• Journal of Life Science
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• v.17 no.10
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• pp.1361-1367
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• 2007

I investigated the effect of KT5720, an inhibitor of protein kinase A, on the vesicular stomatitis virus (VSV) infection in BHK-21cell cultures. The virus inducted cytopathic effect (CPE) was almost completely suppressed by KT5720 at 5uM. The inhibitor, however, did not affect replication of the virus nor the synthesis of viral macromolecules. KT5720, did not block the cytoskeletal disruption, while the cell rounding was suppressed. And, the KT5720-sensitive function may be involved in developing the VSV-induced CPE, but not essential for the virus replications.

• IMMUNE NETWORK
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• v.1 no.2
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• pp.109-115
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• 2001

Background: The role of the interferon consensus sequence binding protein (ICSBP), a member of interferon regulatory factor family, in protecting against a vesicular stomatitis virus (VSV) infection has not been firmly elucidated. Thus, it was investigated utilizing the human promyelocytic leukemia HL-60 cells which do not express ICSBP. Methods: HL-60 cells were stably transfected with plasmid containing cDNA for either ICSBP or DNA binding domain (DBD) and tested for their VSV-susceptibilities. The susceptibility of each transfectant group to a VSV infection was determined by a plaque assay at 1 h, 24 h, and 48 h post-infection in the presence (500 IU/ml) or absence of interferon ${\alpha}$ ($IFN{\alpha}$). Results: In the absence of $IFN{\alpha}$, the three groups showed similar sensitivities to a VSV infection. However, when pre-treated with IFN, the viral titers in both the ICSBP and control clones steadily decreased over 48 h of incubation, indicating the existence of $IFN{\alpha}$-mediated protection against VSV infection. The $IFN{\alpha}$-treated ICSBP clones appeared to be more resistant to infection compared with the control clones, although the difference was not great. On the contrary, the viral titers in the $IFN{\alpha}$-treated DBD clones increased at 24 h then decreased by 48 h. Conclusion: The expression of truncated ICSBP (DBD) does not appear to underlie the impaired protection against a VSV infection in the DBD clones, since even the control clones lacking ICSBP were protected from a VSV infection. This suggests that ICSBP does not play a critical role in the $IFN{\alpha}$- mediated anti-VSV response of HL-60 cells, although it appears to confer some resistance to a VSV infection.

• KSBB Journal
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• v.22 no.5
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• pp.282-287
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• 2007

The antiviral mechanism of KT5720 is known to inhibit the cAMP-dependent protein kinase (PKA), on the VSV infection in BHK-21 cell cultures. The virus inducted CPE (cell rounding) was almost completely suppressed by KT5720 at 5 uM. The inhibitor, however, did not affect the replication of the virus and the synthesis of viral macromolecules. Immunological studies showed the viral matrix (M) protein displayed intimate association with the cytoskeletal components and probably the cell rounding. KT5720, did not block the cytoskeletal disruption, while the cell rounding was suppressed. These observations suggest that the interaction between the viral M protein and the cytoskeletal components may not be enough to cause the morphological change of the cell. And, the KT5720-sensitive function may be involved in developing the VSV-induced CPE, but not essential for the virus replications.

• Journal of fish pathology
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• v.30 no.2
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• pp.71-78
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• 2017

The glycoprotein of novirhabdoviruses is known to play a critical role in the determination of host specificity. Viral hemorrhagic septicemia viruses (VHSVs) in different genotypes have different glycoprotein sequences and show different preferences for specific cell lines. In this study, to know whether the glycoprotein is solely responsible for the host cell preference of VHSV, a recombinant VHSV expressing vesicular stomatitis virus (VSV) glycoprotein instead of VHSV IVa glycoprotein (rVHSV-VSV-G) was generated by reverse genetics and inoculated into several fish cell lines, then, cytopathic effect (CPE) and viral growth caused by rVHSV-VSV-G infection were compared with those caused by rVHSV-wild that was previously generated and has the same genomic sequence with wild-type VHSV except a few nucleotides. The plaque numbers of rVHSV-VSV-G were significantly higher in EPC, BF-2 and GF cells than those of rVHSV-wild. However, in HINAE cells (originated from olive flounder), rVHSV-VSV-G titer was significantly lower than rVHSV-wild titer, and both recombinant VHSVs were not grown well in CHSE-214 cells. Although statistical significances were detected in the titers between rVHSV-wild and rVHSV-VSV-G in several cell lines, the cell line-preference order of rVHSV-VSV-G was not different from that of rVHSV-wild. These results suggest that the replacement of VHSV glycoprotein may not completely change host cell preference, and other regions of VHSV might also involve in the determination of host cell preference.

