• 미생물학회지
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• 제36권4호
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• pp.306-311
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• 2000

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.

• 한국한의학연구원논문집
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• 제1권1호
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• pp.477-493
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• 1995

In order to facilitate the molecular characterization of the Herpes simplex Virus types 1 and types 2 genome DNAs, a gene library of cloned restriction frtgments have been produced. The Vero cells were infected with HSV-1 and HSV-2. 48 hours after infection, the infected cells Ivere Iysed, and multinucleated giant cells were observed approximately at seventy-two hours postinfection. The multiplication of HSV-1 and HSV-2 was observed in Vero cells using electromicroscopy. The nucleocapsids in nuclei were obseryed, and the assembled virions were budded out through the vacuole, and the virions were released from the cells. HSV-1 and HSV-2 was analyzed by digestion of their genome DANs with restriction ensymes. HSV-1 and HSV-2 genome DNAs were digested with BarnHI, Bgfl respectively. The BarnHI rlestriction fragments of HSV-1 and HSV-2 genome DNAs were twenty-seven fragments and thair molecular sizes were ranging $0.70{\sim}15.08$, $4.4{\sim}31.0$ tilobases. The BglII restriction fragments of HSV-1 and HSV-2 genome DNAs were sixteen, eighteen fragments and thair molecular sizes were ranging $4.8{\sim}30.0$, $1.2{\sim}25.0$ kilobases. And then BglII restriction frgments were cloned in Escherichia coli(E.coil) using the plasmid vector pBacPAK9.

• 한국미생물·생명공학회지
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• 제29권1호
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• pp.62-66
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• 2001

This study was performed to investigate antigenic expression patterns in the course of HSV-1 infection. In SDS-PAGE analysis, HSV-1 antigens were detected, and among them, antigens in the size of 39, 47, 63, 86, 101, 105, 135, 159, and 181 kDa appear to be expressed in the most dominant forms. BALB/c mice were infected with HSV-1 for 29 days and antigenic expression from HSV-1 was investigated by Western blot analysis using anti-HSV-1 sera collected every two days from BALB/c mice infected with HSV-1. Most of HSV-1 antigens appeared sporadically as the infection progressed. However, antigens in the sizes of 63kDa and 135kDa were expressed from day 1 and 3, respectively, and existed continuously during the course of infection for 29 days, suggesting that they are the most dominant antigens inducing immune response durign HSV-1 infection, and they could be the target antigens for the development of vaccines. The isotype levels of IgA, IgGl, and IgM increased till the 17 th day infection and then started to decrease. During this course. IgGl was the most dominant isotype. In an indirect immunofluorescent assay, antibodies exhibited surface binding to the Vero cell infected with HSV-1, demonstrating that HSV-1 antigens are expressed on the surface of Vero cells.

• 대한핵의학회지
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• 제38권1호
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• pp.62-73
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• 2004

목적: 단순 헤르페스 제 1형 티미딘 키나제(herpes simplex virus type 1 thymidine kinase gene: HSV1-tk)는 GCV와 함께 유전자치료의 한 방법으로서 가장 활발하게 연구되어왔으며, HSV1-tk 효소에 대한 다양한 기질들이 연구되어서 이를 보고 기질로 한 비침습적인 HSV1-tk 유전자 영상시스템에서 가장 널리 사용되고 있다. 본 연구에서는 보고기질로서 방사성 요오드가 표지된 5-iodovinyl-2-deoxyuridine (IVDU) and 5 -iodovinyl-2-fluoro-2-deoxyuridine (IVFRU)를 보고 기질로 하여 HSV1-tk 유전자 영상시스템에서의 유용성을 확인하고자 하였다. 대상 및 방법 HSV1-tk 유전자 영상을 위하여 HSV1-tk 유전자를 레트로 바이러스 벡터를 이용하여 Morris hepatoma 세포주(MCA-tk)에 이입한 세포주를 제조하였으며, HSV1-tk 유전자의 발현을 확인하기 위하여 Northern blotting과 Western Blotting을 시행하였다. 대조세포주인 MCA와 제조된 MCA-tk 세포주에 방사표지 IVDU와 IVFRU를 이용하여 480분까지 세포내 섭취율을 평가하였으며, 또한 MCA-tk 세포주의 백분율을 증가시키면서 이에 따른 IVDU와 IVFRU의 섭취율을 평가함으로서 섭취율과 세포수와의 상관관계를 평가하였다. 결과: MCA-tk 세포주에서 HSV1-tk의 mRNA의 발현과 HSV1-TK 단백질의 발현을 확인하였다. 방사성 요오드 표지 IVDU와 IVFRU 모두는 MCA 세포주에서는 아주 낮은 섭취율을 나타내었으며, MCA-tk 세포주에서는 모두 증가된 섭취를 보였다. IVDU가 480분에서 IVFRU보다 4배 높은 섭취를 나타내었다(p<0.01). 방사표지 IVDU와 IVFRU 모두에서 MCA-tk의 백분율의 증가에 따라서 직선적인 상관관계($R^2>0.96$)를 나타내었다. 결론: 방사성 요오드 표지 IVDU와 IVFRU는 HSV1-tk유전자가 이입된 간암세포주에서 모두 특이적인 높은 섭취율을 나타내고 직선적인 상관관계가 나타나서 두 기질 모두 HSV1-tk유전자 영상시스템에서 보고 기질로서 유용하게 사용될 수 있을것으로 기대된다.

