8 results on '"Chen, Young-Mao"'
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2. Characterization of betanodavirus quasispecies influences on the subcellular localization and expression of tumor necrosis factor (TNF).
- Author
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Chen YM, Tan CS, Wang TY, Hwong CL, and Chen TY
- Subjects
- Animals, Capsid Proteins immunology, Fish Proteins immunology, RNA Virus Infections veterinary, RNA Virus Infections virology, Tumor Necrosis Factor-alpha immunology, Bass, Capsid Proteins genetics, Fish Diseases immunology, Fish Proteins genetics, Nodaviridae genetics, Quasispecies physiology, Tumor Necrosis Factor-alpha genetics
- Abstract
The aim of this study was to investigate the influence of variant coat proteins (CPs) from different quasispecies of betanodavirus on diverse aspects of nodavirus-induced pathogenesis. It is known that variant CPs can acquire either nuclear or cytoplasmic localization, depending on the nodavirus CP genotype, and this variation may arise during viral replication and influence the regulation of host and viral gene transcription. To investigate the role of these variant CPs in pathogenesis, six variant CP expression plasmids were constructed, each containing different quasispecies CP variants from nodavirus genotype red spotted grouper nervous necrosis virus (RGNNV). The CP expression plasmids were transiently transfected into grouper GF-1 cells. At different times, the cell cycle and cell proliferation were assayed using flow cytometry and methyl thiazolyl tetrazolium (MTT) assays, respectively. The proportion of G
2 /M-phase GF-1 cells transfected with CP expression plasmids was higher than that of cells transfected with the blank plasmid, especially in regards to quasispecies 2 (QS2). The proliferation ratio of cells transfected with the CP expression plasmids was significantly higher than that of cells transfected with the blank plasmid, with the exception of QS6. We also found that the different quasispecies CPs downregulated the promoter activity of the tumor necrosis factor (TNF) gene to different degrees. In addition, this is the first report showing the betanodavirus CP derived from different quasispecies of RGNNV provide evidence of a chronically nodavirus-infected grouper. Overall, this study represents the first comprehensive analysis of variant CPs from grouper with persistent nodavirus infections and their effects on different aspects of pathogenesis., (Copyright © 2020 Elsevier Ltd. All rights reserved.)- Published
- 2020
- Full Text
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3. Cloning and characterisation of type I interferon receptor 1 in orange-spotted grouper (Epinephelus coioides) for response to nodavirus infection.
- Author
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Tang ZZ, Wang TY, Chen YM, and Chen TY
- Subjects
- Amino Acid Sequence, Animals, Base Sequence, Cytokines metabolism, Fish Diseases virology, Fish Proteins chemistry, Fish Proteins genetics, Fish Proteins immunology, Gene Expression Profiling veterinary, Lipopolysaccharides administration & dosage, Nodaviridae physiology, Phylogeny, Poly I-C administration & dosage, RNA Virus Infections immunology, RNA Virus Infections veterinary, RNA Virus Infections virology, Receptor, Interferon alpha-beta chemistry, Sequence Alignment veterinary, Bass genetics, Bass immunology, Fish Diseases immunology, Gene Expression Regulation immunology, Immunity, Innate genetics, Receptor, Interferon alpha-beta genetics, Receptor, Interferon alpha-beta immunology
- Abstract
Grouper is known as a highly economical teleost species in the Asian aquaculture industry; however, intensive culture activities easily cause disease outbreak, especially viral disease. For the prevention of viral outbreaks, interferon (IFN) is among the major defence systems being studied in different species. Fish type I IFNs are known to possess antiviral properties similar to mammalian type I IFNs. In order to stimulate antiviral function, IFN will bind to its cognate receptor, the type I interferon receptor (IFNAR), composed of heterodimeric receptor subunits known as IFNAR1 and IFNΑR2. The binding of type I interferon to receptors assists in the transduction of signals from the external to internal environments of cells to activate biological responses. In order to study the function of IFN, we first need to understand IFN receptors. In this study, we cloned and identified IFNAR1 in orange-spotted grouper (osgIFNAR1) and noted the up-regulated mRNA expression of the receptor and downstream effectors in the head kidney cells with cytokine treatment. The transcriptional expression of osgIFNAR1, which is characterised using polyinosinic-polycytidylic acid (poly[I:C]) and lipopolysaccharide (LPS) treatments, indicated the involvement of osgIFNAR1 in the immune response of grouper. The subcellular localisation of osgIFNAR1 demonstrated scattering across the grouper cell. Viral infection showed the negative feedback regulation of osgIFNAR1 in grouper larvae. Further loss of function of IFNAR1 showed a decreased expression of the virus. This study reported the identification of osgIFNAR1 and characterisation of receptor sensitivity towards immunostimulants, cytokine response, and viral challenge in the interferon pathway of orange-spotted grouper and possible different role of the receptor in viral production. Together, these results provide a frontline report of the potential function of osgIFNAR1 in the innate immunity of teleost., (Copyright © 2020 Elsevier Ltd. All rights reserved.)
