Establishment and characterization of a Gobiocypris rarus cell line for studying type II grass carp reovirus infection

IF 3.7 2区农林科学 Q1 FISHERIES

Aquaculture Reports Pub Date : 2025-08-29 DOI:10.1016/j.aqrep.2025.103065

Wenji Huang , Sijia Wan , Ling Shao

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引用次数: 0

Abstract

The type II Grass Carp Reovirus (GCRV-II) causes severe hemorrhagic disease with high mortality in economically important fish species. However, existing cell culture systems exhibits neither efficient viral replication nor observable cytopathic effects. In this study, a novel cell line (GRLJ) was established utilizing cells obtained from the lower jaw of Gobiocypris rarus (rare minnow), which was characterized in terms of morphology, proliferation, chromosomes, transfection efficiency, susceptibility to GCRV-II, and innate immune response. GRLJ cells with an epithelioid phenotype infected with GCRV-II exhibited a significant viral particle in the cytoplasm, and upregulation of viral genes (vp4, vp56). High-titer infection (1.9 × 10⁴ copies/μL) caused complete lethality in rare minnow by 10 days post-infection (dpi), with evident histopathology. The calculated LD₅₀ for GRLJ-derived GCRV-II was 2.6 × 10 ³ copies/μL. The expression of immune-related genes, such as tlr3, tbk1, ifnα, ifn2, p65 and pkr were significantly upregulated at 3 and 7 dpi. These results highlight the potential of GRLJ cells as a model for studying GCRV-II replication and host-pathogen interactions.

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用于研究II型草鱼呼肠孤病毒感染的稀有虾蛄细胞系的建立和鉴定

II型草鱼呼肠孤病毒（GCRV-II）在经济上重要的鱼类中引起严重的出血性疾病，死亡率高。然而，现有的细胞培养系统既没有有效的病毒复制，也没有可观察到的细胞病变效应。本研究利用稀有小鱼（Gobiocypris rarus, Gobiocypris rarus）下颚的细胞建立了一种新的细胞系（GRLJ），该细胞系在形态、增殖、染色体、转染效率、对GCRV-II的易感性和先天免疫反应等方面具有特征。具有上皮样表型的GRLJ细胞感染GCRV-II后，细胞质中出现了显著的病毒颗粒，病毒基因（vp4, vp56）上调。高滴度感染（1.9 × 10⁴拷贝/μL）导致罕见的小鱼在感染后10天（dpi）完全死亡，具有明显的组织病理学。grlj衍生的GCRV-II的计算LD₅0为2.6 × 10 ³ copies/μL。免疫相关基因tlr3、tbk1、ifnα、ifn2、p65和pkr的表达在3和7 dpi时显著上调。这些结果突出了GRLJ细胞作为研究GCRV-II复制和宿主-病原体相互作用模型的潜力。

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来源期刊

Aquaculture Reports Agricultural and Biological Sciences-Animal Science and Zoology

CiteScore

5.90

自引率

8.10%

发文量

469

审稿时长

77 days

期刊介绍： Aquaculture Reports will publish original research papers and reviews documenting outstanding science with a regional context and focus, answering the need for high quality information on novel species, systems and regions in emerging areas of aquaculture research and development, such as integrated multi-trophic aquaculture, urban aquaculture, ornamental, unfed aquaculture, offshore aquaculture and others. Papers having industry research as priority and encompassing product development research or current industry practice are encouraged.