Innate response of rainbow trout gill epithelial (RTgill-W1) cell line to ultraviolet-inactivated VHSV and FliC and rhabdovirus infection

Ehab Misk , Paul Huber , Janet I. MacInnes , Sherif M. Sherif , Mohammed Abo-Ismail , John S. Lumsden
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引用次数: 2

Abstract

Gills reportedly play a crucial role in induction of an antiviral immune response in fish. We investigated the expression of innate response genes in the rainbow trout gill epithelial cell line RTgill-W1 36 h after pretreatment with ultraviolet-inactivated viral hemorrhagic septicemia virus (UV-VHSV), flagellin C protein from Edwardsiella tarda (FliC), VHSV and SVCV using an Agilent 4 × 44k cGRASP salmonid microarray. RTgill-W1 cells pretreated with UV-VHSV, triggered an independent gene expression profile from those treated with a recombinant flagellin C protein from Edwardsiella tarda. In addition, exposure of RTgill-W1 cells to live viruses spring viremia of carp virus and viral hemorrhagic septicemia virus induced a less robust transcriptional change of 24 and 22 gene probes, respectively, when compared to 123 genes for UV-VHSV. Further the pretreatment of RTgill-W1 cells with (UV-VHSV) significantly reduced VHSV genome copy number at 6 d post infection (dpi) relative to the FliC-treated and untreated control. A quantitative PCR was used to study the transcriptional modulation of a set of 25 innate immune-related genes highlighted by the microarray data and a panel of 7 established antiviral genes in the protected cells. Notably, the expression of ifn1, ifn2, mx1 and mx3 were expressed more in untreated cells than in UV-VHSV-treated cells where virus replication was inhibited. The results from this study shed light on the mechanisms and pathways used by teleost gill epithelium innate immunity in combating viral and bacterial infection.

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虹鳟鱼鳃上皮细胞(RTgill-W1)对紫外线灭活VHSV、flc和横纹肌病毒感染的先天应答
据报道,鳃在诱导鱼类抗病毒免疫反应中起着至关重要的作用。采用安捷伦4 × 44k cGRASP鲑鱼微阵列技术,研究了经紫外线灭活的病毒性出血性败血症病毒(UV-VHSV)、迟爱德华菌(FliC)、VHSV和SVCV的鞭毛蛋白C蛋白预处理36小时后虹鳟鱼鳃上皮细胞rtggil - w1的先天反应基因表达。经过UV-VHSV预处理的rtggil - w1细胞,与经过迟缓爱德华菌重组鞭毛蛋白C处理的细胞相比,触发了独立的基因表达谱。此外,RTgill-W1细胞暴露于活病毒(鲤鱼春季病毒血症和病毒性出血性败血症病毒)中,分别诱导了24个和22个基因探针的转录变化,而与UV-VHSV的123个基因相比,它们的转录变化较弱。此外,RTgill-W1细胞与(UV-VHSV)预处理在感染后6天显著减少VHSV基因组拷贝数(dpi)相对于flicv处理和未处理的对照。利用定量PCR技术研究了微阵列数据显示的25个先天免疫相关基因和7个已建立的抗病毒基因在受保护细胞中的转录调节。值得注意的是,在未处理的细胞中,ifn1、ifn2、mx1和mx3的表达比在抑制病毒复制的uv - vhsv处理的细胞中表达得更多。本研究的结果揭示了硬骨鱼鳃上皮先天免疫对抗病毒和细菌感染的机制和途径。
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