- Published
- 2020
- Full Text
- View/download PDF
4. Molecular cloning of orange-spotted grouper (Epinephelus coioides) heat shock transcription factor 1 isoforms and characterization of their expressions in response to nodavirus.
- Author
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Wang TY, Chen YM, and Chen TY
- Subjects
- Amino Acid Sequence, Animals, Base Sequence, Cloning, Molecular, DNA, Complementary genetics, DNA, Complementary metabolism, DNA-Binding Proteins chemistry, DNA-Binding Proteins metabolism, Fish Proteins chemistry, Fish Proteins metabolism, Heat Shock Transcription Factors, Hot Temperature adverse effects, Immunity drug effects, Lipopolysaccharides pharmacology, Nodaviridae physiology, Phylogeny, Poly I-C pharmacology, Protein Isoforms chemistry, Protein Isoforms genetics, Protein Isoforms metabolism, RNA Virus Infections immunology, Sequence Alignment veterinary, Transcription Factors chemistry, Transcription Factors metabolism, Bass, DNA-Binding Proteins genetics, Fish Diseases immunology, Fish Proteins genetics, Gene Expression Regulation immunology, RNA Virus Infections veterinary, Transcription Factors genetics
- Abstract
Heat shock transcription factor 1 (HSF1) regulates heat shock proteins (HSPs), which assist in protein folding and inhibit protein denaturation following stress. HSF1 was firstly cloned from orange-spotted grouper and exists as two isoforms, one with (osgHSF1a) and one without (osgHSF1b) exon 11. Heat exposure increased the expression of osgHSF1b while cold exposure increased that of osgHSF1a. Both isoforms were mainly expressed in the brains, eyes, and fins. Expression of osgHSF1b was higher than osgHSF1a during development. Poly I:C and LPS could also induce osgHSF1 isoforms expression differentially. Exposure to nervous necrosis virus (NNV) increased the level of both osgHSF1 isoforms at 12 h. GF-1 cells with overexpression of osgHSF1 isoforms enhanced viral loads within 24 h, whereas both pharmacological inhibition and RNA interference of HSF1 reduced virus infection. This study shows that osgHSF1 can support the early stage of virus infection and provides a new insight into the molecular regulation of osgHSF1 between the influence of temperatures and immunity., (Copyright © 2016 Elsevier Ltd. All rights reserved.)
