| 鲤疱疹病毒Ⅱ型对异育银鲫背鳍细胞的显微形态与免疫基因表达水平的影响 |
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| 引用本文： | 夏思瑶,王浩,Patorida Podok,许丹,姜有声,吕利群,佐野元彦.鲤疱疹病毒Ⅱ型对异育银鲫背鳍细胞的显微形态与免疫基因表达水平的影响[J].水产学报,2016,40(12):1915-1922. |
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| 作者姓名： | 夏思瑶 王浩 Patorida Podok 许丹 姜有声 吕利群 佐野元彦 |
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| 作者单位： | 1. 上海海洋大学水产与生命学院,国家水生动物病原库,上海201306;2. 日本国家水产研究所渔业研究中心,东京516-0193 |
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| 基金项目： | 现代农业产业技术体系建设专项资金(CARS-46-12) |
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| 摘 要： | 为研究鲤疱疹病毒Ⅱ型(Cy HV-2)体外感染复制特征以及异育银鲫抗病毒免疫应答反应。本实验采用组织块培养法建立了异育银鲫背鳍细胞的原代培养体系。结果显示,在10 d左右可观察到组织块迁移分离出新的单层细胞,3周左右细胞可覆盖底部面积为25cm2培养瓶的底部;经Cy HV-2悬液感染离体培养的原代细胞,3 d后病毒滴度增殖至106拷贝/m L;在病毒感染6 d后出现典型的细胞病变效应;Cy HV-2感染原代细胞后,分析前期通过鱼体水平实验鉴定出的与该病毒感染相关的免疫基因:PNP5a、MPO、MHCⅠ、LYZ-C、IL-11、ITLN、PNP5a和DUSP,Real-time Rt-PCR结果显示大部分基因在细胞水平均有显著性的上调,与鱼体水平实验结果一致。本研究建立了原代培养的异育银鲫背鳍细胞,用于构建体外感染Cy HV-2病毒的细胞模型,为深入研究Cy HV-2的感染复制规律及其与宿主的相互作用关系,以及细胞水平筛选抗病毒药物实验奠定了基础。
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| 关 键 词： | 异育银鲫 鲤疱疹病毒Ⅱ型 背鳍细胞 原代培养 Real time Rt-PCR |
| 收稿时间： | 2016/1/19 0:00:00 |
| 修稿时间： | 2016/5/30 0:00:00 |
The expression analysis of immune genes and microscopic morphology of CCF cells in response to Cyprinid herpesvirus 2 infection |
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| XIA Siyao,WANG Hao,Patorida Podok,XU Dan,JIANG Yousheng,L&#; Liqun and Motohiko Sano.The expression analysis of immune genes and microscopic morphology of CCF cells in response to Cyprinid herpesvirus 2 infection[J].Journal of Fisheries of China,2016,40(12):1915-1922. |
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| Authors: | XIA Siyao WANG Hao Patorida Podok XU Dan JIANG Yousheng L&#; Liqun and Motohiko Sano |
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| Institution: | National Pathogen Collection Center of Aquatic Animals, College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China,National Pathogen Collection Center of Aquatic Animals, College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China,National Pathogen Collection Center of Aquatic Animals, College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China,National Pathogen Collection Center of Aquatic Animals, College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China,National Pathogen Collection Center of Aquatic Animals, College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China,National Pathogen Collection Center of Aquatic Animals, College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China and Fisheries Research Agency, National Research Institute of Aquaculture, Tokyo 516-0193, Japan |
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| Abstract: | The aims of the current study were to investigated the primary culture of dorsal fin from Prussian carp, Carassius auratus gibelio and its susceptibility to Cyprinid herpesvirus 2 due to the lack of commercial cell line supporting CyHV-2 infection. The cells detached from the tissues in 10 days and fully covered the bottom of 25 cm2 culture bottle in 3 weeks. Cyprinid herpesvirus 2 (CyHV-2) continuously propagated in the cultured cells, as confirmed by a quantitive real time PCR, with the highest viral concentration of 106 copies/mL at 3 days post infection. In addition, cell death was observed in the dorsal fin cells upon CyHV-2 infection. Cytopathic effects (CPE) were observed as early as 6 days post-inoculation, and as the infection progressed, CPE became more apparent with cell debris and cellular exudates in inoculated cultures. The expression levels of 8 immune important genes including L-11, ITLN, PNP5a, MPO, KRT-8, DUSP-1, MHC I and LYZ C were significant different after viral challenge, which were determined to be involved in host response to CyHV-2 infection previously. The results above should be helpful in further studies on CyHV-2, including elucidation of pathogenesis, host-pathogen interaction and development of anti-viral drugs that ultimately lead to prevention of this viral disease. |
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| Keywords: | Carassius auratus gibelio CyHV-2 dorsal fin cells primary culture Real time Rt-PCR |
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