• Proceedings of the Microbiological Society of Korea Conference
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• 2000.10a
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• pp.66-77
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• 2000

For a variety of viruses, the primary virus infection has been shown to prevent superinfection with a homologous secondary virus; however, the mechanism of exclusion has not been clearly understood. In this work, we demonstrated that BVDV -infected MDBK cells were protected from superinfection with a homologous superinfecting BVDV, one of the positive-sense RNA pestiviruses, but not with an unrelated rhabdovirus, such as vesicular stomatitis virus. Once superinfection exclusion was established by a primary infection with BVDV, the transfected infectious BVD viral RNA genome was shown to be competent for viral translation, but not viral replication. In addition, our results also demonstrated that upon superinfection, the. viral RNA genome of viral particles was not transferred into the cytoplasm of BVDV -infected cells. Using newly developed system involving rapid generation of the MDBK cells expressing BVD viral proteins, we subsequently found that expression of the viral structural proteins was dispensable for the block occurring at the level of viral RNA replication, but required for the exclusion at the level of viral entry step. Altogether, these findings provide evidence that the superinfection exclusion of BVDV occurs not only at the level of viral replication in which the viral replicase are involved, but also at the level of viral entry with which the viral structural proteins are associated, and that a cellular factor(s) play an essential role in this process.

• Journal of Embryo Transfer
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• v.1 no.1
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• pp.16-27
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• 1986

Based on the current importing and exporing regulations for disease control of embryo transfer, some important microorganisms and their control possibilities are reviewed. The results reviewed were sumrnarized as follows: 1. Regulations regarding to the import of embryos vary between importing and exporting countries, but exporting countries examine the donor and embryos for the heaith certification by the requirements of importing countries. 2. Organisms that infect the gametes are 5 kinds of viruses and the diseases caused by them could not be controlled or eradicated using embryo transfer. 3. Organisms that do not infect the gametes are 4 kinds of viruses and the causal organisms are potential candidates for control or eradication by embryo transfer. 4. Organisms that penetrate the zona pellucida and infect the embryo are 6 kinds of viruses including bovine viral diarrhea virus. 5. Organisms that cannot penetrate the zona pellucida or do not infect the embryo are 15 kinds of viruses and the removal from their contaminations are recommended by proper washing procedure and antisera treatment. Bovine and porcine parvovirus, porcine pseudorabies virus and vesicular stomatitis virus are included in these organisms. 6. Bovine embryos that artificially exposed to various pathogenic organisms such as bovine herpes virus, IBR virus, bluetongue virus, bovine viral diarrhea virus and Brucella abortus in vitro are discussed about their infection by several treatments.

• BMB Reports
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• v.34 no.6
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• pp.566-572
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• 2001

Chimeric CD4 proteins were assembled. They contained the entire CD4 ectodomain that is linked to different membrane anchors. Membrane anchors consisted of either glucosyl phosphatidyl inositol (gpi), the transmembrane and cytoplasmic regions of HIV-1 Env protein, or the vesicular stomatitis virus G glycoprotein, respectively. The HIV-1 co-receptor CXCR4 and CD4 were independently inserted into viral envelopes. We compared the insertion of six different CD4/CXCR4 constructs into HIV-1 envelopes, as well as their functionality in targeting and specific infection of cells that constitutively express the HIV-1 Env protein. All of the six different HIV-1 (CD4/CXCR4) pseudotypes were able to transduce Env (+) cells at similar efficiency. In addition, stable transduction of the Env (+) recipient cells demonstrated that all chimeric proteins were functional as receptors for Env when inserted into HIV-1 envelopes. In fact, these results demonstrate for the first time a stable transduction by a targeted HIV-1 pseudotype virus.

• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.49-49
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• 2003

본 연구에서는 vesicular stomatitis virus G glycoprotein (VSV-G)를 envelope로 가지는 pantropic retrovirus vector system을 이용하여 재조합 human FSH 유전자가 전이된 형질전환 닭을 생산하고자 하였다. Human FSH $\alpha$ 및 $\beta$ 유전자와 CTP linker는 human pituitary gland cDNA library에서 RT-PCR 방법을 이용하여 cloning하였으며, 각각의 fragment는 FSH$\beta$-CTP-FSH$\alpha$ 순서의 단일사슬로 연결하였다. 연결된 FSH$\beta$-CTP-FSH$\alpha$는 retroviral vector 내의 $\beta$-actin promoter의 조절 하에 도입한 후, PT67 packaging cell line에 transfection하여 virus를 생산하였으며 생산된 virus는 pantropic한 virus producing cell인 GP293에 infection하여 FSH 유전자가 도입된 virus를 생산하였다. FSH 유전자의 발현을 in vitro에서 확인하기 위하여 CHO (chinese hamster ovary) 세포에 virus를 감염시킨 후, 세포의 배양액을 취하여 electrochemilumine-scence immunoassay 방법으로 정량하였다. In vitro에서 전이 후 발현이 확인된 FSH 외래유전자의 retroviral vector virus를 초원심분리로 고농축하여 stageX의 계란의 배반엽 층에 주입하였으며, 그 결과 18%의 부화율과 91%의 부화한 닭의 유전자 전이율을 확인할 수 있었다. 전이된 유전자의 확인은 FSH$\beta$와 Neo 유전자에 대한 primer를 이용한 RT-PCR의 방법을 이용하였다. In vitro에서와는 달리 in vivo에서는 FSH 유전자의 전이는 확인되었으나 발현을 확인하지는 못하였는데, 이는 적은 수의 실험군이 형질전환율에 비해 상대적이지 못하였거나, 외래 유전자인 FSH의 발현에 의한 생리적인 부작용이 유발되어 해당개체가 부화되지 못한 것으로 추정된다. 본 문제점을 해결하기 위하여 실험군의 수를 늘리고 외래 유전자에 대한 controllable expression system이 보완될 필요성이 요구되며, 이러한 점이 해결된다면 높은 유전자 전이율에 기인하여 retrovirus를 이용한 형질전환 방법은 형질전환 가금의 생산에 있어서 매우 효율적이고 주목할 만한 방법으로 사료된다.

• Journal of Veterinary Clinics
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• v.27 no.3
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• pp.268-272
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• 2010

South Korea is susceptible to foreign diseases due to its high rate of livestock importation. The purpose of this study was to investigate the infectious conditions of contagious disease of horses imported into South Korea from other countries. The horses were tested for contagious equine metritis (CEM), equine infectious anemia (EIA), equine piroplasmosis (EP), equine viral arteritis (EVA), vesicular stomatitis (VS), dourine, and glanders. The prevalence of these infectious diseases in 6,650 horses imported from 24 countries between 2003 and 2008 was reviewed by the National Veterinary Research and Quarantine Service. Seropositive results were found for EIA, EP, EVA, dourine and glanders: 3/6,189 serum samples tested were EIA-positive, 37/6,005 samples tested by complement fixation (CF) were EP-positive, 28/6,043 samples tested by virus neutralization (VN) were EVA-positive, 4/2,071 serum samples tested by CF were positive for dourine, and 4/1,950 samples tested by CF were positive for glanders. No contagious equine metritis or vesicular stomatitis was detected. In total, 76/6,650 imported horses tested positive for an infectious disease. Notably, 4/6 sera (66.6%), all taken from horses imported from Tanzania, were positive for glanders. This is the first report of glanders infection in horses from Tanzania since 1996.

• IMMUNE NETWORK
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• v.11 no.3
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• pp.155-162
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• 2011

Background: Toll-like receptor 3 (TLR3) recognizes double-stranded RNA (dsRNA) and induces inflammation. In this study we attempted to ascertain if there are endogenous host molecules controlling the production of cytokines and chemokines. Two candidates, ribosomal protein L19 and L22, were analyzed to determine if they influence cytokine production followed by TLR3 activation. In this study we report that L19 acts upon production of IP-10 or IL-8 differently in glioblastoma cells. Methods: L19 or L22 was transfected into HEK293-TLR3, A549 or A172 cells. After treatment with several inhibitors of NF-${\kappa}B$, PI3K, p38 or ERK, production of IL-8 or IP-10 was measured by ELISA. siRNA was introduced to suppress expression of L19. After Vesicular stomatitis virus infection, viral multiplication was measured by western blot. Results: L19 increased ERK activation to produce IL-8. In A172 cells, in which TLR3 is expressed at endosomes, L19 inhibited interferon regulatory factor 3 (IRF3) activation and IP-10 production to facilitate viral multiplication, whereas L19 inhibited viral multiplication in A549 cells bearing TLR3 on their cell membrane. Conclusion: Our results suggest that L19 regulates TLR3 signaling, which is cell type specific and may be involved in pathogenesis of autoimmune diseases and chronic inflammatory diseases.