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Sudsat P, Srisala J, Pakotiprapha D, Tapaneeyakorn S, Sritunyalucksana K, Thitamadee S, Charoensutthivarakul S, Itsathitphaisarn O. VP28 interacts with PmRab7 irrespective of its nucleotide state. Sci Rep 2024; 14:27803. [PMID: 39537865 PMCID: PMC11560936 DOI: 10.1038/s41598-024-79310-5] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Academic Contribution Register] [Received: 05/28/2024] [Accepted: 11/07/2024] [Indexed: 11/16/2024] Open
Abstract
In shrimp aquaculture, white spot syndrome virus (WSSV) infections severely impact production. Previous research highlighted the crucial role of the Penaeus monodon Rab7 (PmRab7) protein in WSSV entry, specifically its interaction with the viral envelope protein VP28. PmRab7 exists in two conformations: GDP-bound (inactive) and GTP-bound (active). This study, using ELISA and isothermal titration calorimetry (ITC), reveals that the PmRab7-VP28 interaction occurs irrespective of the nucleotide binding state of PmRab7. Comparing the binding affinity between VP28 and different PmRab7 conformations, including wild-type (WT, 22.5 nM), a fast nucleotide exchange (L129F, 128 nM), a GDP-bound form (T22N, 334 nM), and a favorably GTP-bound form (Q67L, 1990 nM), PmRab7-WT exhibits the strongest binding affinity, especially at a lower temperature (25 °C). The binding of PmRab7-WT and VP28 in the presence of excess nucleotide (WT with excess GDP, 924 nM, and WT with excess GTP, 826 nM) shows a 2-fold higher binding affinity than in the absence (WT, 1920 nM) indicating that the addition of excess nucleotide for PmRab7-WT enhanced the affinity for VP28. Together, these findings support the potential of PmRab7-WT as a promising therapeutic candidate for WSSV control in shrimp. Furthermore, from an industrial point of view, the ITC platform developed to study the VP28-PmRab7 interactions provides a high-throughput method for screening additives for shrimp feed that can inhibit this interaction.
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Affiliation(s)
- Patcha Sudsat
- Department of Biochemistry, Faculty of Science, Mahidol University, Rama VI Rd, Bangkok, 10400, Thailand
| | - Jiraporn Srisala
- National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), 113 Thailand Science Park, Phahonyothin Rd., Klong Neung, Klong Luang, Pathum Thani, 12120, Thailand
| | - Danaya Pakotiprapha
- Department of Biochemistry, Faculty of Science, Mahidol University, Rama VI Rd, Bangkok, 10400, Thailand
- Center for Excellence in Protein and Enzyme Technology (CPET), Faculty of Science, Mahidol University, Bangkok, 10400, Thailand
| | - Satita Tapaneeyakorn
- National Nanotechnology Center (NANOTEC), National Science and Technology Development Agency (NSTDA), Thailand Science Park, Pathum Thani, 12120, Thailand
| | - Kallaya Sritunyalucksana
- National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), 113 Thailand Science Park, Phahonyothin Rd., Klong Neung, Klong Luang, Pathum Thani, 12120, Thailand
- Center of Excellence in Shrimp Molecular Biology and Biotechnology (Centex Shrimp), Faculty of Science, Mahidol University, Bangkok, 10400, Thailand
| | - Siripong Thitamadee
- Center of Excellence in Shrimp Molecular Biology and Biotechnology (Centex Shrimp), Faculty of Science, Mahidol University, Bangkok, 10400, Thailand
- Department of Biotechnology, Faculty of Science, Mahidol University, Rama VI Rd, Bangkok, 10400, Thailand
- Analytical Sciences and National Doping Test Institute, Mahidol University, Bangkok, 10400, Thailand
| | - Sitthivut Charoensutthivarakul
- School of Bioinnovation and Bio-based Product Intelligence, Faculty of Science, Mahidol University, Bangkok, 10400, Thailand
- Excellent Center for Drug Discovery (ECDD), Faculty of Science, Mahidol University, Bangkok, 10400, Thailand
| | - Ornchuma Itsathitphaisarn
- Department of Biochemistry, Faculty of Science, Mahidol University, Rama VI Rd, Bangkok, 10400, Thailand.
- Center of Excellence in Shrimp Molecular Biology and Biotechnology (Centex Shrimp), Faculty of Science, Mahidol University, Bangkok, 10400, Thailand.
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Boštjančić LL, Francesconi C, Rutz C, Hoffbeck L, Poidevin L, Kress A, Jussila J, Makkonen J, Feldmeyer B, Bálint M, Schwenk K, Lecompte O, Theissinger K. Host-pathogen coevolution drives innate immune response to Aphanomyces astaci infection in freshwater crayfish: transcriptomic evidence. BMC Genomics 2022; 23:600. [PMID: 35989333 PMCID: PMC9394032 DOI: 10.1186/s12864-022-08571-z] [Citation(s) in RCA: 8] [Impact Index Per Article: 2.7] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Academic Contribution Register] [Received: 11/08/2021] [Accepted: 04/20/2022] [Indexed: 02/08/2023] Open
Abstract
BACKGROUND For over a century, scientists have studied host-pathogen interactions between the crayfish plague disease agent Aphanomyces astaci and freshwater crayfish. It has been hypothesised that North American crayfish hosts are disease-resistant due to the long-lasting coevolution with the pathogen. Similarly, the increasing number of latent infections reported in the historically sensitive European crayfish hosts seems to indicate that similar coevolutionary processes are occurring between European crayfish and A. astaci. Our current understanding of these host-pathogen interactions is largely focused on the innate immunity processes in the crayfish haemolymph and cuticle, but the molecular basis of the observed disease-resistance and susceptibility remain unclear. To understand how coevolution is shaping the host's molecular response to the pathogen, susceptible native European noble crayfish and invasive disease-resistant marbled crayfish were challenged with two A. astaci strains of different origin: a haplogroup A strain (introduced to Europe at least 50 years ago, low virulence) and a haplogroup B strain (signal crayfish in lake Tahoe, USA, high virulence). Here, we compare the gene expression profiles of the hepatopancreas, an integrated organ of crayfish immunity and metabolism. RESULTS We characterised several novel innate immune-related gene groups in both crayfish species. Across all challenge groups, we detected 412 differentially expressed genes (DEGs) in the noble crayfish, and 257 DEGs in the marbled crayfish. In the noble crayfish, a clear immune response was detected to the haplogroup B strain, but not to the haplogroup A strain. In contrast, in the marbled crayfish we detected an immune response to the haplogroup A strain, but not to the haplogroup B strain. CONCLUSIONS We highlight the hepatopancreas as an important hub for the synthesis of immune molecules in the response to A. astaci. A clear distinction between the innate immune response in the marbled crayfish and the noble crayfish is the capability of the marbled crayfish to mobilise a higher variety of innate immune response effectors. With this study we outline that the type and strength of the host immune response to the pathogen is strongly influenced by the coevolutionary history of the crayfish with specific A. astaci strains.
