鲤鱼水肿病毒(基因组 I)在鲤鱼原代细胞单层中生长的潜力,以及对细胞分析的进一步影响。

IF 2.2 3区 农林科学 Q2 FISHERIES Journal of fish diseases Pub Date : 2024-02-29 DOI:10.1111/jfd.13934
Ehdaa Eltayeb Eltigani Abdelsalam, Zuzana Bláhová, Ali Asghar Baloch, Veronika Piačková
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引用次数: 0

摘要

鲤鱼水肿病毒(CEV)具有不同的分子鉴定病毒变异基因组,分别称为 I、IIa 和 IIb。由于无法在体外繁殖 CEV,限制了了解其与宿主细胞相互作用的研究。在此,我们利用在捷克共和国自然疫情中收集到的属于基因组 I 的病毒分离株,在鲤鱼原代细胞、鲤鱼脑细胞(CCB)和鲤鱼上皮乳头状瘤细胞(EPC)的单层细胞中进行常规 CEV 培养。观察并记录了受影响细胞的细胞病理效应(CPE)。在 CEV 接种体的系列稀释下评估细胞存活率。分别通过 qPCR 和透射电子显微镜对病毒进入细胞进行量化和可视化。研究结果表明,原发性鳃上皮可能是最适合 CEV 体外生长的基质。头部肾脏和脾脏的细胞有利于病毒进入,显微镜下可确认CPE和细胞质多形病毒颗粒的存在。躯干肾脏和性腺的细胞不太可能允许病毒细胞进入并形成 CPE。虽然在细胞系中培养 CEV 尚无定论,但 EPC 细胞是允许 CEV 的。鲤鱼原代细胞的单层细胞显示了培养 CEV 的前景,可用于详细研究细胞结合和反应的基本机制。
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Potential to grow carp oedema virus (genogroup I) in monolayers of carp-derived primary cells with further implication in cell analysis

Carp oedema virus (CEV) has distinct molecularly identified genogroups of viral mutations, denoted as I, IIa, and IIb. Failure to propagate CEV in vitro limits studies towards understanding its interactions with host cells. Here, virus isolates belonging to genogroup I collected during natural outbreaks in the Czech Republic were employed for routine CEV cultivation in monolayers of carp-derived primary cells, common carp brain (CCB) cells, and epithelioma papulosum cyprinid (EPC) cells. Induction of cytopathic effects (CPEs) was observed and recorded in affected cells. Cell survival rate was evaluated under serial dilutions of the CEV inoculum. Virus cell entry was quantified and visualized by qPCR and transmission electron microscopy, respectively. Study findings indicate primary gills epithelia likely present the most suitable matrix for CEV growth in vitro. Cells of the head kidney and spleen facilitate virus entry with microscopically confirmed CPEs and the presence of cytoplasmic pleomorphic virus particles. Cells of the trunk kidney and gonads are unlikely to permit virus cell entry and CPEs development. Although CEV cultivation in cell lines was inconclusive, EPC cells were CEV permissible. Monolayers of carp-derived primary cells show promise for CEV cultivation that could enable elaborate study of mechanisms underlying cellular binding and responses.

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来源期刊
Journal of fish diseases
Journal of fish diseases 农林科学-海洋与淡水生物学
CiteScore
4.60
自引率
12.00%
发文量
170
审稿时长
6 months
期刊介绍: Journal of Fish Diseases enjoys an international reputation as the medium for the exchange of information on original research into all aspects of disease in both wild and cultured fish and shellfish. Areas of interest regularly covered by the journal include: -host-pathogen relationships- studies of fish pathogens- pathophysiology- diagnostic methods- therapy- epidemiology- descriptions of new diseases
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