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Characterization of three novel cell lines derived from the brain of spotted sea bass: Focusing on cell markers and susceptibility toward iridoviruses

文献类型： 外文期刊

作者： Liu, Zhenxing 1 ; Ma, Yanping 1 ; Hao, Le 1 ;

作者机构： 1.Guangdong Acad Agr Sci, Inst Anim Hlth, Guangzhou 510640, Peoples R China

2.Key Lab Livestock Dis Prevent Guangdong Prov, Guangzhou 510640, Peoples R China

3.Minist Agr & Rural Affairs, Sci Observat & Expt Stn Vet Drugs & Diagnost Tech, Guangzhou 510640, Peoples R China

4.Collaborat Innovat Ctr GDAAS, Guangzhou, Peoples R China

关键词： Cell line; Cell marker; Iridovirus; Megalocytivirus; Ranavirus; Lateolabrax maculatus; Type I IFN pathway

期刊名称：FISH & SHELLFISH IMMUNOLOGY （ 影响因子：4.622； 五年影响因子：4.799 ）

ISSN： 1050-4648

年卷期： 2022 年 130 卷

页码：

收录情况： SCI

摘要： Despite tens of cell lines originating from fish brain tissue have been constructed, little is known about the definite cell types they belong to. Whether fish cell lines derived from the brain shares similar characteristics is not well-answered yet. Here, we constructed three cell lines designated as LMB-S, LMB-M, LMB-L using brain tissue of spotted sea bass (Lateolabrax maculatus). Among them, LMB-L was identified as astroglia-like cells considering the high expression of GFAP, DCX, PTX, S100b, which are regarded as astrocyte-specific or astrocyte-associated cell markers. LMB-M exhibited smooth muscle-like features showing strong expression of LMOD1, SLAMP, M-cadherin, MGP, which are confirmed as muscle-restricted or myogenesis-involved cell markers. Although LMB-S was not definitely identified, it appeared an activation of WNT/beta-catenin pathway. Besides the distinct expression profiles of cell markers, the three cell lines also presented differences in transfection efficiency and susceptibility to iridovirus infection. Relying on the established cell lines, a novel megalocytivirus, named LMIV (Lateolabrax maculatus iridovirus), was first isolated from diseased spotted sea bass. Genetic analysis of major capsid protein (MCP) and adenosine triphosphatase (ATPase) manifested that LMIV was clearly distin-guishable from other representative teleost iridoviruses. Further investigations revealed that LMIV could repli-cate most efficiently in LMB-L cells obtaining the highest viral load (2.16 x 1010 copy/mL). By contrast, LMB-S cells gave rise to the highest viral load up to 3.86 x 108 copy/mL, when the three cell lines were infected with MRV, a newly emerged ranavirus. Moreover, LMIV infection caused lots of cells to be detached from monolayers, generating adherent and non-adherent cells. An opposite expression profiling of type I IFN pathway-related genes (JAK1, STAT1, STAT2, IRF9, Mx1) was found between adherent and non-adherent cells. Combined with the analysis of MCP gene expression, it is speculated that inhibiting type I IFN pathway in non-adherent cells allowed the facilitation of virus duplication. Taken together, the present study broadens our understanding about the diversity of cell lines derived from fish brain tissue and screening cells more susceptible to virus is not only meaningful for the development of vaccine, but also provide clues for further clarification of cell-iridovirus interactions.

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