Development and characterization of monoclonal antibodies to spring viraemia of carp virus
文献类型：期刊论文
| 作者 | Chen, Zhong-Yuan1; Liu, Hong2; Li, Zheng-Qiu1; Zhang, Qi-Ya1 |
| 刊名 | VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY
 |
| 出版日期 | 2008-06-15 |
| 卷号 | 123期号:3-4页码:266-276 |
| ISSN号 | 0165-2427 |
| 关键词 | monoclonal antibodies fish rhabdovirus spring viraemia of carp virus (SVCV0504) flow cytometry |
| 通讯作者 | Zhang, QY, Wuhan Univ, State Lab Freshwater Ecol & Biotechnol, Inst Hydrobiol, Grad Sch, Wuhan 430072, Peoples R China |
| 中文摘要 | Five monoclonal antibodies (mAbs) against spring viraemia of carp (SVCV0504, isolated from common carp in China) were produced from mice immunized with purified virus preparations. The virion of SVCV contains five structural proteins, representing the nucleoprotein (N), phosphoprotein (P), matrix protein (M), glycoprotein (G) and RNA-dependent RNA polymerase (Q. Western blotting analysis revealed that three mAbs (1145, IE10, and 11-17) recognized specifically to a single protein of 47 kDa (N), the mAb 3G4 reacted with, two SVCV0504 proteins of 69 kDa (G) and 47 kDa (N), while the mAb 1A9 reacted with three SVCV0504 proteins of 69 kDa (G), 50 kDa (P), and 47 kDa (N). By indirect ELISA, two mAbs (1H5 and 11-17) showed cross-reactivity with pike fry rhabdovirus (PFRV), but no cross-reactions with the Siniperca chuatsi rhabdovirus (SCRV), Scophthalmus maximus rhabdovirus (SMRV), Paralichthys olivaceus rhabdovirus (PoRV) were demonstrated with the five mAbs. Indirect immunofluorescence showed intense fluorescence in the cytoplasm of the SVCV0504-infected epithelioma papulosum cyprini (EPC) cells in areas corresponding to the location of granular structures. The sucrose gradient-purified SVCV0504 particles could be detected successfully by these mAbs using immunodot blotting. mAb 1A9 could completely neutralize 100 TCID50 (50% tissue culture infective dose) of SVCV0504 at a dilution of 1:8. This is the first report of development of the neutralizing mAbs against SVCV. The mAb 1A9 was analyzed further and could be used to successfully detect viral antigens in the infected-EPC cell cultures or in cryosections from experimentally infected crucian carp (Carassius auratus) by immunohistochemistry assay. Furthermore, a flow cytometry procedure for the detection and quantification of cytoplasmic SVCV0504 in cell cultures was developed with mAb 1A9. At 28 h after inoculation with the virus (0.01 PFU/cell), 10.12% of infected cells could be distinguished from the uninfected cells. These mAbs will be useful in diagnostic test development and pathogenesis studies for fish rhabdovirus. (c) 2008 Elsevier B.V. All rights reserved. |
| 英文摘要 | Five monoclonal antibodies (mAbs) against spring viraemia of carp (SVCV0504, isolated from common carp in China) were produced from mice immunized with purified virus preparations. The virion of SVCV contains five structural proteins, representing the nucleoprotein (N), phosphoprotein (P), matrix protein (M), glycoprotein (G) and RNA-dependent RNA polymerase (Q. Western blotting analysis revealed that three mAbs (1145, IE10, and 11-17) recognized specifically to a single protein of 47 kDa (N), the mAb 3G4 reacted with, two SVCV0504 proteins of 69 kDa (G) and 47 kDa (N), while the mAb 1A9 reacted with three SVCV0504 proteins of 69 kDa (G), 50 kDa (P), and 47 kDa (N). By indirect ELISA, two mAbs (1H5 and 11-17) showed cross-reactivity with pike fry rhabdovirus (PFRV), but no cross-reactions with the Siniperca chuatsi rhabdovirus (SCRV), Scophthalmus maximus rhabdovirus (SMRV), Paralichthys olivaceus rhabdovirus (PoRV) were demonstrated with the five mAbs. Indirect immunofluorescence showed intense fluorescence in the cytoplasm of the SVCV0504-infected epithelioma papulosum cyprini (EPC) cells in areas corresponding to the location of granular structures. The sucrose gradient-purified SVCV0504 particles could be detected successfully by these mAbs using immunodot blotting. mAb 1A9 could completely neutralize 100 TCID50 (50% tissue culture infective dose) of SVCV0504 at a dilution of 1:8. This is the first report of development of the neutralizing mAbs against SVCV. The mAb 1A9 was analyzed further and could be used to successfully detect viral antigens in the infected-EPC cell cultures or in cryosections from experimentally infected crucian carp (Carassius auratus) by immunohistochemistry assay. Furthermore, a flow cytometry procedure for the detection and quantification of cytoplasmic SVCV0504 in cell cultures was developed with mAb 1A9. At 28 h after inoculation with the virus (0.01 PFU/cell), 10.12% of infected cells could be distinguished from the uninfected cells. These mAbs will be useful in diagnostic test development and pathogenesis studies for fish rhabdovirus. (c) 2008 Elsevier B.V. All rights reserved. |
| 学科主题 | Immunology; Veterinary Sciences |
| WOS标题词 | Science & Technology ; Life Sciences & Biomedicine |
| 类目[WOS] | Immunology ; Veterinary Sciences |
| 研究领域[WOS] | Immunology ; Veterinary Sciences |
| 关键词[WOS] | HEMATOPOIETIC NECROSIS VIRUS ; SCOPHTHALMUS-MAXIMUS RHABDOVIRUS ; FLOUNDER PARALICHTHYS-OLIVACEUS ; CYPRINUS-CARPIO ; COMMON CARP ; FLOW-CYTOMETRY ; GLYCOPROTEIN GENE ; NORTH-AMERICA ; 1ST REPORT ; FISH |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000256747800011 |
| 公开日期 | 2010-10-13 |
| 源URL | [http://ir.ihb.ac.cn/handle/152342/8090]  |
| 专题 | 水生生物研究所_中科院水生所知识产出（2009年前）_期刊论文 |
| 作者单位 | 1.Wuhan Univ, State Lab Freshwater Ecol & Biotechnol, Inst Hydrobiol, Grad Sch, Wuhan 430072, Peoples R China 2.Shenzhen Exit & Entry Inspect & Quarantine Bur, Key Lab Aquat Anim Dis, Shenzhen 518001, Peoples R China |
| 推荐引用方式 GB/T 7714 | Chen, Zhong-Yuan,Liu, Hong,Li, Zheng-Qiu,et al. Development and characterization of monoclonal antibodies to spring viraemia of carp virus[J]. VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY,2008,123(3-4):266-276. |
| APA | Chen, Zhong-Yuan,Liu, Hong,Li, Zheng-Qiu,&Zhang, Qi-Ya.(2008).Development and characterization of monoclonal antibodies to spring viraemia of carp virus.VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY,123(3-4),266-276. |
| MLA | Chen, Zhong-Yuan,et al."Development and characterization of monoclonal antibodies to spring viraemia of carp virus".VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY 123.3-4(2008):266-276. |
入库方式： OAI收割
来源：水生生物研究所
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