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Affiliation(s)
- Ljudevit Luka Boštjančić
- LOEWE Centre for Translational Biodiversity Genomics (LOEWE-TBG), Senckenberg Biodiversity and Climate Research Centre, Georg-Voigt-Str. 14-16, 60325, Frankfurt am Main, Germany
| | - Caterina Francesconi
- Institute for Environmental Sciences, University of Koblenz-Landau, Fortstrasse 7, 76829, Landau, Germany.
| | - Christelle Rutz
- Department of Computer Science, ICube, UMR 7357, University of Strasbourg, CNRS, Centre de Recherche en Biomédecine de Strasbourg, Rue Eugène Boeckel 1, 67000, Strasbourg, France
| | - Lucien Hoffbeck
- Department of Computer Science, ICube, UMR 7357, University of Strasbourg, CNRS, Centre de Recherche en Biomédecine de Strasbourg, Rue Eugène Boeckel 1, 67000, Strasbourg, France
| | - Laetitia Poidevin
- Department of Computer Science, ICube, UMR 7357, University of Strasbourg, CNRS, Centre de Recherche en Biomédecine de Strasbourg, Rue Eugène Boeckel 1, 67000, Strasbourg, France
| | - Arnaud Kress
- Department of Computer Science, ICube, UMR 7357, University of Strasbourg, CNRS, Centre de Recherche en Biomédecine de Strasbourg, Rue Eugène Boeckel 1, 67000, Strasbourg, France
| | - Japo Jussila
- Department of Environmental and Biological Sciences, University of Eastern Finland, P.O. Box 1627, 70210, Kuopio, Finland
| | - Jenny Makkonen
- Department of Environmental and Biological Sciences, University of Eastern Finland, P.O. Box 1627, 70210, Kuopio, Finland
- Present address: BioSafe - Biological Safety Solutions, Microkatu 1, 70210, Kuopio, Finland
| | - Barbara Feldmeyer
- LOEWE Centre for Translational Biodiversity Genomics (LOEWE-TBG), Senckenberg Biodiversity and Climate Research Centre, Georg-Voigt-Str. 14-16, 60325, Frankfurt am Main, Germany
| | - Miklós Bálint
- LOEWE Centre for Translational Biodiversity Genomics (LOEWE-TBG), Senckenberg Biodiversity and Climate Research Centre, Georg-Voigt-Str. 14-16, 60325, Frankfurt am Main, Germany
| | - Klaus Schwenk
- Institute for Environmental Sciences, University of Koblenz-Landau, Fortstrasse 7, 76829, Landau, Germany
| | - Odile Lecompte
- Department of Computer Science, ICube, UMR 7357, University of Strasbourg, CNRS, Centre de Recherche en Biomédecine de Strasbourg, Rue Eugène Boeckel 1, 67000, Strasbourg, France
| | - Kathrin Theissinger
- LOEWE Centre for Translational Biodiversity Genomics (LOEWE-TBG), Senckenberg Biodiversity and Climate Research Centre, Georg-Voigt-Str. 14-16, 60325, Frankfurt am Main, Germany
- Institute for Environmental Sciences, University of Koblenz-Landau, Fortstrasse 7, 76829, Landau, Germany
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Cui C, He L, Tang X, Xing J, Sheng X, Chi H, Zhan W. Monoclonal antibodies (mAbs) and single chain variable fragment (scFv) antibodies targeting envelope protein VP28 of white spot syndrome virus provide protection against viral infection. FISH & SHELLFISH IMMUNOLOGY 2022; 127:508-520. [PMID: 35768048 DOI: 10.1016/j.fsi.2022.06.042] [Citation(s) in RCA: 3] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Academic Contribution Register] [Received: 06/07/2022] [Revised: 06/22/2022] [Accepted: 06/23/2022] [Indexed: 06/15/2023]
Abstract
White spot syndrome virus (WSSV) is extremely pathogenic and causes huge economic losses in the shrimp farming industry. Neutralizing antibodies against WSSV is expected to be an effective means of preventing infection with the virus. In the present study, eight monoclonal antibodies (mAbs) against VP28 were developed by immunizing BALB/c mice with WSSV-VP28 recombinant protein. Among them, three mAbs named 3B7, 2G3 and 5D2 were determined to be able to delay the mortality of WSSV-infected shrimp in vivo neutralization assay, suggesting their neutralizing ability against WSSV infection. Immunoblotting results showed that the three mAbs reacted specifically with native VP28 of WSSV, and could also recognize the virions in the gills of WSSV-infected shrimp by IFA. Furthermore, the single chain variable fragment (scFv) genes specific for WSSV-VP28 were cloned from the three hybridoma cells and expressed in Escherichia coli. After purification and refolding, three biologically active scFv recombinant proteins were all capable of recognizing the native VP28 of WSSV and delayed the mortality of WSSV-infected shrimp, indicating their neutralizing capacity against WSSV. Subsequently, the eukaryotic expression plasmids of three scFv genes were constructed and the transcriptional properties of expression vectors in shrimp were analyzed. Animal experiments also proved that the scFv eukaryotic expression plasmids were able to partially neutralize WSSV infection. Thus, the production of neutralizing mAb and recombinant scFv antibodies against WSSV has a promising therapeutic potential in prevention and treatment of white spot disease of shrimp.
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Affiliation(s)
- Chuang Cui
- Laboratory of Pathology and Immunology of Aquatic Animals, KLMME, Ocean University of China, Qingdao, 266003, China
| | - Liangyin He
- Laboratory of Pathology and Immunology of Aquatic Animals, KLMME, Ocean University of China, Qingdao, 266003, China
| | - Xiaoqian Tang
- Laboratory of Pathology and Immunology of Aquatic Animals, KLMME, Ocean University of China, Qingdao, 266003, China; Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao, 266071, China.
| | - Jing Xing
- Laboratory of Pathology and Immunology of Aquatic Animals, KLMME, Ocean University of China, Qingdao, 266003, China; Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao, 266071, China
| | - Xiuzhen Sheng
- Laboratory of Pathology and Immunology of Aquatic Animals, KLMME, Ocean University of China, Qingdao, 266003, China
| | - Heng Chi
- Laboratory of Pathology and Immunology of Aquatic Animals, KLMME, Ocean University of China, Qingdao, 266003, China; Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao, 266071, China
| | - Wenbin Zhan
- Laboratory of Pathology and Immunology of Aquatic Animals, KLMME, Ocean University of China, Qingdao, 266003, China; Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao, 266071, China
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Zenke K, Okinaka Y. Multiple isoforms of HSP70 and HSP90 required for betanodavirus multiplication in medaka cells. Arch Virol 2022; 167:1961-1975. [PMID: 35752988 DOI: 10.1007/s00705-022-05489-5] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Academic Contribution Register] [Received: 02/28/2022] [Accepted: 05/18/2022] [Indexed: 11/28/2022]
Abstract
Heat shock proteins (HSPs) are molecular chaperones that have recently been shown to function as host factors (HFs) for virus multiplication in fish as well as in mammals, plants, and insects. HSPs are classified into families, and each family has multiple isoforms. However, no comprehensive studies have been performed to clarify the biological importance of these multiple isoforms for fish virus multiplication. Betanodaviruses are the causative agents of viral nervous necrosis in cultured marine fish and cause very high mortality. Although the viral genome and encoded proteins have been characterized extensively, information on HFs for these viruses is limited. In this study, therefore, we focused on the HSP70 and HSP90 families to examine the importance of their isoforms for betanodavirus multiplication. We found that HSP inhibitors (17-AAG, radicicol, and quercetin) suppressed viral RNA replication and production of progeny virus in infected medaka (Oryzias latipes) cells. Thermal stress or virus infection resulted in increased expression of some isoform genes and facilitated virus multiplication. Furthermore, overexpression and knockdown of some isoform genes revealed that the isoforms HSP70-1, HSP70-2, HSP70-5, HSP90-α1, HSP90-α2, and HSP90-β play positive roles in virus multiplication in medaka. Collectively, these results suggest that multiple isoforms of fish HPSs serve as HFs for betanodavirus multiplication.
