• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.137.1-137
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• 2003

A pepper strain of Cucumber mosaic virus (Pf-CMV) induces a mild chlorotic spot symptom in zucchini squash at 9 days post-inoculation (dpi), wile Fny strain of CMV causes severe mosaic and stunting symptom at 4 dpi in this host. Pseudorecombinants were constructed between the two strains, and assessments of symptom severity were indicated that both RNA2 and RNA3 were responsible for both mildness and the slow appearance of symptom elicited by Pf-CMV in zucchini squash. With various RNA2 and RNA3 chimeras between two strains of CMV, the genetic symptom determinants of phenotype of Pf-CMV were mapped to Tyr residue at positions amino acid 267 in 2a protein and at positions amino acid 168 in 3a movement protein (MP). Chimeras changed the sequences (both changed Tyr to lie) in the codons of both amino acid 168 of 3a MP and amino acid 267 of 2a protein were resulted in the high RNA accumulation, severity of symptom, and the rapid systemic spread, suggesting that 2a replicase as well as MP is involved in virus movement. The RNA accumulation pattern of all pseudorecombinants and chimeras are identical in protoplast of zucchini squash, indicating the virus movement is responsible for the phenotypes of two CMV strains rather than virus replication.

• Molecules and Cells
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• v.43 no.5
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• pp.469-478
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• 2020

Hepatitis C virus (HCV) propagation is highly dependent on cellular proteins. To identify the host factors involved in HCV propagation, we previously performed protein microarray assays and identified the LIM and SH3 domain protein 1 (LASP-1) as an HCV NS5A-interacting partner. LASP-1 plays an important role in the regulation of cell proliferation, migration, and protein-protein interactions. Alteration of LASP-1 expression has been implicated in hepatocellular carcinoma. However, the functional involvement of LASP-1 in HCV propagation and HCV-induced pathogenesis has not been elucidated. Here, we first verified the protein interaction of NS5A and LASP-1 by both in vitro pulldown and coimmunoprecipitation assays. We further showed that NS5A and LASP-1 were colocalized in the cytoplasm of HCV infected cells. NS5A interacted with LASP-1 through the proline motif in domain I of NS5A and the tryptophan residue in the SH3 domain of LASP-1. Knockdown of LASP1 increased HCV replication in both HCV-infected cells and HCV subgenomic replicon cells. LASP-1 negatively regulated viral propagation and thereby overexpression of LASP-1 decreased HCV replication. Moreover, HCV propagation was decreased by wild-type LASP-1 but not by an NS5A binding-defective mutant of LASP-1. We further demonstrated that LASP-1 was involved in the replication stage of the HCV life cycle. Importantly, LASP-1 expression levels were increased in persistently infected cells with HCV. These data suggest that HCV modulates LASP-1 via NS5A in order to regulate virion levels and maintain a persistent infection.

• Korean journal of applied entomology
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• v.43 no.3 s.136
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• pp.225-231
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• 2004

An endoparasitoid wasp, Cotesia plutellae, has been used for a biological control agent against the diamondback moth, Plutellae xylostella. It has a symbiotic polydnavirus in their reproductive tract, which is required for its successful parasitization. Here, we measured a specific replication time of the polydnavirus during female development of C. plutellae. We, also, analyzed the reproductive potentials of female C. plutellae under mating or different host conditions. At $25^{\circ}C$, pupal C. plutellae began to develop adult tissues such as compound eyes and wings since day 2. At day 5, all adult tissues including antennae were developed and were ready to emerge. With polyclonal antibody raised against C. plutellae polydnavirus, an immunobloting could confirm virus replication at day 4 during pupal stage. Virus particles could be visualized by transmission electron microscope in the oviduct lumen of day 5 pupae. After adult eclosion, venom gland and ovarian calyx increased in size, though ovarioles did not. Mated females layed large number of eggs (over $60\%$) at first 4 days during their mean longevity of ca. 8 days at $25^{\circ}C$. Unmated females showed less active ovipositional behavior, where all the eggs developed into males. C. Plutellae parasitized both P. xylostella and fall webworm, Hyphantria cunea. However, C. Plutellae developed faster and showed higher successful paarasitization in P. xylostella than in H. cunea.