• Journal of Ginseng Research
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• 제48권4호
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• pp.384-394
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• 2024

Background: Herpes simplex virus type 1 (HSV-1), known to latently infect the host's trigeminal ganglion, can lead to severe herpes encephalitis or asymptomatic infection, potentially contributing to neurodegenerative diseases like Alzheimer's. The virus generates reactive oxygen species (ROS) that significantly impact viral replication and induce chronic inflammation through NF-κB activation. Nuclear factor E2-related factor 2 (Nrf2), an oxidative stress regulator, can prevent and treat HSV-1 infection by activating the passive defense response in the early stages of infection. Methods and results: Our study investigated the antiviral effects of ginsenoside Rg5, an Nrf2 activator, on HSV-1 replication and several host cell signaling pathways. We found that HSV-1 infection inhibited Nrf2 activity in host cells, induced ROS/NF-κB signaling, and triggered inflammatory cytokines. However, treatment with ginsenoside Rg5 inhibited ROS/NF-κB signaling and reduced inflammatory cytokines through NRF2 induction. Interestingly, the Nrf2 inhibitor ML385 suppressed the expression of NAD(P)H quinone oxidoreductase 1(NQO1) and enhanced the expression of KEAP1 in HSV-1 infected cells. This led to the reversal of VP16 expression inhibition, a protein factor associated with HSV-1 infection, thereby promoting HSV-1 replication. Conclusion: These findings suggest for the first time that ginsenoside Rg5 may serve as an antiviral against HSV-1 infection and could be a novel therapeutic agent for HSV-1-induced neuroinflammation.

• Molecules and Cells
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• 제26권1호
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• pp.67-73
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• 2008

Herpetic stromal keratitis (HSK) is an inflammatory disorder induced by HSV-1 infection and characterized by T cell-dependent destruction of corneal tissues. It is not known what triggers $CD4^+$ T cell migration into the stroma of HSV-1-infected corneas. The keratocyte is a fibroblast-like cell that can function as an antigen-presenting cell in the mouse cornea by expressing MHC class II and costimulatory molecules after HSV-1 infection. We hypothesized that chemokines produced by stromal keratocytes are involved in $CD4^+$ T cell infiltration into the cornea. We found that keratocytes produce several cytokines and chemokines, including MCP-1, RANTES, and T cell activation (TCA)-3. HSV-1 infection increased the production of MCP-1 and RANTES by keratocytes, and these acted as chemoattractants for HSV-1-primed $CD4^+$ T cells expressing CCR2 and CCR5. ExpreHerpetic stromal keratitis (HSK) is an inflammatory disorder induced by HSV-1 infection and characterized by T cell-dependent destruction of corneal tissues. It is not known what triggers $CD4^+$ T cell migration into the stroma of HSV-1-infected corneas. The keratocyte is a fibroblast-like cell that can function as an antigen-presenting cell in the mouse cornea by expressing MHC class II and costimulatory molecules after HSV-1 infection. We hypothesized that chemokines produced by stromal keratocytes are involved in $CD4^+$ T cell infiltration into the cornea. We found that keratocytes produce several cytokines and chemokines, including MCP-1, RANTES, and T cell activation (TCA)-3. HSV-1 infection increased the production of MCP-1 and RANTES by keratocytes, and these acted as chemoattractants for HSV-1-primed $CD4^+$ T cells expressing CCR2 and CCR5. Expression of MCP-1 in the corneal stroma was confirmed in vivo. Finally, when HSV-1-primed $CD4^+$ T cells were adoptively transferred into wild type and MCP-1-deficient mice that had been sublethally irradiated to minimize chemokine production from immune cells, infiltration of $CD4^+$ T cells was markedly reduced in the MCP-1-deficient mice, suggesting that it is the MCP-1 from HSV-1-infected keratocytes that attracts $CD4^+$ T cells into the cornea.

