46.1% (140/304) of the patients during the study period. HSV-1 showed predilection for body area above the waist, whereas HSV-2 was dominant below the waist. There was a statistically significant correlation between the HSV subtypes and site of infection (p<0.001). Female patients were more likely to acquire HSV-2 infection than male patients (p=0.027). The ratio of HSV-2 infection tended to get bigger as the age of patients increased (p<0.001). There was not a significant difference in HSV subtypes among the groups defined according to immune status (p=0.333). Conclusion: Our study provided abundant data concerning various aspects of epidemiology of HSV infection. They can be used to plan and evaluate strategies for management of patients. (Korean J Dermatol 2021;59(6):462∼467)

Epidemiology of herpes simplex virus type 1 and type 2 infections in Korea: a retrospective singlecenter study

( Na Young Kim ),( Dong Seok Shin ),( Young Wook Ko ),( Dong Uk Cheon ),( Won Seon Koh ),( Jeong Eun Kim ),( Young Suck Ro ),( Joo Yeon Ko ) 대한피부과학회 2019 대한피부과학회 학술발표대회집 Vol.71 No.2

Background: Herpes simplex virus (HSV) type 1 is main cause of orofacial infection, whereas HSV-2 is usually associated with genital infection. HSV-2 tends to have more frequent recurrence and severe symptoms compared to HSV-1. Recent studies reported the incidence of HSV-1 infection has been increasing up to about 50% in genital area in United States. Objectives: The aim of this study was to estimate the proportion of HSV-1 and HSV-2 according to various factors such as infection site, age, sex, and underlying disease in Korea. Methods: We performed a retrospective study of 220 Korean patients diagnosed as recurrent HSV infections by HSV PCR skin testing from 2016 to 2018. The prevalence of infection with HSV-1 and HSV-2 were compared according to the site, sex and age. Results: Of the 220 participants, 124 became infected with HSV-1 and 96 with HSV-2. HSV-1 was main cause of orofacial herpes (95.97%, p<0.001) and HSV-2 showed 88.54% of genital herpes. Female was more prevalent in both HSV-1 and HSV-2 infections than male, however HSV-2 infection showed significant female dominance (p<0.02). Significant increasing trends of HSV-2 infections by age in comparison with HSV-1 have been revealed (P=0.005). Conclusion: HSV-2 was still major cause of genital herpes in this study. Further large-scale Korean epidemiologic studies are needed because the recurrence of HSV infection has negative effect on patients’ quality of life.

한약단미제 및 탕제의 항 Herpes Simplex Virus Type-1 활성탐색

강봉주,양기상,김명희,박갑주,Kang, Bong-Joo,Yang, Ki-Sang,Kim, Myung-Hee,Park, Kap-Joo 대한미생물학회 1997 Journal of Bacteriology and Virology Vol.27 No.2

향약집성방 및 동의보감등 한의학 서적을 기초로 하여 선택한 탕제 45종과 단미제 80종의 메탄올과 열수추출물을 가지고 MTT assay를 실시하여 항Herpes simplex virus-1 (HSV-1)에 대한 활성을 조사하였다. 열수추출 탕제 45종 중 백사전방, 현람방I, 현람방II, 반천청방 4종에서 항 HSV-1 활성을 나타냈고, 이들의 SI (selective index)값은 $2.1{\pm}0.5$에서 $11.8{\pm}2.2$ 범위의 값을 가졌으며 메탄올추출 탕제 45종 중에서는 단치시호탕III, 반천청방, 정향울금방, 대황오배자고, 홍인락삼등방, 호장해독탕에서 활성을 보였고 이들의 SI값은 $1.7{\pm}0.2$에서 $10.5{\pm}3.1$ 범위의 값을 나타냈다. 열수추출 단미제 중계지, 관중, 구인분, 대황, 자화지정, 포공영, 호장근, 황백등 8종에서 항 HSV-1 활성이 나타났고, 이들의 SI값은 $1.6{\pm}0.1$에서 $10.2{\pm}0.7$ 범위의 값을 가졌으며 메탄올추출 탕제 중에서는 계지, 목방기, 상지, 호장근에서 활성을 보였고 이들의 SI 값은 $2.9{\pm}1.5$에서 $9.3{\pm}0.5$ 범위의 값을 나타내었다. 처방제 중 반천청방은 열수추출물과 메탄올 추출물 모두에서 항바이러스 활성을 나타냈고, 단미제 중 계지와 호장근도 열수와 메탄올 추출물 모두에서 항바이러스 활성을 보였다. 이들 탕제 및 단미제는 분획 및 분석실험을 실시하여 활성성분을 추적하고 있다. In order to search for anti-Herpes simplex virus (HSV) type-1 agents from Korean medicinal herbs and Korean traditional prescriptions (herb complexes), we selected 80 medicinal herbs and 45 prescriptions, based on a review of the Korean traditional medicine books. Both methanol extracts and boiling-water extracts were tested by means of the MTT assay (tetrazolium based colorimetric assay). Ten of the 125 methanol extracts: CM-11, CM-18, CM-19, CM-21, CM-22, CM-39, MM-3, MM-18, MM-29, MM-73 (see explanation of nomenclature below), showed efficacy against HSV-1. Twelve of the water extracts: CW-2, CW-3-I, CW-3-II, CW-18, MW-3, MW-5 MW-6, MW-12, MW-47, MW-69, MW-73 and MW-79 were active. #3 (individual herb) and #73 (individual herb) were interesting because both water and methanol extracts were active. Especially, #3 is a part of composition of Hong-il-$laksamd{\check{u}}ngbang$ and Hojanghaedokt'ang which have anti-HSV-1 activitives. The SI value of MW-69 and CW-18 was relative high as $10.2{\pm}0.7$ and $11.8{\pm}2.2$. The cytotoxic effect on Vera cells of $Panch'{\check{o}}nch'onch'{\check{o}}ngbang$, Taraxacum platycarpum H. Dahlst. and acycloguanosine was determined by MTT assay. Water extracts of $Panch'{\check{o}}nch'onch'{\check{o}}ngbang$ (prescription) and Taraxacum platycarpum H. Dahlst. showed very weak cytotoxic effects on Vero cells at > $100\;{\mu}g/ml$ but acycloguanosine showed strang cytotoxic effects on Vera cells at > $100\;{\mu}g/ml$. As a result, #3, #73, MW-69 and CW-18 are considered as potentially useful for anti-HSV-1 agent and will be the focus of further research. Abbreviations: CM - methanol extracts of traditional prescriptions; CW - water extracts of traditional prescriptions; MM - methanol extracts of individual herbs; MW - water extracts of individual herbs.