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Affiliation(s)
- Kosuke Zenke
- Graduate School of Integrated Sciences for Life, Hiroshima University, Higashi-Hiroshima, 739-8528, Japan.,Research Institute of Pharmaceutical Sciences, Musashino University, 1-1-20 Shinmachi, Nishitokyo-shi, Tokyo, 202-8585, Japan
| | - Yasushi Okinaka
- Graduate School of Integrated Sciences for Life, Hiroshima University, Higashi-Hiroshima, 739-8528, Japan.
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In silico studies on the interaction of phage displayed biorecognition element (TFQAFDLSPFPS) with the structural protein VP28 of white spot syndrome virus. J Mol Model 2020; 26:264. [PMID: 32914310 DOI: 10.1007/s00894-020-04524-z] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Academic Contribution Register] [Received: 04/07/2020] [Accepted: 08/26/2020] [Indexed: 01/21/2023]
Abstract
White spot disease caused by the white spot syndrome virus (WSSV) incurs a huge loss to the shrimp farming industry. Since no effective therapeutic measures are available, early detection and prevention of the disease are indispensable. Towards this goal, we previously identified a 12-mer phage displayed peptide (designated as pep28) with high affinity for VP28, the structural protein of the white spot syndrome virus (WSSV). The peptide pep28 was successfully used as a biorecognition probe in the lateral flow assay developed for rapid, on-site detection of WSSV. To unravel the structural determinants for the selective binding between VP28 and pep28, we used bioinformatics, structural modeling, protein-protein docking, and binding-free energy studies. We performed atomistic molecular dynamics simulations of pep28-pIII model totaling 300 ns timescale. The most representative pep28-pIII structure from the simulation was used for docking with the crystal structure of VP28. Our results reveal that pep28 binds in a surface groove of the monomeric VP28 β-barrel and makes several hydrogen bonds and non-polar interactions. Ensemble-based binding-free energy studies reveal that the binding is dominated by non-polar interactions. Our studies provide molecular level insights into the binding mechanism of pep28 with VP28, which explain why the peptide is selective and can assist in modifying pep28 for its practical use, both as a biorecognition probe and a therapeutic.
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Lu Y, Qiu Q, Li C, Cheng L, Liu J. Computational identification of self-inhibitory peptides from white spot syndrome virus envelope protein VP28. AQUACULTURE REPORTS 2019; 14:100195. [PMID: 32289063 PMCID: PMC7104047 DOI: 10.1016/j.aqrep.2019.100195] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Subscribe] [Academic Contribution Register] [Received: 01/14/2019] [Revised: 03/31/2019] [Accepted: 04/08/2019] [Indexed: 06/11/2023]
Abstract
Since effective chemotherapeutics or preventive measures are still unavailable, finding feasible approaches against white spot syndrome virus (WSSV) has always been the vital subject in shrimp farming field. Envelope proteins are the ideal targets for antiviral strategies development due to their indispensable roles in virus entry, and inhibitory peptides targeting them have been proved to be promising in blocking virus infection. In this study, the Wimley-White interfacial hydrophobicity scale (WWIHS) in combination with known structural data was applied to identify potential inhibitory peptides that targeted the envelope protein VP28 of WSSV. Results showed that two potential inhibitory peptides were identified, one of which exhibited not only obvious antiviral activity, but also broad-spectrum antimicrobial activity. The inhibitory peptide identified here can serve as a lead compound for anti-WSSV strategies development.
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Affiliation(s)
- Yongzhong Lu
- Shandong Provincial Key Laboratory of Biochemical Engineering, Qingdao University of Science and Technology, No. 53, Zhengzhou Road, 266042, Qingdao, China
| | - Qian Qiu
- Shandong Provincial Key Laboratory of Biochemical Engineering, Qingdao University of Science and Technology, No. 53, Zhengzhou Road, 266042, Qingdao, China
| | - Chen Li
- Yellow Sea Fisheries Research Institute, Chinese Academy Of Fishery Sciences, No. 106, Nanjing Road, 266071, Qingdao, China
| | - Linyue Cheng
- Shandong Provincial Key Laboratory of Biochemical Engineering, Qingdao University of Science and Technology, No. 53, Zhengzhou Road, 266042, Qingdao, China
| | - Jie Liu
- Shandong Provincial Key Laboratory of Biochemical Engineering, Qingdao University of Science and Technology, No. 53, Zhengzhou Road, 266042, Qingdao, China
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7
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Yuan Z, Chen M, Wang J, Li Z, Geng X, Sun J. Identification of Litopenaeus vannamei BiP as a novel cellular attachment protein for white spot syndrome virus by using a biotinylation based affinity chromatography method. FISH & SHELLFISH IMMUNOLOGY 2018; 79:130-139. [PMID: 29738871 DOI: 10.1016/j.fsi.2018.05.003] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Academic Contribution Register] [Received: 01/15/2018] [Revised: 04/09/2018] [Accepted: 05/02/2018] [Indexed: 06/08/2023]
Abstract
White spot syndrome virus (WSSV) is a dangerous threat to shrimp farming that also attacks a wide range of crustaceans. Knowledge of the surface protein-protein interactions between the pathogen and host is very crucial to unraveling the molecular pathogenesis mechanisms of WSSV. In this study, LvBiP (Litopenaeus vannamei immunoglobulin heavy-chain-binding protein) was identified as a novel WSSV binding protein of L. vannamei by a biotinylation based affinity chromatography method. By using pull-down and ELISA assays, the binding of recombinant LvBiP to WSSV was proved to be specific and ATP- dependent. The interaction was also confirmed by the result of co-immunoprecipitation assay. Immunofluorescence studies revealed the co-localization of LvBiP with WSSV on the cell surface of shrimp haemocytes. Additionally, LvBiP is likely to play an important role in WSSV infection. Treatment of gill cellular membrane proteins (CMPs) with purified rLvBiP and antibody that specifically recognizes LvBiP, led to a significant reduction in the binding of WSSV to gill CMPs. In the in vivo neutralization assay, rLvBiP and anti-LvBiP polyclonal antibody partially blocked the infection of WSSV. Taken together, the results indicate that LvBiP, a molecular chaperon of the HSP70 family, is a novel host factor involved at the step of attachment of the WSSV to the host cells and a potential candidate of therapeutic target.