• Biomolecules & Therapeutics
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• v.32 no.4
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• pp.481-491
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• 2024

Paxlovid is the first approved oral treatment for coronavirus disease 2019 and includes nirmatrelvir, a protease inhibitor targeting the main protease (M^pro) of SARS-CoV-2, as one of the key components. While some specific mutations emerged in M^pro were revealed to significantly reduce viral susceptibility to nirmatrelvir in vitro, there is no report regarding resistance to nirmatrelvir in patients and animal models for SARS-CoV-2 infection yet. We recently developed xenograft tumors derived from Calu-3 cells in immunodeficient mice and demonstrated extended replication of SARS-CoV-2 in the tumors. In this study, we investigated the effect of nirmatrelvir administration on SARS-CoV-2 replication. Treatment with nirmatrelvir after virus infection significantly reduced the replication of the parental SARS-CoV-2 and SARS-CoV-2 Omicron at 5 days post-infection (dpi). However, the virus titers were completely recovered at the time points of 15 and 30 dpi. The virus genomes in the tumors at 30 dpi were analyzed to investigate whether nirmatrelvir-resistant mutant viruses had emerged during the extended replication of SARS-CoV-2. Various mutations in several genes including ORF1ab, ORF3a, ORF7a, ORF7b, ORF8, and N occurred in the SARS-CoV-2 genome; however, no mutations were induced in the M^pro sequence by a single round of nirmatrelvir treatment, and none were observed even after two rounds of treatment. The parental SARS-CoV-2 and its sublineage isolates showed similar IC₅₀ values of nirmatrelvir in Vero E6 cells. Therefore, it is probable that inducing viral resistance to nirmatrelvir in vivo is challenging differently from in vitro passage.

• Biomolecules & Therapeutics
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• v.3 no.3
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• pp.210-216
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• 1995

Partially purified water-soluble extract mixture from Ricini and Coptidis (named as RIC) showed to be a potent inhibitor of human immunodeficiency virus-1 (HIV-1) replication. RIC was evaluated for in vitro anti-HIV activity using SupTl and H9 cells infected by a recombinant virus (pSVCAT) containing chloramphenicol acetyltransferase (CAT) gene substituted for nef gene in the HIV-1 genome. RIC inhibited syncytiaformation of SupTl cells with a half maximal effective concentration, $IC_{50}$/, of 2.5 $\mu\textrm{g}$/mι and showed marked inhibition of CAT activity in the infected H9 cells and also suppressed reverse transcriptase (RT) activity in the supernatant of the infected H9 culture. However, RIC did not inhibit the activity of reverse transcriptase directly when it was mixed with the enzyme or with viral particles. Berberine, one of components of RIC, also showed similar anti-HIV activity as RIC did. The data suggest that there are active ingredients which mediate anti-HIV activity in RIC.

• YAKHAK HOEJI
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• v.57 no.1
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• pp.43-54
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• 2013

Viral hepatitis is the inflammation of liver cells caused by viruses, and still one of the major health-care problems worldwide. A number of viruses to cause hepatitis are type A, B, C, D, E or G. Among these viruses leading to hepatitis, B and C are more troublesome being more prone to chronic illness which can cause the potentially fatal conditions of hepatocellular carcinoma (HCC) and/or liver failure. If immediate treatment is not initiated, liver transplant is the only option left. Over the past few decades there has been remarkable progress in diagnose and monitor all hepatitis virus infections for treatment and prevention. Nonetheless, important challenges remain to develop more effective and safe vaccines for prevention as well as antiviral agents to reduce viremia/viral load by inhibiting viral replication. The development and evaluation of antiviral agents through carefully designed clinical trials over the last 25 years has heralded a new dawn in the treatment of patients chronically infected with the hepatitis B and C viruses, but not so for the D virus. The introduction of Direct Acting Antivirals (DDAs) for the treatment of HBV carriers has permitted the long term use of these compounds for the continuous suppression of viral replication. This review aims to summarize the current status and development approaches of antiviral drugs for the treatment of viral hepatitis and future perspectives.