- Published
- 2016
- Full Text
- View/download PDF
5. Molecular cloning and characterization of orange-spotted grouper (Epinephelus coioides) CXC chemokine ligand 12.
- Author
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Wu CS, Wang TY, Liu CF, Lin HP, Chen YM, and Chen TY
- Subjects
- Amino Acid Sequence, Animals, Base Sequence, Chemokines, CXC metabolism, Cloning, Molecular, DNA, Complementary genetics, DNA, Complementary metabolism, Embryo, Nonmammalian immunology, Fish Diseases virology, Fish Proteins chemistry, Fish Proteins metabolism, Gene Expression Regulation, Developmental, Ligands, Molecular Sequence Data, Nodaviridae physiology, Phylogeny, RNA Virus Infections immunology, RNA Virus Infections virology, Real-Time Polymerase Chain Reaction veterinary, Sequence Alignment veterinary, Tissue Distribution, Chemokines, CXC chemistry, Chemokines, CXC genetics, Fish Diseases immunology, Fish Proteins genetics, Immunity, Innate, Perciformes, RNA Virus Infections veterinary
- Abstract
Chemokines are a family of soluble peptides that can recruit a wide range of immune cells to sites of infection and disease. The CXCL12 is a chemokine that binds to its cognate receptor CXCR4 and thus involved in multiple physiological and pathophysiological processes. In this study, we cloned and characterized CXCL12 from Epinephelus coioides (osgCXCL12). We found that the open reading frame of osgCXCL12 consists of 98 amino acid residues with the small cytokine C-X-C domain located between residues 29 and 87. Higher expression levels for osgCXCL12 were detected at the kitting stage, compared with the prolarva and larva shape stages. The expression patterns revealed that osgCXCL12 may play a key role in early grouper development. We detected mRNA transcripts for osgCXCL12 in healthy tissues and found the highest osgCXCL12 expression in the head kidney. Furthermore, a time-course analysis revealed significantly increased osgCXCL12 and osgCXCR4 expression levels after the nervous necrosis virus (NNV) challenge. In addition, expression of osgCXCL12 was affected by injection with microbial mimics [LPS and poly(I:C)]. These results suggest that osgCXCL12 is associated with inflammatory and developmental processes in the grouper., (Copyright © 2015 Elsevier Ltd. All rights reserved.)
- Published
- 2015
- Full Text
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6. Molecular cloning and functional analysis of an orange-spotted grouper (Epinephelus coioides) secreted protein acidic and rich in cysteine (SPARC) and characterization of its expression response to nodavirus.
- Author
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Chen YM, Kuo CE, Huang YL, Shie PS, Liao JJ, Yang YC, and Chen TY
- Subjects
- Amino Acid Sequence, Animals, Base Sequence, Bass genetics, Cloning, Molecular, DNA, Complementary genetics, DNA, Complementary immunology, Fish Proteins chemistry, Fish Proteins genetics, Fish Proteins immunology, Gene Expression Regulation, Molecular Sequence Data, Osteonectin chemistry, Osteonectin genetics, RNA Virus Infections veterinary, Sequence Alignment, Bass immunology, Fish Diseases immunology, Nodaviridae, Osteonectin immunology, RNA Virus Infections immunology
- Abstract
Mammalian secreted protein acidic and rich in cysteine (SPARC) is the primary regulator of cell shape and cell adhesion to fibronectin. We, for the first time, report the complete sequencing of SPARC cDNA from orange-spotted grouper. Despite the difference in the lengths of the SPARC transcripts, all of the SPARC molecules encoded a signal peptide, follistain-like copper binding sequence (KGHK) domain, and extracellular domain. The grouper SPARC gene was differentially expressed in vivo and contributed differently to high-level expression of SPARC in muscle. Immunohistochemical staining demonstrated a decreased level of SPARC in nodavirus-infected grouper compared with healthy grouper. Comparative real-time polymerase chain reaction analyses of eye tissues of viral nervous necrosis grouper and healthy grouper were performed. Recombinant SPARC produced changes in grouper cell shape 24 h after treatment. The results provide new insight into the pathogenesis of nodavirus, and demonstrate an experimental rationale for SPARC characterization in nodavirus-infected grouper., (Copyright © 2011 Elsevier Ltd. All rights reserved.)
- Published
- 2011
- Full Text
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7. Cloning of an orange-spotted grouper Epinephelus coioides heat shock protein 90AB (HSP90AB) and characterization of its expression in response to nodavirus.