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Affiliation(s)
- Zengzhi Yuan
- Tianjin Key Laboratory of Animal and Plant Resistance, Tianjin, 300387, PR China; College of Life Sciences, Tianjin Normal University, 393 West Binshui Road, Xiqing District, Tianjin, 300387, PR China
| | - Meng Chen
- College of Life Sciences, Tianjin Normal University, 393 West Binshui Road, Xiqing District, Tianjin, 300387, PR China
| | - Jingting Wang
- College of Life Sciences, Tianjin Normal University, 393 West Binshui Road, Xiqing District, Tianjin, 300387, PR China
| | - Zhuoyu Li
- College of Life Sciences, Tianjin Normal University, 393 West Binshui Road, Xiqing District, Tianjin, 300387, PR China
| | - Xuyun Geng
- Tianjin Center for Control and Prevention of Aquatic Animal Infectious Disease, 442 South Jiefang Road, Hexi District, Tianjin 300221, China
| | - Jinsheng Sun
- Tianjin Key Laboratory of Animal and Plant Resistance, Tianjin, 300387, PR China; College of Life Sciences, Tianjin Normal University, 393 West Binshui Road, Xiqing District, Tianjin, 300387, PR China.
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8
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Guerrero CA, Acosta O. Inflammatory and oxidative stress in rotavirus infection. World J Virol 2016; 5:38-62. [PMID: 27175349 PMCID: PMC4861870 DOI: 10.5501/wjv.v5.i2.38] [Citation(s) in RCA: 34] [Impact Index Per Article: 3.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Academic Contribution Register] [Received: 08/12/2015] [Revised: 10/23/2015] [Accepted: 01/29/2016] [Indexed: 02/05/2023] Open
Abstract
Rotaviruses are the single leading cause of life-threatening diarrhea affecting children under 5 years of age. Rotavirus entry into the host cell seems to occur by sequential interactions between virion proteins and various cell surface molecules. The entry mechanisms seem to involve the contribution of cellular molecules having binding, chaperoning and oxido-reducing activities. It appears to be that the receptor usage and tropism of rotaviruses is determined by the species, cell line and rotavirus strain. Rotaviruses have evolved functions which can antagonize the host innate immune response, whereas are able to induce endoplasmic reticulum (ER) stress, oxidative stress and inflammatory signaling. A networking between ER stress, inflammation and oxidative stress is suggested, in which release of calcium from the ER increases the generation of mitochondrial reactive oxygen species (ROS) leading to toxic accumulation of ROS within ER and mitochondria. Sustained ER stress potentially stimulates inflammatory response through unfolded protein response pathways. However, the detailed characterization of the molecular mechanisms underpinning these rotavirus-induced stressful conditions is still lacking. The signaling events triggered by host recognition of virus-associated molecular patterns offers an opportunity for the development of novel therapeutic strategies aimed at interfering with rotavirus infection. The use of N-acetylcysteine, non-steroidal anti-inflammatory drugs and PPARγ agonists to inhibit rotavirus infection opens a new way for treating the rotavirus-induced diarrhea and complementing vaccines.
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Chen LH, Lin SW, Liu KF, Chang CI, Hseu JR, Tsai JM. Comparative proteomic analysis of Litopenaeus vannamei gills after vaccination with two WSSV structural proteins. FISH & SHELLFISH IMMUNOLOGY 2016; 49:306-314. [PMID: 26766180 DOI: 10.1016/j.fsi.2015.12.044] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Academic Contribution Register] [Received: 09/17/2015] [Revised: 12/29/2015] [Accepted: 12/30/2015] [Indexed: 06/05/2023]
Abstract
White spot syndrome virus (WSSV) is one of the most devastating viral pathogens of cultured shrimp worldwide. Recently published papers show the ability of WSSV structural protein VP28 to vaccinate shrimp and raise protection against the virus. This study attempted to identify the joining proteins of the aforementioned shrimp quasi-immune response by proteomic analysis. The other envelope protein, VP36B, was used as the non-protective subunit vaccine control. Shrimp were intramuscularly injected with rVPs or PBS on day 1 and day 4 and then on day 7 their gill tissues were sampled. The two-dimensional electrophoresis (2-DE) patterns of gill proteins between vaccinated and PBS groups were compared and 20 differentially expressed proteins identified by mass spectrometry, some of which were validated in gill and hemocyte tissues using real-time quantitative RT-PCR. Many of identified proteins and their expression levels also linked with the shrimp response during WSSV infection. The list of up-regulated protein spots found exclusively in rVP28-vaccinated shrimp include calreticulin and heat shock protein 70 with chaperone properties, ubiquitin, and others. The two serine proteases, chymotrypsin and trypsin, were significantly increased in shrimp of both vaccinated groups compared to PBS controls. The information presented here should be useful for gaining insight into invertebrate immunity.
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Affiliation(s)
- Li-Hao Chen
- Department of Marine Biotechnology, National Kaohsiung Marine University, Kaohsiung, 81157, Taiwan, ROC
| | - Shi-Wei Lin
- Department of Marine Biotechnology, National Kaohsiung Marine University, Kaohsiung, 81157, Taiwan, ROC
| | - Kuan-Fu Liu
- Tungkang Biotechnology Research Center, Fisheries Research Institute, Council of Agriculture, Pingtung, 92845, Taiwan, ROC
| | - Chin-I Chang
- Aquaculture Division, Fisheries Research Institute, Council of Agriculture, Keelung, 20246, Taiwan, ROC
| | - Jinn-Rong Hseu
- Mariculture Research Center, Fisheries Research Institute, Council of Agriculture, Tainan, 72453, Taiwan, ROC
| | - Jyh-Ming Tsai
- Department of Marine Biotechnology, National Kaohsiung Marine University, Kaohsiung, 81157, Taiwan, ROC.
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Syed Musthaq SK, Kwang J. Reprint of "evolution of specific immunity in shrimp - a vaccination perspective against white spot syndrome virus". DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY 2015; 48:342-353. [PMID: 25083808 DOI: 10.1016/j.dci.2014.07.016] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Subscribe] [Academic Contribution Register] [Received: 01/23/2014] [Revised: 04/11/2014] [Accepted: 04/21/2014] [Indexed: 06/03/2023]
Abstract
Invertebrates lack true adaptive immunity and it solely depends on the primitive immunity called innate immunity. However, various innate immune molecules and mechanisms are identified in shrimp that plays potential role against invading bacterial, fungal and viral pathogens. Perceiving the shrimp innate immune mechanisms will contribute in developing effective vaccine strategies against major shrimp pathogens. Hence this review intends to explore the innate immune molecules of shrimp with suitable experimental evidences together with the evolution of "specific immune priming" of invertebrates. In addition, we have emphasized on the development of an effective vaccine strategy against major shrimp pathogen, white spot syndrome virus (WSSV). The baculovirus displayed rVP28 (Bac-VP28), a major envelope protein of WSSV was utilized to study its vaccine efficacy by oral route. A significant advantage of this baculovirus expression cassette is the use of WSSV-immediate early 1 (ie1) promoter that derived the abundant expression of rVP28 protein at the early stage of the infection in insect cell. The orally vaccinated shrimp with Bac-VP28 transduced successfully in the shrimp cells as well as provided highest survival rate. In support to our vaccine efficacy we analysed Pattern Recognition Proteins (PRPs) β-1,3 glucan lipopolysaccharides (LGBP) and STAT gene profiles in the experimental shrimp. Indeed, the vaccination of shrimp with Bac-VP28 demonstrated some degree of specificity with enhanced survival rate when compared to control vaccination with Bac-wt. Hence it is presumed that the concept of "specific immune priming" in relevant to shrimp immunity is possible but may not be common to all shrimp pathogens.