• Journal of Veterinary Science
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• v.21 no.5
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• pp.72.1-72.10
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• 2020

Background: Fenbendazole, a dewormer drug, is used widely in the clinical treatment of parasite infections in animals. Recent studies have shown that fenbendazole has substantial effects on tumor growth, immune responses, and inflammatory responses, suggesting that fenbendazole is a pluripotent drug. Nevertheless, the antiviral effects have not been reported. Fenbendazole can disrupt microtubules, which are essential for multiple viruses infections, suggesting that fenbendazole might have antiviral effects. Objectives: This study examined whether fenbendazole could inhibit bovine herpesvirus 1 (BoHV-1) productive infection in cell cultures. Methods: The effects of fenbendazole on viral production, transcription of the immediate early (IE) genes, viron-associated protein expression, and the cellular signaling PLC-γ1/Akt pathway were assessed using distinct methods. Results: Fenbendazole could inhibit BoHV-1 productive infections significantly in MDBK cells in a dose-dependent manner. A time-of-addition assay indicated that fenbendazole affected both the early and late stages in the virus replication cycles. The transcription of IE genes, including BoHV-1 infected cell protein 0 (bICP0), bICP4, and bICP22, as well as the synthesis of viron-associated proteins, were disrupted differentially by the fenbendazole treatment. The treatment did not affect the cellular signaling pathway of PLC-γ1/Akt, a known cascade playing important roles in virus infection. Conclusions: Overall, fenbendazole has antiviral effects on BoHV-1 replication.

• Journal of Microbiology and Biotechnology
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• v.18 no.7
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• pp.1326-1334
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• 2008

The prime-boost vaccination with DNA vaccine and recombinant viral vector has emerged as an effective prophylactic strategy to control infectious diseases. Here, we compared the protective immunities induced by multiple alternating immunizations with DNA vaccine (pCIgB) and replication-incompetent adenovirus (Ad-gB) expressing glycoprotein gB of pseudorabies virus (PrV). The platform of pCIgB-prime and Ad-gB-boost induced the most effective immune responses and provided protection against virulent PrV infection. However, priming with pCIgB prior to vaccinating animals by the DNA vaccine-prime and Ad-boost protocol provided neither effective immune responses nor protection against PrV. Similarly, boosting with Ad-gB following immunization with DNA vaccine-prime and Ad-boost showed no significant responses. Moreover, whereas the administration of Ad-gB for primary immunization induced Th2-type-biased immunity, priming with pCIgB induced Th1-type-biased immunity, as judged by the production of PrV-specific IgG isotypes and cytokine IFN-$\gamma$. These results indicate that the order and injection frequency of vaccine vehicles used for heterologous prime-boost vaccination affect the magnitude and nature of the immunity. Therefore, our demonstration implies that the prime-boost protocol should be carefully considered and selected to induce the desired immune responses.

• BMB Reports
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• v.37 no.3
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• pp.383-385
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• 2004

RNA interference (RNAi) is a process of sequence-specific gene silencing, which is initiated by double-stranded RNA (dsRNA). RNAi may also serve as an antiviral system in vertebrates. This study describes the inhibition of herpesvirus-6B (HHV-6B) replication by short interference RNAs (siRNAs) that are targeted to the U38 sequence that encodes DNA polymerase. When virus-infected SupT1 cells were treated by siRNA, these cells blocked the cytopathic effect (CPE) and detected the HHV-6B antibody-negative in indirect immunofluorescence assays (IFA). Our result suggests that RNAi can efficiently block Herpesvirus-6B replication.

• Journal of Microbiology and Biotechnology
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• v.24 no.12
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• pp.1744-1754
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• 2014

The hepatitis C virus (HCV) RNA genome is replicated by an RNA replicase complex (RC) consisting of cellular proteins and viral nonstructural (NS) proteins, including NS5B, an RNA-dependent RNA polymerase (RdRp) and key enzyme for viral RNA genome replication. The HCV RC is known to be associated with an intracellular membrane structure, but the cellular components of the RC and their roles in the formation of the HCV RC have not been well characterized. In this study, we took a proteomic approach to identify stomatin, a member of the integral proteins of lipid rafts, as a cellular protein interacting with HCV NS5B. Co-immunoprecipitation and co-localization studies confirmed the interaction between stomatin and NS5B. We demonstrated that the subcellular fraction containing viral NS proteins and stomatin displays RdRp activity. Membrane flotation assays with the HCV genome replication-competent subcellular fraction revealed that the HCV RdRp and stomatin are associated with the lipid raft-like domain of membranous structures. Stomatin silencing by RNA interference led to the release of NS5B from the detergent-resistant membrane, thereby inhibiting HCV replication in both HCV subgenomic replicon-harboring cells and HCV-infected cells. Our results identify stomatin as a cellular protein that plays a role in the formation of an enzymatically active HCV RC on a detergent-resistant membrane structure.