• Title/Summary/Keyword: virus replication

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The Interaction between HCV-Infected huh7.5 Cells and HCV-Specific T Cells (C형 간염 바이러스 감염 간암 세포주와 T 림프구의 상호작용에 대한 연구)

  • Kang, Hyojeung;Cho, Hyosun
    • Korean Journal of Microbiology
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    • v.50 no.2
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    • pp.169-172
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    • 2014
  • Recently, Hepatitis C virus (HCV) replication system has been established using human hepatoma cells (huh cell) and a variety of HCV clones. In this study, we established an infectious HCV replication system using huh7.5 cells and J6/JFH1 clone (genotype 2a). In addition, we investigated the antigen presentation capability of HCV-infected huh7.5 cells to HCV-specific T cells. Interestingly, HCV-infected huh7.5 cells were not capable of activating HCV-specific T cells. However, huh7.5 cells stimulated by exogenous HCV peptide were able to activate HCV-specific T cells, which was shown to produce TNF-${\alpha}$ and IFN-${\gamma}$. We further examined if HCV infection has an inhibitory effect on the expression of MHC class I molecule of huh7.5 cells. We found that HCV infection did not change the expression level of MHC class I molecule on huh7.5 cells.

Induction of antiviral mechanisms by interferon-related genes in rock bream (Oplegnathus fasciatus) infected with rock bream iridovirus (RBIV)

  • Myung-Hwa Jung
    • Journal of fish pathology
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    • v.36 no.2
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    • pp.213-228
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    • 2023
  • We evaluated the transcriptional response of interferon (IFN)-related genes in rock bream iridovirus (RBIV)-infected rock bream under high-, low-, or no-mortality conditions induced by different stocking water temperatures. Under the high susceptibility condition (group A, water temperature 26℃, 100% mortality), only the Mx gene was expressed early, with prolonged expression, and with heavy viral loads of approximately 106~107 major capsid protein gene copies/μL from 4 to 10 days post infection (dpi). However, IRF1, IRF3, IRF8, STAT1, ISG15, PKR, Viperin, GVIN1, IFI44, and ISG56 were activated at later time points (8 dpi) and then quickly decreased (10 dpi). For the low susceptibility condition, the water temperature was set at 23℃ for 7 days (group B) and then reduced to 17℃. Group B exhibited a 28% mortality rate, in which persistent and effective antiviral responses were observed for long periods of time. In particular, at 20 and 22 dpi, when virus replication was peaked at approximately 107/μL, the expressions of most of the IFN-related genes (IRF1, IRF3, IRF8, Mx, STAT1, ISG15, PKR, Viperin, GVIN1, IFI44, and ISG56) were significantly higher in group B than in the control group. Moreover, prolonged and higher levels of IRF3 (at least 30 dpi), IRF8 (at least 30 dpi), ISG15 (at least 30 dpi), PKR (at least 28 dpi), Viperin (at least 30 dpi), and IFI44 (at least 30 dpi) were also observed in the recovery stage of infection. Under the no-susceptibility condition at 17℃ (0% mortality), significantly elevated levels of IRF3, Mx, ISG15, and PKR were observed mostly until 20 dpi. The findings indicate that RBIV infection can induce an efficient IFN-mediated antiviral immune response in low- and no-susceptibility conditions. The findings could be valuable for effective control of viral pathogens in fish.

Generation of Transgenic Chickens that Produce Bioactive Human Thrombopoietin (재조합 hTPO를 생산하는 형질전환 닭의 개발)

  • Kwon, Mo-Sun;Koo, Bon-Chul;Roh, Ji-Yeol;Lee, Hyun-A;Kim, Te-Oan
    • Reproductive and Developmental Biology
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    • v.32 no.3
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    • pp.159-166
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    • 2008
  • We report here the generation of transgenic chickens that produce human Thrombopoietin (hTPO) using replication-defective Moloney murine leukemia virus (MoMLV)-based vectors packaged with vesicular stomatitis virus G glycoprotein (VSV-G). For the retrovirus vectors, we used hCMV (human Cytomegalovirus) internal promoter to drive the hTPO gene. After confirming the expression of the hTPO gene in various target cells, the concentrated solution of recombinant retrovirus was injected beneath the blastoderm of non-incubated chicken embryos (stage X). The biological activity of the recombinant hTPO in target cell was significantly higher than its commercially available counterpart. Out of 132 injected eggs, 11 chicks hatched after 21 days of incubation and 4 hatched chicks were found to express vector-encoded hTPO gene. However, 3 out of the 4 transgenics died within one month of hatching. The major significance of this study is that it is one of the very few successful reports on the production of transgenic chickens as bioreactors aiming mass production of commercially valuable and biological active human cytokine proteins.