토양으로부터 HIV 바이러스에 항바이러스 활성을 나타내는 미생물의 탐색

김성욱,이상한,정태숙,문석식,이종교,이항우,이원영,복성해 한국산업미생물학회 1993 한국미생물·생명공학회지 Vol.21 No.5

토양으로부터 분리한 520여 각종 미생물을 대상으로 항 바이러스 활성을 조사한 결과 HSV-1 바이러스에 95% 이상의 저해 활성을 나타낸 것이 6.9%이었으며, 이 시료들은 HSV-2 바이러스에서도 62.1%가 95% 이상의 활성을 나타내었다. 이들중 HIV 항원의 발현이 양성이고 80% 이상의 역전사 효소 활성 저해를 나타내는 2개의 시료를 대상으로 HIV 바이러스에 대한 저해 활성을 조사하여 HIV 바이러스에 강한 저해 활성을 나타내는 8S-301 시료를 선별하였다. For screening of the antiviral agent from soil, about 520 strains of microorganisms were evaluated for their antiviral activity. About 6.9% of strains showed more than 95% antiviral activity against Herpes Simples Virus (HSV)-1. Two strains among 30 strains active against HSV-1 virus showed a quite strong activity against human immunodeficiency virus.

전자현미경 기법을 이용한 Herpes simplex 2형 바이러스 항원의 면역학적 분석

김천식,오명환 한국미생물학회 2004 미생물학회지 Vol.40 No.1

Antigenic analysis of Herpes simplex type 2 virus was performed and its major antigen was localized using an immunoelectron microscopy. Antigens of 32, 43, 59 and 69 kDa were constantly expressed during the course of infection for 48 hr in the infected Vero cell. An antigen of 51 kDa was turned out to be the major one in inducing a immune response in Western-blot analysis. The 51 kDa antigen was localized on the surface of HSV-2 by immunoelectron microscopy using colloidal golds and anti-HSV 2 polyc1onal antibody. Immunofluorescence assay indicated that viral antigens were found throughout the infected cell and, especially, on the surface of the cell. 단순포진 바이러스 감염을 유발하는Herpes simplex2형 바이러스의 감염에 관여하는 항원들과 중화항체 생산을 유발하는 주요 항원들의 위치를 확인하였다. Vero cell에 감염하였을 때 48시간 동안 31, 43, 59, 69 kDa 바이러스 항원들이 지속적으로 발현되었으며, 감염된 쥐에서 생산한 항체와의 반응에서는 51 kDa 항원이 가장 강한 반응을 보였다. 면역전자현미경으로 위치를 확인한 결과 colloidal gold가 바이러스 표면에 발견되는 것으로 보아 이 항원이 바이러스 표면에 존재하고 있는 것으로 확인되었다. 형광현미경 분석은 이 항원들이 감염된 세포 내에서 전반적으로 발견되었고 특히 세포 표면에서 많이 발현되고 있었다.

항바이러스 효과에 응용되는 한약의 연구 동향

임성우,Lim, Sung-Woo 대한암한의학회 1998 大韓癌韓醫學會誌 Vol.4 No.1

In order to find antiviral effect against Human immunodeficiency virus(HIV), Herpes simplex virus type I(HSV-1) and II(HSV-2) from herb medicines, publicated 29 paters on anti-viral effect of herbal medicines and a convenient virus-induced cytopathic effect (CEP) inhibition assay was introduced. The major virus on experiment are HIV, Hepatitis B virus and HSV-1,2. Those of other studies showed inhibition of infected virus DNA replication and screening test of herbal medicines. More than 15 extractions were prepared by pure water boiling from herbal medicines, and their toxicity of infected cell and anti-viral activities were evaluated. Among them, the major part of herbal medicines showed cell stability compared with the contrast. Cytotoxic concentration (CC) of the $H_2O$ extracts of Padoo against HIV was <4.0, Hyungbangpaedoksan against HIV was 9.3, Whangyonhaedoktang against HIV-1 and HSV-2 was 15.3. These are high level cytotoxic concentration compared with the contrast. But antiviral effect was unable to figure out for selective $index(SI)=CC_{50}/EC_{50}$. The other herbal medicines were unable to showed potent anti-HIV and anti-HSV activity. The antiviral activation using herbs in this thesis have unlimited objects, to select research object will help to show the direction of antiviral drug development that have less side effect and more excellent efficiency.