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Affiliation(s)
- Syed Khader Syed Musthaq
- Animal Health Biotechnology, Temasek Lifesciences Laboratory, 1 Research Link, National University of Singapore, Singapore 117604, Singapore
| | - Jimmy Kwang
- Animal Health Biotechnology, Temasek Lifesciences Laboratory, 1 Research Link, National University of Singapore, Singapore 117604, Singapore; Department of Microbiology, Faculty of Medicine, National University of Singapore, Singapore.
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Valentim-Neto PA, Moser JR, Fraga APM, Marques MRF. Hsp70 expression in shrimp Litopenaeus vannamei in response to IHHNV and WSSV infection. Virusdisease 2014; 25:437-40. [PMID: 25674619 DOI: 10.1007/s13337-014-0236-6] [Citation(s) in RCA: 14] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Academic Contribution Register] [Received: 07/15/2014] [Accepted: 10/08/2014] [Indexed: 10/24/2022] Open
Abstract
The effects of environmental changes, on non-target organisms may impact and bring consequences at the molecular level in marine organisms. There is a lack of data supporting the hypothesis according to which environmental stress modulates the immune system, leading to an increased susceptibility to infectious agents in shrimps. The present study was focused on changes occurring in cellular defense proteins in Litopenaeus vannamei infected by virus infection hypodermal and hematopoietic necrosis (IHHNV) and white spot syndrome (WSSV). Western blot analysis was used to evaluate expression of protein Hsp70, in gill tissue. Up-regulation levels were supported by immune detection analysis, suggesting that IHHNV and WSSV-shrimp infection promotes changes in the expression of these proteins.
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Affiliation(s)
- Pedro A Valentim-Neto
- LABCAI, Dep. Bioquímica, CCB, NEPAq, Universidade Federal de Santa Catarina, Florianópolis, SC 88034-257 Brazil
| | - Juliana R Moser
- LABCAI, Dep. Bioquímica, CCB, NEPAq, Universidade Federal de Santa Catarina, Florianópolis, SC 88034-257 Brazil
| | - Ana P M Fraga
- LABCAI, Dep. Bioquímica, CCB, NEPAq, Universidade Federal de Santa Catarina, Florianópolis, SC 88034-257 Brazil
| | - Maria R F Marques
- LABCAI, Dep. Bioquímica, CCB, NEPAq, Universidade Federal de Santa Catarina, Florianópolis, SC 88034-257 Brazil
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12
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Syed Musthaq SK, Kwang J. Evolution of specific immunity in shrimp - a vaccination perspective against white spot syndrome virus. DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY 2014; 46:279-290. [PMID: 24780624 DOI: 10.1016/j.dci.2014.04.013] [Citation(s) in RCA: 10] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Academic Contribution Register] [Received: 01/23/2014] [Revised: 04/11/2014] [Accepted: 04/21/2014] [Indexed: 06/03/2023]
Abstract
Invertebrates lack true adaptive immunity and it solely depends on the primitive immunity called innate immunity. However, various innate immune molecules and mechanisms are identified in shrimp that plays potential role against invading bacterial, fungal and viral pathogens. Perceiving the shrimp innate immune mechanisms will contribute in developing effective vaccine strategies against major shrimp pathogens. Hence this review intends to explore the innate immune molecules of shrimp with suitable experimental evidences together with the evolution of "specific immune priming" of invertebrates. In addition, we have emphasized on the development of an effective vaccine strategy against major shrimp pathogen, white spot syndrome virus (WSSV). The baculovirus displayed rVP28 (Bac-VP28), a major envelope protein of WSSV was utilized to study its vaccine efficacy by oral route. A significant advantage of this baculovirus expression cassette is the use of WSSV-immediate early 1 (ie1) promoter that derived the abundant expression of rVP28 protein at the early stage of the infection in insect cell. The orally vaccinated shrimp with Bac-VP28 transduced successfully in the shrimp cells as well as provided highest survival rate. In support to our vaccine efficacy we analysed Pattern Recognition Proteins (PRPs) β-1,3 glucan lipopolysaccharides (LGBP) and STAT gene profiles in the experimental shrimp. Indeed, the vaccination of shrimp with Bac-VP28 demonstrated some degree of specificity with enhanced survival rate when compared to control vaccination with Bac-wt. Hence it is presumed that the concept of "specific immune priming" in relevant to shrimp immunity is possible but may not be common to all shrimp pathogens.
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Affiliation(s)
- Syed Khader Syed Musthaq
- Animal Health Biotechnology, Temasek Lifesciences Laboratory, 1 Research Link, National University of Singapore, Singapore 117604, Singapore
| | - Jimmy Kwang
- Animal Health Biotechnology, Temasek Lifesciences Laboratory, 1 Research Link, National University of Singapore, Singapore 117604, Singapore; Department of Microbiology, Faculty of Medicine, National University of Singapore, Singapore.
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Biradar V, Narwade S, Paingankar M, Deobagkar D. White Spot Syndrome Virus infection in Penaeus monodon is facilitated by housekeeping molecules. J Biosci 2014; 38:917-24. [PMID: 24296895 DOI: 10.1007/s12038-013-9386-8] [Citation(s) in RCA: 6] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Academic Contribution Register] [Indexed: 01/26/2023]
Abstract
White Spot Syndrome Virus (WSSV) is a major pathogen in shrimp aquaculture, and its rampant spread has resulted in great economic loss. Identification of host cellular proteins interacting with WSSV will help in unravelling the repertoire of host proteins involved in WSSV infection. In this study, we have employed one-dimensional and two-dimension virus overlay protein binding assay (VOPBA) followed by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) to identify the host proteins of Penaeus monodon that could interact with WSSV. The VOPBA results suggest that WSSV interacted with housekeeping proteins such as heat shock protein 70, ATP synthase subunit beta, phosphopyruvate hydratase, allergen Pen m 2, glyceraldehyde-3-phosphate dehydrogenase, sarcoplasmic calcium-binding protein, actin and 14-3-3-like protein. Our findings suggest that WSSV exploits an array of housekeeping proteins for its transmission and propagation in P. monodon.