- Author
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Chen YM, Kuo CE, Wang TY, Shie PS, Wang WC, Huang SL, Tsai TJ, Chen PP, Chen JC, and Chen TY
- Subjects
- Amino Acid Sequence, Animals, Antibiotics, Antineoplastic pharmacology, Base Sequence, Benzoquinones pharmacology, Capsid Proteins metabolism, Cell Line, Cloning, Molecular, Gene Expression Profiling, Gene Expression Regulation drug effects, Lactams, Macrocyclic pharmacology, Molecular Sequence Data, Nodaviridae, Phylogeny, RNA Virus Infections immunology, Time Factors, Bass genetics, Bass immunology, Fish Diseases immunology, Heat-Shock Proteins genetics, Heat-Shock Proteins immunology, RNA Virus Infections veterinary
- Abstract
The heat shock proteins (HSPs) family which consists of HSP90, HSP70, and low molecular mass HSPs are involved in chaperone activity. Here, we report the cloning and characterization of HSP90AB gene from orange-spotted grouper, Epinephelus coioides. The full-length of grouper HSP90AB was 727 amino acids and possessed an ATPase domain as well as an evolutionarily conserved molecular chaperone. The HSP90AB-green fluorescent protein fusion protein was evenly distributed in the cytoplasm. Immunohistochemistry (IHC) and real-time polymerase chain reaction (PCR) analyses indicated that the expression of grouper HSP90AB was marginally increased following nodavirus infection. Grouper E. coioides that received HSP90 inhibitor geldanamycin (GA) showed an increase in HSP90AB expression and growth of nodavirus supporting nodavirus replication., (2010 Elsevier Ltd. All rights reserved.)
- Published
- 2010
- Full Text
- View/download PDF
8. Cloning of an orange-spotted grouper (Epinephelus coioides) Mx cDNA and characterisation of its expression in response to nodavirus.
- Author
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Chen YM, Su YL, Lin JH, Yang HL, and Chen TY
- Subjects
- Amino Acid Sequence, Animals, Base Sequence, Cloning, Molecular, Cluster Analysis, DNA Primers, Green Fluorescent Proteins, Molecular Sequence Data, Myxovirus Resistance Proteins, Phylogeny, Poly I-C metabolism, RNA Virus Infections metabolism, Response Elements genetics, Reverse Transcriptase Polymerase Chain Reaction veterinary, Sequence Analysis, DNA veterinary, Transfection veterinary, Fish Diseases metabolism, Fish Diseases virology, GTP-Binding Proteins genetics, Gene Expression Regulation, Nodaviridae, Perciformes genetics, RNA Virus Infections veterinary
- Abstract
Molecular cloning and nucleotide sequencing of cDNA encoding an orange-spotted grouper (Epinephelus coioides) homolog of Mx ("OsgMx") was conducted and its possible role in fish immunity was analysed. Similar to mammalian Mx, the OsgMx are members of a family of interferon-inducible genes that are expressed by cells in response to nodavirus and iridovirus naturally-infected. Expression of OsgMx mRNA was noticeably upregulated in all tissues by nodavirus naturally-infected grouper. The transcription of OsgMx gene increased 6 h after intramuscular injection of nodavirus experimentally-infected fish and peaked at 72 h in their brains. Analysis of the 5'-flanking sequence of the gene shows that as in pufferfish and zebrafish, the OsgMx promoter contains two potential interferon-stimulated response element (ISRE) responsible for the induction of interferon-inducer polyinosinic-polycytidylic acid (Poly[I:C]). Transient transfection of grouper cells in gfp-reporter gene assays shows that the activation of the grouper Mx promoter fragment by Poly[I:C] is sufficient to allow the expression of green fluorescent protein (GFP). These results may provide a possible regulated pathway against nodavirus.
- Published
- 2006
- Full Text
- View/download PDF
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