• Nuclear Medicine and Molecular Imaging
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• 제43권5호
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• pp.468-477
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• 2009

목적: Hydrodynamic-based procedure는 손쉽고 간편한 비바이러스성 유전자 전달 방법으로 특히 간특이적으로 발현하는 특징을 가진다. 단순 헤르페스 바이러스 제 1 형 티미딘 키나제(herpes simplex virus type 1 thymidine kinase, HSV1-tk)와 다양한 기질을 이용한 비침습적 HSV1-tk 유전자 영상시스템이 널리 연구되어왔다. 본 연구에서는 HSV1-tk 유전자를 hydrodynamic-based procedure를 이용하여 전달한 후, HSV1-tk의 보고 기질로 알려진 5-(2-iodovinyl)-2'-deoxyuridine (IVDU)을 이용하여 간 특이적인 HSV1-tk 유전자 발현 영상을 획득하고자 하였다. 대상 및 방법: HSV1-tk 유전자와 녹색형광유전자를 가진 각 플라스미드 벡터를 마우스에 hyodynaminc injection을 통해 전달하고, 24 시간 뒤 유전자의 발현을 확인하기 위해 RT-PCR, 생체형광영상, 핵의학영상, 전신자가방사영상 그리고 생체분포를 시행하였다. 결과: 각 플라스미드 벡터를 전달한 간으로부터 추출한 전체 RNA를 이용하여 RT-PCR을 수행한 결과, 각각 HSV1-tk유전자와 녹색형광단백 유전자의 특이적인 밴드를 관찰할 수 있었다. 생체 분포 결과, pHSV1-tk 벡터를 전달한 마우스의 간에서 특이적인 [$^{123}I$]IVDU의 섭취를 보였다. 생체형광영상에서는pEGFP-N1 벡터를 전달한 마우스의 간에서는 유의한 형광신호를 나타내었다. 전신자가방사영상과 감마카메라 영상에서 pHSV1-tk 벡터를 전달한 마우스의 간에서 방사표지 IVDU가 국소적으로 집적되는 것을 확인하였다. 결론: 본 연구에서 hydrodynamic-based procedure는 간특이적으로 플라스미드 DNA를 전달하는데 효과적이며 전달된 유전자의 발현을 분자영상학적인 방법으로 확인하였다. 따라서 Hydrodynamic injection을 통해 HSV1-tk유전자와 목적 유전자의 공동발현은 방사표지 IVDU에 의해 목적 유전자의 발현을 정량평가하는데 유용할 것으로 기대된다.

• Biomolecules & Therapeutics
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• 제7권2호
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• pp.158-163
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• 1999

In order to find less toxic antiherpetic agents, antiviral activities of quercitrin against two strains of pathogenic viruses such as herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) were determined in Vero cells using plaque reduction assay in vitro. Quercitrin showed a concentration-dependent decrease in plaque formation of HSV-1 and HSV-2. It also exhibited more potent antiherpetic activity on HSV-1 with 50% effective concentration (EC$_{50}$) of 20.4 $\mu$g/ml than on HSV-2 with EC$_{50}$ of 30.4 $\mu$g/ml. The combined antiherpetic effects of quercitrin with nucleoside antiherpetic agents, acyclovir and vidarabine, were examined on the multiplication of these two strains of herpesviruses in Vero cells by the combination assay. The results of combination assay were evaluated by the combination index (CI) that was calculated by the multiple drug effect analysis. The combinations of quercitrin with acyclovir and vidarabine on HSV-1 showed more potent synergism with CI values of 0.27-0.81 for 50%, 70%, 90% effective levels than those on HSV-2 with CI values of 1.03~2.20..20.

• 약학회지
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• 제42권6호
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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.

• BMB Reports
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• 제36권4호
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• pp.379-386
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• 2003

A mutant herpes simplex virus 1, mtHSV, was constructed by inserting the E. coli beta-galactosidase gene into the loci of icp34.5, the apoptosis-inhibiting gene of HSV. The mtHSV replicated in and lysed U251 (human glioma cells), EJ (human bladder cells), and S-180 (mice sarcoma cells), but not Wish (human amnion cells) cells. With its intact tk (thymidine kinase) gene, mtHSV exhibited susceptibility to acyclovir (ACV), which provided an approach to control viral replication. An in vivo test with mtHSV was conducted in immune-competent mice bearing sarcoma S-180 tumors, which were treated with a single intratumoral injection of mtHSV or PBS. Tumor dimensions then were measured at serial time points, and the tumor volumes were calculated. Sarcoma growth was significantly inhibited with prolonged time and reduced tumor volume. There was microscopic evidence of necrosis of tumors in treated mice, whereas no damage was found in other organs. Immunohistochemical staining revealed that virus replication was exclusively confined to the treated tumor cells. HSV-1 DNA was detected in tumors, but not in the other organs by a polymerase chain reaction analysis. From these experiments, we concluded that mtHSV should be a safe and promising oncolytic agent for cancer treatment.