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Affiliation(s)
- Vinayak Biradar
- Molecular Biology Research Laboratory, Department of Zoology, Centre for Advance Studies, University of Pune, Pune 411 007, India
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14
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Sun Y, Li F, Sun Z, Zhang X, Li S, Zhang C, Xiang J. Transcriptome analysis of the initial stage of acute WSSV infection caused by temperature change. PLoS One 2014; 9:e90732. [PMID: 24595043 PMCID: PMC3942461 DOI: 10.1371/journal.pone.0090732] [Citation(s) in RCA: 17] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Academic Contribution Register] [Received: 11/16/2013] [Accepted: 02/03/2014] [Indexed: 11/19/2022] Open
Abstract
White spot syndrome virus (WSSV) is the most devastating virosis threatening the shrimp culture industry worldwide. Variations of environmental factors in shrimp culture ponds usually lead to the outbreak of white spot syndrome (WSS). In order to know the molecular mechanisms of WSS outbreak induced by temperature variation and the biological changes of the host at the initial stage of WSSV acute infection, RNA-Seq technology was used to analyze the differentially expressed genes (DEGs) in shrimp with a certain amount of WSSV cultured at 18°C and shrimp whose culture temperature were raised to 25°C. To analyze whether the expression changes of the DEGs were due to temperature rising or WSSV proliferation, the expression of selected DEGs was analyzed by real-time PCR with another shrimp group, namely Group T, as control. Group T didn't suffer WSSV infection but was subjected to temperature rising in parallel. At the initial stage of WSSV acute infection, DEGs related to energy production were up-regulated, whereas most DEGs related to cell cycle and positive regulation of cell death and were down-regulated. Triose phosphate isomerase, enolase and alcohol dehydrogenase involved in glycosis were up-regulated, while pyruvate dehydrogenase, citrate synthase and isocitrate dehydrogenase with NAD as the coenzyme involved in TCA pathway were down-regulated. Also genes involved in host DNA replication, including DNA primase, DNA topoisomerase and DNA polymerase showed down-regulated expression. Several interesting genes including crustin genes, acting binding or inhibiting protein genes, a disintegrin and metalloproteinase domain-containing protein 9 (ADAM9) gene and a GRP 78 gene were also analyzed. Understanding the interactions between hosts and WSSV at the initial stage of acute infection will not only help to get a deep insight into the pathogenesis of WSSV but also provide clues for therapies.
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Affiliation(s)
- Yumiao Sun
- Institute of Oceanology, Chinese Academy of Sciences, Qingdao, China
| | - Fuhua Li
- Institute of Oceanology, Chinese Academy of Sciences, Qingdao, China
| | - Zheng Sun
- Institute of Oceanology, Chinese Academy of Sciences, Qingdao, China
| | - Xiaojun Zhang
- Institute of Oceanology, Chinese Academy of Sciences, Qingdao, China
| | - Shihao Li
- Institute of Oceanology, Chinese Academy of Sciences, Qingdao, China
| | - Chengsong Zhang
- Institute of Oceanology, Chinese Academy of Sciences, Qingdao, China
| | - Jianhai Xiang
- Institute of Oceanology, Chinese Academy of Sciences, Qingdao, China
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15
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Gao H, Lai X, Kong J, Wang W, Meng X, Yan B, Cai S. Cloning of Hsp21 gene and its expression in Chinese shrimp Fenneropenaeus chinensis in response to WSSV challenge. J Appl Genet 2014; 55:231-8. [DOI: 10.1007/s13353-013-0191-8] [Citation(s) in RCA: 13] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Academic Contribution Register] [Received: 04/19/2013] [Revised: 12/09/2013] [Accepted: 12/30/2013] [Indexed: 11/25/2022]
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Zhong R, Tang X, Zhan W, Xing J, Sheng X. Expression kinetics of β-integrin in Chinese shrimp (Fenneropenaeus chinensis) hemocytes following infection with white spot syndrome virus. FISH & SHELLFISH IMMUNOLOGY 2013; 35:539-545. [PMID: 23711468 DOI: 10.1016/j.fsi.2013.05.004] [Citation(s) in RCA: 11] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Academic Contribution Register] [Received: 02/20/2013] [Revised: 04/15/2013] [Accepted: 05/06/2013] [Indexed: 06/02/2023]
Abstract
Our previous study has demonstrated that an integrin β subunit of Chinese shrimp (Fenneropenaeus chinensis) (FcβInt) involved in WSSV infection. In order to further elucidate the potential role of the FcβInt in the WSSV infection, expression response of FcβInt to WSSV infection in shrimp hemocytes was investigated after intra-muscular injection with the virus. Following time-course hemocytes sampling, the expression variation of FcβInt in hemocytes was examined by flow cytometric immunofluorescence assay (FCIFA) and enzyme linked immunosorbent assay (ELISA) using the monoclonal antibody (Mab) 2C5 against FcβInt, which was successfully produced with recombinant partial FcβInt and exhibited binding to a 120 kDa hemocyte protein. Meanwhile, the dynamic state of FcβInt mRNA level and WSSV copies in hemocytes were determined by quantitative real-time PCR. The result of FCIFA showed that FcβInt was mainly expressed on the semi-granular and granular cells, which was down-regulated at 6 h post infection (p.i.), and significantly increased to the peak level at 12 h p.i., then decreased to the control level by 24 h. However, FcβInt on the hyaline cells was lowly expressed and didn't show active response to the viral infection. The variation of FcβInt concentrations in total hemocytes determined by ELISA was roughly in accordance with the changing tendency of FcβInt expressed on the semi-granular and granular cells. FcβInt mRNA level in total hemocytes was significantly up-regulated to the peak level at 12 h p.i. Moreover, the number of WSSV copies in hemocytes began to exhibit a significant increase at 24 h p.i. The present study demonstrated that WSSV infection would induce a differential regulation of FcβInt expression in different type hemocytes, which provided valuable evidences for the close correlation between FcβInt and WSSV infection.
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Affiliation(s)
- Rujie Zhong
- Laboratory of Pathology and Immunology of Aquatic Animals, Ocean University of China, No. 5, Yushan Road, Qingdao 266003, PR China
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17
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Sritunyalucksana K, Utairungsee T, Sirikharin R, Srisala J. Reprint of: Virus-binding proteins and their roles in shrimp innate immunity. FISH & SHELLFISH IMMUNOLOGY 2013; 34:1018-1024. [PMID: 23416697 DOI: 10.1016/j.fsi.2013.02.013] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [Track Full Text] [Subscribe] [Academic Contribution Register] [Received: 06/12/2012] [Revised: 09/01/2012] [Accepted: 09/10/2012] [Indexed: 06/01/2023]
Abstract
Disease outbreaks caused by viral pathogens constitute a major limitation to development of the shrimp aquaculture industry. Many research have been conducted to better understand how host shrimp respond to viral infections with the aim of using the gained knowledge to develop better strategies for disease management and control. One approach has been to study the interactions between host and viral proteins, and particularly host virus-binding proteins that might play an important role in the viral infection process. Within the past five years, increasing numbers of virus-binding proteins (VBPs) have been reported in shrimp. Characterization of these molecules has emphasized on their potential therapeutic applications by demonstrating their activities in inhibition of viral replication via in vivo neutralization assay. However, signaling to induce innate antiviral immune responses as a consequence of binding between viral proteins and VBPs remain to be fully elucidated.