Studies on the rabbit viral hepatitis I. Electron microscopic observation of the acute hepatic lesions in experimentally infected rabbit (토끼의 바이러스성(性) 간염(肝炎)에 관(關)한 연구(硏究) I. 실험적(實驗的) 감염토(感染兎)의 급성간염조직(急性肝炎組織)의 전자현미경적(電子顯微鏡的) 관찰(觀察))

  • Lee, Cha-soo;Park, Cheong-kyu
    • Korean Journal of Veterinary Research
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    • v.29 no.4
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    • pp.531-540
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    • 1989
  • A new sudden death in rabbits appeared in China and Korea in 1984 and 1985, respectively, and was recognized to be an acute infectious disease caused by a virus. The disease was reported as a "new viral disease," and thereafter, a tentative name of "viral hemorrhagic disease", "hemorrhagic pneumonia" or "viral hemorrhagic pneumonia" has been described in the case reports. But authors had called the viral disease "rabbit viral hepatitis" due to picornavirus infection, because the principal lesion of the disease was an acute hepatitis. The purpose of this report is to describe the electron microscopic findings on the livers in experimentally infected rabbits. All the livers of the affected rabbits were shown to have degenerative changes of a type that is characteristic of acute hepatitis. In the liver cells, there were dilation of rER and mitochondria, vacuole formation of various sizes, and appearances of many virus-like particles in the vicinity of rER, granular bodies and crystalline arrays of viral particles in the cytoplasm with necrotic changes of the nucleus. Clusters of virus-like particles and viral crystals appeared in the cytoplasm of sinusoid endothelial cells and Kupffer's cells with morphological changes of organelles. Also viral crystals were demonstrated in the cytoplasm of macrophages among the liver cells. On the whole, the liver cells had many virus-like particles and a few crystalline arrays of viral particles. Therefore, this implies that the liver cells are the main site of the viral replication in inducing the viremia. It was concluded that the liver was the primary target organ of this viral disease, and the pathological and the ultrastructural evidence suggest that the virus may be belong to genus enterovirus.

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Effects of Fetal Bovine Serum Concentration on the Propagation of Korean Viral Haemorrhagic Septicaemia Virus in an Epithelioma Papulosum Cyprini Cell Line (우리나라에서 분리된 바이러스성출혈성패혈증 바이러스(Viral Haemorrhagic Septicaemia Virus, VHSV)의 증식에 대한 Fetal Bovine Serum (FBS) 농도의 영향)

  • Kim, Jin Hui;Park, Jeong Su;Kwon, Se Ryun;Kim, Shin Hu;Kim, Hyoung Jun
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.51 no.1
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    • pp.42-46
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    • 2018
  • Fetal bovine serum (FBS) is essential for cell culture and is used in the determination of infectivity titer and propagation of viruses. To clarify the effects of FBS on the propagation of viral haemorrhagic septicaemia virus (VHSV), which is a causative agent of mass mortalities of olive flounder Paralichthys olivaceus in Korea, VHSV was inoculated into an EPC (epithelioma papulosum cyprinid) cell line supplemented with MEMs (minimal essential medium) with FBS concentrations of 0%, 2%, 5%, and 10% (MEM0, MEM2, MEM5, and MEM10), respectively, and infectivity titers were compared. Cytopathic effects were observed in all experimental groups at 2 days post virus inoculation (dpi) and all cells were detached from cell culture flasks at 7 dpi. Infectivity titers increased to 3 dpi, persisted to 7 dpi, and decreased when cells were detached. The titer of VHSV in EPC cells in MEM0 was the lowest while those in the other experimental groups showed similar levels. In conclusion, 2% (v/v) of FBS was sufficient to propagate VHSV in EPC cells and the withdrawal of VHSV from cell culture flasks should be performed before cell detachment.

