A novel cell line established from caudal fin tissue of Carassius auratus gibelio is susceptible to cyprinid herpesvirus 2 infection with the induction of apoptosis

Jianfei Lu; Dan Xu; Liqun Lu

doi:10.1016/j.virusres.2018.09.010

A novel cell line established from caudal fin tissue of Carassius auratus gibelio is susceptible to cyprinid herpesvirus 2 infection with the induction of apoptosis

Virus Res. 2018 Oct 15:258:19-27. doi: 10.1016/j.virusres.2018.09.010. Epub 2018 Sep 27.

Authors

Jianfei Lu¹, Dan Xu², Liqun Lu³

Affiliations

¹ National Pathogen Collection Center for Aquatic Animals, Shanghai Ocean University, Shanghai, PR China; Key Laboratory of Agriculture Ministry for Freshwater Aquatic Genetic Resources, Shanghai Ocean University, Shanghai, PR China; National Experimental Teaching Demonstration Center for Fishery Sciences, Shanghai Ocean University, Shanghai, PR China; Laboratory of Biochemistry and Molecular Biology, Department of Marine and Life Sciences, Ningbo University, Ningbo 315211, PR China.
² National Pathogen Collection Center for Aquatic Animals, Shanghai Ocean University, Shanghai, PR China; Key Laboratory of Agriculture Ministry for Freshwater Aquatic Genetic Resources, Shanghai Ocean University, Shanghai, PR China; National Experimental Teaching Demonstration Center for Fishery Sciences, Shanghai Ocean University, Shanghai, PR China.
³ National Pathogen Collection Center for Aquatic Animals, Shanghai Ocean University, Shanghai, PR China; Key Laboratory of Agriculture Ministry for Freshwater Aquatic Genetic Resources, Shanghai Ocean University, Shanghai, PR China; National Experimental Teaching Demonstration Center for Fishery Sciences, Shanghai Ocean University, Shanghai, PR China. Electronic address: lqlv@shou.edu.cn.

PMID: 30268797
DOI: 10.1016/j.virusres.2018.09.010

Abstract

Herpesviral haematopoietic necrosis induced by cyprinid herpesvirus 2 (CyHV-2) in Carassius auratus gibelio has caused huge economic losses in aquaculture. To better characterize this pathogen at molecular and cellular levels, it is necessary to study its pathogenesis in a sustainable cell line. In this study, a new cell line (GiCF) from the caudal ﬁn of Carassius auratus gibelio (C. a. gibelio) was established and characterised. Typical cytopathic effect represented by characteristic morphological changes was induced in the cells at the 7th day post infection with CyHV-2, which was isolated from diseased C. a. gibeli. The susceptibility of GiCF to CyHV-2 was further demonstrated by the presence of viral assembling factory in the cytoplasm unveiled by transmission electron microscopy, and viral protein synthesis through immunoﬂuorescence and Western blot assays. Sustainable propagation of CyHV-2 in GiCF had been demonstrated by virus infection and titration assays in a consecutive manner. The virus titer reached 10^4.9±0.22 TCID₅₀/ml, and infectious CyHV-2 was produced from the GiCF cells over 30 subcultures from the established cell lines. In addition, apoptosis was observed in the GiCF cells upon CyHV-2 infection by DAPI staining, TUNEL and Annexin V staining, and quantitative expression patterns of caspase genes suggested that caspase activation might involve in CyHV-2 induced apoptosis. In summary, a new cell line, GiCF, susceptible to CyHV-2 infection was established from caudal fin tissue of C. a. gibelio, and it should serve as an ideal infection platform to characterize CyHV-2 and its pathogenic effect including apoptosis.

Keywords: Apoptosis; Carassius auratus gibelio; Caudal ﬁn cell line; Cyprinid herpesvirus 2.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animal Fins / cytology*
Animals
Apoptosis*
Caspases / metabolism
Cell Culture Techniques / methods*
Cell Line*
Fish Diseases / virology
Goldfish*
Herpesviridae / pathogenicity*
Herpesviridae Infections / veterinary
Herpesviridae Infections / virology
Necrosis

Substances

Caspases

Supplementary concepts

Cyprinid herpesvirus 2