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Affiliation(s)
- Kallaya Sritunyalucksana
- Shrimp-Virus Interaction Laboratory, National Center for Genetic Engineering and Biotechnology (BIOTEC), Thailand Science Park, Klong Luang, Pathumthani 12120, Thailand.
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18
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Sritunyalucksana K, Utairungsee T, Sirikharin R, Srisala J. Virus-binding proteins and their roles in shrimp innate immunity. FISH & SHELLFISH IMMUNOLOGY 2012; 33:1269-1275. [PMID: 23023111 DOI: 10.1016/j.fsi.2012.09.017] [Citation(s) in RCA: 10] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Academic Contribution Register] [Received: 06/12/2012] [Revised: 09/01/2012] [Accepted: 09/10/2012] [Indexed: 06/01/2023]
Abstract
Disease outbreaks caused by viral pathogens constitute a major limitation to development of the shrimp aquaculture industry. Many research have been conducted to better understand how host shrimp respond to viral infections with the aim of using the gained knowledge to develop better strategies for disease management and control. One approach has been to study the interactions between host and viral proteins, and particularly host virus-binding proteins that might play an important role in the viral infection process. Within the past five years, increasing numbers of virus-binding proteins (VBPs) have been reported in shrimp. Characterization of these molecules has emphasized on their potential therapeutic applications by demonstrating their activities in inhibition of viral replication via in vivo neutralization assay. However, signaling to induce innate antiviral immune responses as a consequence of binding between viral proteins and VBPs remain to be fully elucidated.
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Affiliation(s)
- Kallaya Sritunyalucksana
- Shrimp-Virus Interaction Laboratory, National Center for Genetic Engineering and Biotechnology (BIOTEC), Thailand Science Park, Klong Luang, Pathumthani 12120, Thailand.
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Comprehensive review on the HSC70 functions, interactions with related molecules and involvement in clinical diseases and therapeutic potential. Pharmacol Ther 2012; 136:354-74. [PMID: 22960394 DOI: 10.1016/j.pharmthera.2012.08.014] [Citation(s) in RCA: 208] [Impact Index Per Article: 16.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Academic Contribution Register] [Received: 08/14/2012] [Accepted: 08/14/2012] [Indexed: 12/28/2022]
Abstract
Heat shock cognate protein 70 (HSC70) is a constitutively expressed molecular chaperone which belongs to the heat shock protein 70 (HSP70) family. HSC70 shares some of the structural and functional similarity with HSP70. HSC70 also has different properties compared with HSP70 and other heat shock family members. HSC70 performs its full functions by the cooperation of co-chaperones. It interacts with many other molecules as well and regulates various cellular functions. It is also involved in various diseases and may become a biomarker for diagnosis and potential therapeutic targets for design, discovery, and development of novel drugs to treat various diseases. In this article, we provide a comprehensive review on HSC70 from the literatures including the basic general information such as classification, structure and cellular location, genetics and function, as well as its protein association and interaction with other proteins. In addition, we also discussed the relationship of HSC70 and related clinical diseases such as cancer, cardiovascular, neurological, hepatic and many other diseases and possible therapeutic potential and highlight the progress and prospects of research in this field. Understanding the functions of HSC70 and its interaction with other molecules will help us to reveal other novel properties of this protein. Scientists may be able to utilize this protein as a biomarker and therapeutic target to make significant advancement in scientific research and clinical setting in the future.
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20
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Otta SK. Host and virus protein interaction studies in understanding shrimp virus gene function. INDIAN JOURNAL OF VIROLOGY : AN OFFICIAL ORGAN OF INDIAN VIROLOGICAL SOCIETY 2012; 23:184-90. [PMID: 23997442 DOI: 10.1007/s13337-012-0085-0] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Subscribe] [Academic Contribution Register] [Received: 12/31/2011] [Accepted: 06/29/2012] [Indexed: 11/24/2022]
Abstract
Protein-protein interaction studies have been widely used in several fields to characterize an unknown protein. This in turn helps to find out several pathways to understand a complex mechanism or discover a drug for treatment. Among the methods, yeast two-hybrid has widely been used in human, animal and plant research studies. This aspect of research has also been found useful in understanding the shrimp virus gene function. With respect to White spot syndrome virus, interaction studies have been applied to elucidate virus structure, understand the mode of entry of the virus, mechanism of virus replication and also to discover some of the host anti-viral proteins. Interaction studies on other shrimp viruses are scanty and only few reports available on Yellow head virus and Taura syndrome virus. All these findings are still in preliminary stage and lot more studies are necessary to have the clear picture. Protein interaction research on other shrimp viruses are still lacking. Considering all these, it appears that this field of research has a wide scope to understand the virulence mechanism of shrimp viruses where very little information is available till date.
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Affiliation(s)
- Subhendu Kumar Otta
- Aquatic Animal Health Division, Central Institute of Brackishwater Aquaculture, # 75 Santhome High Road, RA Puram, Chennai, 600028 India
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21
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S SM, Kwang J. Oral vaccination of baculovirus-expressed VP28 displays enhanced protection against White Spot Syndrome Virus in Penaeus monodon. PLoS One 2011; 6:e26428. [PMID: 22069450 PMCID: PMC3206036 DOI: 10.1371/journal.pone.0026428] [Citation(s) in RCA: 49] [Impact Index Per Article: 3.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Academic Contribution Register] [Received: 07/29/2011] [Accepted: 09/26/2011] [Indexed: 12/19/2022] Open
Abstract
White Spot Syndrome Virus (WSSV) is an infectious pathogen of shrimp and other crustaceans, and neither effective vaccines nor adequate treatments are currently available. WSSV is an enveloped dsDNA virus, and one of its major envelope proteins, VP28, plays a pivotal role in WSSV infection. In an attempt to develop a vaccine against WSSV, we inserted the VP28 gene into a baculovirus vector tailored to express VP28 on the baculovirus surface under the WSSV ie1 promoter (Bac-VP28). The Bac-VP28 incorporated abundant quantity (65.3 µg/ml) of VP28. Shrimp were treated by oral and immersion vaccination with either Bac-VP28 or wild-type baculovirus (Bac-wt). The treatment was followed by challenge with WSSV after 3 and 15 days. Bac-VP28 vaccinated shrimp showed significantly higher survival rates (oral: 81.7% and 76.7%; immersion: 75% and 68.4%) than Bac-wt or non-treated shrimp (100% mortality). To verify the protective effects of Bac-VP28, we examined in vivo expression of VP28 by immunohistochemistry and quantified the WSSV copy number by qPCR. In addition to that, we quantified the expression levels shrimp genes LGBP and STAT by real-time RT-PCR from the samples obtained from Bac-VP28 vaccinated shrimp at different duration of vaccine regime. Our findings indicate that oral vaccination of shrimp with Bac-VP28 is an attractive preventative measure against WSSV infection that can be used in the field.