Finding and Characterization of Viral Nonstructural Small Protein in Prospect Hill Virus Infected Cell

  • Nam, Ki-Yean;Chung, Dong-Hoon;Choi, Je-Won;Lee, Yun-Seong;Lee, Pyung-Woo
    • The Journal of Korean Society of Virology
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    • v.29 no.4
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    • pp.221-233
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    • 1999
  • Prospect Hill Virus (PHV) is the well known serotype of hantavirus, a newly established genus in family Bunyaviridae. Extensive studies have upheld the original view of PHV genetics with three genes such as nucleocapsid (N) protein, envelope proteins (G1, G2) and RNA dependent RNA polymerase. In this study, we report the existence of additional gene that is encoded in an overlapping reading frame of the N protein gene within S genome segment of PHV. This gene is expected to encode a nonstructural small (NSs) protein and it seems to be only found in PHV infected cell. The presence and synthesis of NSs protein could be demonstrated in the cell infected with PHV using anti-peptide sera specific to the predicted amino acid sequence deduced from the second open reading frame. Ribosomal synthesis of this protein appears to occur at AUG codon at the 83rd base of S genome segment, downstream of N protein initiation codon. This protein is small in size (10.4 KDa) and highly basic in nature. The expression strategy of NSs protein appears that a signal mRNA is used to translate both N and NSs protein in PHV infected cell. 10 KDa protein in virus infected cell lysates can bind to mimic dsRNA. This fact strongly suggests that NSs protein may be involved in virus replication on late phase of viral life cycle.

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Study on the pathogenesis of the piglets experimentally infected with Korean isolate of Aujeszky′s disease virus I. Histopathologic and electron microscopic observation (Aujeszky's disease virus 국내분리주 접종자돈의 병리발생에 관한 연구 I. 병리학적 및 전자현미경적 관찰)

  • 조우영;조성환;김재훈;박최규;황의경;조부제;정운선
    • Korean Journal of Veterinary Service
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    • v.19 no.1
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    • pp.1-29
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    • 1996
  • This study was conducted to elucidate the pathogenesis of Aujeszky's disease virus(ADV) by histopathologic examination. The first Korean ADV Isolate, which was isolated from piglets with clinical signs of Aujeszky's disease in Yangsan(YS) county, Kyungnam province, was inoculated into 32 days old piglets with a dose of $10^{5.9}$$TCID_{50}/ml$ through intranasal or intramuscular route. These piglets were sacrificed at intervals of every 24hrs for 8 days. The virulence of YS strain was determined by the observation of clinical signs, gross findings, and histopathologic changes in tissues. The virus recovery test was performed from brain, spleen, lung and tonsil in cell culture. The pathogenesis of YS strain was determined by the observation of histopathologlc lesions in CNS and neuronal tracts. The major clinical signs were fever, anorexia, dyspnea, constipation, tremor, ataxia, circling movement, hindleg paralysis and salivation. The clinical signs were more severe in piglets of the group inoculated intranasally than those of the intramuscularly inoculated gorup. Lymphocytopenia was detected on day 5 to day 6 postinoculation (PI). The ADV was recovered from the tissue homogenates of tonsil, lung, spleen and cerebrum in cell culture. The highest virus titer was detected from tonsil between day 6 and day 7 PI. Reddish sublobar consolidation foci were scattered in the apical and cardiac lobes of lung. Although yellowish necrotic foci were detected in tonsil and liver, hemorrhagic lesions were mainly observed in heart, kidney and lymph nodes. Histopathologically, degeneration and necrosis of nerve cells, nonsuppurative meningoe-ncephalitis, nodular gliosis and perivascular cuffings were observed in CNS. Multifocal fibronecrotic foci were observed in lung, liver, lymph nodes and spleen. The major pathologic changes were detected in the midbrain, pons and medulla oblongata. Eosinophilic intranuclear inclusion bodies were mainly observed in epithelia and /or macrophages of tonsil, liver, lung, spleen and submandibular lymph nodes, and neurons of brain, respectively. Observation of viral particles at various stages of replication were possible from the endothelial cells of the alveolar capillaries and tonsillar crypt epithelia by transmission electron microscope.

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Analysis of Syncytium Formation Mechanism induced by Ecotropic Murine Retrovirus (마우스레트로바이러스에 의한 합포체 형성 기작 분석)

  • Bae, Eun-Hye;Park, Sung-Han;Jung, Yong-Tae
    • Korean Journal of Microbiology
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    • v.42 no.3
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    • pp.230-234
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    • 2006
  • To study the mechanism of syncytium formation, novel syncytia-inducing ecotropic murine retrovirus was used. Our previous result showed that amino acid substitutions at the RBD (receptor binding domain) of envelope glycoprotein contribute to syncytium formation. In this study, we have investigated if this fusion phenomenon could occur with retroviral vectors pseudotyped with the novel syncytia-inducing ecotropic murine leukemia virus Env. We have found that these vectors were not able to mediate virus-to-cell fusion in M. dunni murine cell lines. These findings indicate that syncytia-inducing ecotropic murine leukemia virus is capable of generating syncytia during its replication. There was also no correlation between the level of ecotropic murine leukemia virus receptor (mCAT-1) and the fusogenic effect.

Screening of Antiviral Medicinal Plants against Avian Influenza Virus H1N1 for Food Safety

  • Lee, Jang-Hyun;Van, Nguyen Dinh; Ma, Jin-Yeul;Kim, Young-Bong;Kim, Soo-Ki;Paik, Hyun-Dong
    • Food Science of Animal Resources
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    • v.30 no.2
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    • pp.345-350
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    • 2010
  • Various extracts from 30 medicinal plants were evaluated for their antiviral activity against influenza virus A/Puerto Rico/8/34 (H1N1) and cytotoxicity in MDCK cell culture. The plant material (30 g) was extracted with methanol (300 mL) at room temperature for 24 h, after which the methanolic extracts were filtered, evaporated, and subsequently lyophilized. Evaluation of the potential antiviral activity was conducted by a viral replication inhibition test. Among these medicinal plants, Tussilago farfara, Brassica juncea, Prunus armeniaca, Astragalus membranaceus, Patrinia villosa, and Citrus unshiu showed marked antiviral activity against influenza virus A/H1N1 at concentrations ranging from 0.15625 mg/mL to 1.25 mg/mL, 0.3125 mg/mL to 10 mg/mL, 5 mg/mL to 10 mg/mL, 0.625 mg/mL to 10 mg/mL, 0.625 mg/mL to 10 mg/mL, and 0.3125 mg/mL to 5 mg/mL, respectively. The extracts of Tussilago farfara showed cytotoxicity at concentrations greater than 2.5 mg/mL, whereas the other five main extracts showed no cytotoxicity at concentrations of 10 mg/mL. Taken together, the present results indicated that methanolic extracts of the six main plants might be useful for the treatment of influenza virus H1N1.

Present Status and Future Management Strategies for Sugarcane Yellow Leaf Virus: A Major Constraint to the Global Sugarcane Production

  • Holkar, Somnath Kadappa;Balasubramaniam, Parameswari;Kumar, Atul;Kadirvel, Nithya;Shingote, Prashant Raghunath;Chhabra, Manohar Lal;Kumar, Shubham;Kumar, Praveen;Viswanathan, Rasappa;Jain, Rakesh Kumar;Pathak, Ashwini Dutt
    • The Plant Pathology Journal
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    • v.36 no.6
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    • pp.536-557
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    • 2020
  • Sugarcane yellow leaf virus (SCYLV) is a distinct member of the Polerovirus genus of the Luteoviridae family. SCYLV is the major limitation to sugarcane production worldwide and presently occurring in most of the sugarcane growing countries. SCYLV having high genetic diversity within the species and presently ten genotypes are known to occur based on the complete genome sequence information. SCYLV is present in almost all the states of India where sugarcane is grown. Virion comprises of 180 coat protein units and are 24-29 nm in diameter. The genome of SCYLV is a monopartite and comprised of single-stranded (ss) positive-sense (+) linear RNA of about 6 kb in size. Virus genome consists of six open reading frames (ORFs) that are expressed by sub-genomic RNAs. The SCYLV is phloem-limited and transmitted by sugarcane aphid Melanaphis sacchari in a circulative and non-propagative manner. The other aphid species namely, Ceratovacuna lanigera, Rhopalosiphum rufiabdominalis, and R. maidis also been reported to transmit the virus. The virus is not transmitted mechanically, therefore, its transmission by M. sacchari has been studied in different countries. SCYLV has a limited natural host range and mainly infect sugarcane (Sachharum hybrid), grain sorghum (Sorghum bicolor), and Columbus grass (Sorghum almum). Recent insights in the protein-protein interactions of Polerovirus through protein interaction reporter (PIR) technology enable us to understand viral encoded proteins during virus replication, assembly, plant defence mechanism, short and long-distance travel of the virus. This review presents the recent understandings on virus biology, diagnosis, genetic diversity, virus-vector and host-virus interactions and conventional and next generation management approaches.