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Affiliation(s)
- Syed Musthaq S
- Animal Health Biotechnology, Temasek Lifesciences Laboratory, National University of Singapore, Singapore, Singapore
| | - Jimmy Kwang
- Animal Health Biotechnology, Temasek Lifesciences Laboratory, National University of Singapore, Singapore, Singapore
- Department of Microbiology, Faculty of Medicine, National University of Singapore, Singapore, Singapore
- * E-mail:
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Xiong XP, Dong CF, Xu X, Weng SP, Liu ZY, He JG. Proteomic analysis of zebrafish (Danio rerio) infected with infectious spleen and kidney necrosis virus. DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY 2011; 35:431-440. [PMID: 21075138 DOI: 10.1016/j.dci.2010.11.006] [Citation(s) in RCA: 18] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Academic Contribution Register] [Received: 09/22/2010] [Revised: 11/07/2010] [Accepted: 11/07/2010] [Indexed: 05/30/2023]
Abstract
Iridovirus infections remain a severe problem in aquaculture industries worldwide. Infectious spleen and kidney necrosis virus (ISKNV), the type species of the genus Megalocytovirus in the family Iridoviridae, has caused significant economic losses among freshwater fish in different Asian countries. To investigate the molecular mechanism of iridoviral pathogenesis, we analyzed the differential proteome from the spleen of ISKNV-infected zebrafish through two-dimensional gel electrophoresis (2-DE). Mass spectrometry revealed 35 altered cellular protein spots, including 15 upregulated proteins and 20 downregulated proteins at five days post-infection. The altered host proteins were classified into 13 categories based on their biological processes: cytoskeletal protein, stress response, lipoprotein metabolism, ubiquitin-proteasome pathway, carbohydrate metabolism, signal transduction, proteolysis, ion binding, transport, metabolic process, catabolic process, biosynthesis, and oxidation reduction. Moreover, 14 corresponding genes of the differentially expressed proteins were validated by RT-PCR. Western blot analysis further demonstrated the changes in α-tubulin, β-actin, HSC70, and major capsid protein (MCP) during infection. β-Actin was selected for further study via co-immunoprecipitation analyses, which confirmed that the cellular β-actin interacts with the MCP protein of ISKNV in the infected zebrafish. These findings provide insight into the interactions between iridoviruses (especially ISKNV) and host, as well as the mechanism and pathogenesis of ISKNV infections.
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Affiliation(s)
- Xiao-Peng Xiong
- State Key Laboratory of Biocontrol/MOE Key Laboratory of Aquatic Product Safety, School of Life Sciences, Sun Yat-sen University, Guangzhou 510275, People's Republic of China
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Analysis of white spot syndrome virus envelope protein complexome by two-dimensional blue native/SDS PAGE combined with mass spectrometry. Arch Virol 2011; 156:1125-35. [DOI: 10.1007/s00705-011-0954-7] [Citation(s) in RCA: 17] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Academic Contribution Register] [Received: 10/19/2010] [Accepted: 02/18/2011] [Indexed: 01/09/2023]
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The role of aldehyde dehydrogenase and hsp70 in suppression of white spot syndrome virus replication at high temperature. J Virol 2011; 85:3517-25. [PMID: 21228234 DOI: 10.1128/jvi.01973-10] [Citation(s) in RCA: 47] [Impact Index Per Article: 3.4] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Academic Contribution Register] [Indexed: 01/31/2023] Open
Abstract
High temperature (32 to 33°C) has been shown to reduce mortality in white spot syndrome virus (WSSV)-infected shrimps, but the mechanism still remains unclear. Here we show that in WSSV-infected shrimps cultured at 32°C, transcriptional levels of representative immediate-early, early, and late genes were initially higher than those at 25°C. However, neither the IE1 nor VP28 protein was detected at 32°C, suggesting that high temperature might inhibit WSSV protein synthesis. Two-dimensional gel electrophoresis analysis revealed two proteins, NAD-dependent aldehyde dehydrogenase (ALDH) and the proteasome alpha 4 subunit (proteasome α4), that were markedly upregulated in WSSV-infected shrimps at 32°C. Reverse transcription-PCR (RT-PCR) analysis of members of the heat shock protein family also showed that hsp70 was upregulated at 32°C. When aldh, proteasome α4, and hsp70 were knocked down by double-stranded RNA interference and shrimps were challenged with WSSV, the aldh and hsp70 knockdown shrimps became severely infected at 32°C, while the proteasome α4 knockdown shrimps remained uninfected. Our results therefore suggest that ALDH and Hsp70 both play an important role in the inhibition of WSSV replication at high temperature.
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Liang Y, Cheng JJ, Yang B, Huang J. The role of F1 ATP synthase beta subunit in WSSV infection in the shrimp, Litopenaeus vannamei. Virol J 2010; 7:144. [PMID: 20591132 PMCID: PMC2906456 DOI: 10.1186/1743-422x-7-144] [Citation(s) in RCA: 33] [Impact Index Per Article: 2.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Academic Contribution Register] [Received: 04/22/2010] [Accepted: 06/30/2010] [Indexed: 11/23/2022] Open
Abstract
Background Knowledge of the virus-host cell interaction could inform us of the molecular pathways exploited by the virus. Studies on viral attachment proteins (VAPs) and candidate receptor proteins involved in WSSV infection, allow a better understanding of how these proteins interact in the viral life cycle. In this study, our aim was to find some host cellular membrane proteins that could bind with white spot syndrome virus (WSSV). Results Two proteins were evident by using a virus overlay protein binding assay (VOPBA) with WSSV. A protein with molecular weight 53 kDa, named BP53, was analyzed in this study, which was homologous with the F1-ATP synthase beta subunit by mass spectrometry analysis. Rapid amplification of cDNA ends (RACE) PCR was performed to identify the full-length cDNA of the bp53 gene. The resulting full-length gene consisted of 1836 bp, encoding 525 amino acids with a calculated molecular mass of 55.98 kDa. The deduced amino acid sequence contained three conserved domains of the F1-ATP synthase beta subunit. BP53 was therefore designated the F1-ATP synthase beta subunit of L. vannamei. The binding of WSSV to BP53 were also confirmed by competitive ELISA binding assay and co-immunoprecipitation on magnetic beads. To investigate the function of BP53 in WSSV infection, it was mixed with WSSV before the mixture was injected intramuscularly into shrimp. The resulting mortality curves showed that recombinant (r) BP53 could attenuate WSSV infection. Conclusions The results revealed that BP53 is involved in WSSV infection. Here is the first time showed the role of shrimp F1-ATP synthase beta subunit in WSSV infection.
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Affiliation(s)
- Yan Liang
- Key Laboratory of Sustainable Utilization of Marine Fisheries Resources, the Ministry of Agriculture; Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China
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Syed Musthaq S, Madhan S, Sahul Hameed A, Kwang J. Localization of VP28 on the baculovirus envelope and its immunogenicity against white spot syndrome virus in Penaeus monodon. Virology 2009; 391:315-24. [DOI: 10.1016/j.virol.2009.06.017] [Citation(s) in RCA: 58] [Impact Index Per Article: 3.6] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Academic Contribution Register] [Received: 05/05/2009] [Revised: 05/24/2009] [Accepted: 06/03/2009] [Indexed: 11/30/2022]
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