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应用 OpenArray®平台分析感染神经坏死病毒的塞内加尔鳎中的免疫基因表达。

Immunogene expression analysis in betanodavirus infected-Senegalese sole using an OpenArray® platform.

机构信息

Universidad de Málaga, Instituto de Biotecnología y Desarrollo Azul, IBYDA, Departamento de Microbiología, Facultad de Ciencias, Málaga, Spain.

Universidade de Santiago de Compostela, Instituto de Acuicultura, Departamento de Microbiología y Parasitología, Santiago de Compostela, Spain.

出版信息

Gene. 2021 Mar 30;774:145430. doi: 10.1016/j.gene.2021.145430. Epub 2021 Jan 11.

Abstract

The transcriptomic response of Senegalese sole (Solea senegalensis) triggered by two betanodaviruses with different virulence to that fish species has been assessed using an OpenArray® platform based on TaqMan™ quantitative PCR. The transcription of 112 genes per sample has been evaluated at two sampling times in two organs (head kidney and eye/brain-pooled samples). Those genes were involved in several roles or pathways, such as viral recognition, regulation of type I (IFN-1)-dependent immune responses, JAK-STAT cascade, interferon stimulated genes, protein ubiquitination, virus responsive genes, complement system, inflammatory response, other immune system effectors, regulation of T-cell proliferation, and proteolysis and apoptosis. The highly virulent isolate, wSs160.3, a wild type reassortant containing a RGNNV-type RNA1 and a SJNNV-type RNA2 segments, induced the expression of a higher number of genes in both tested organs than the moderately virulent strain, a recombinant harbouring mutations in the protruding domain of the capsid protein. The number of differentially expressed genes was higher 2 days after the infection with the wild type isolate than at 3 days post-inoculation. The wild type isolate also elicited an exacerbated interferon 1 response, which, instead of protecting sole against the infection, increases the disease severity by the induction of apoptosis and inflammation-derived immunopathology, although inflammation seems to be modulated by the complement system. Furthermore, results derived from this study suggest a potential important role for some genes with high expression after infection with the highly virulent virus, such as rtp3, sacs and isg15. On the other hand, the infection with the mutant does not induce immune response, probably due to an altered recognition by the host, which is supported by a different viral recognition pathway, involving myd88 and tbkbp1.

摘要

采用基于 TaqMan 定量 PCR 的 OpenArray®平台,评估了两种对塞内加尔比目鱼(Solea senegalensis)毒力不同的 betanodaviruses 对该鱼类物种引发的转录组反应。在两个器官(头肾和眼/脑合并样本)的两个采样时间评估了每个样本的 112 个基因的转录。这些基因参与了几种作用或途径,如病毒识别、I 型(IFN-1)依赖性免疫反应的调节、JAK-STAT 级联、干扰素刺激基因、蛋白质泛素化、病毒反应基因、补体系统、炎症反应、其他免疫系统效应物、T 细胞增殖的调节以及蛋白水解和细胞凋亡。高毒力分离株 wSs160.3 是一种含有 RGNNV 型 RNA1 和 SJNNV 型 RNA2 节段的野生型重组体,比含有衣壳蛋白突出结构域突变的重组体在两个测试器官中诱导了更多基因的表达。感染野生型分离株后 2 天的差异表达基因数量高于接种后 3 天。野生型分离株还引起了干扰素 1 反应的加剧,这不仅没有保护比目鱼免受感染,反而通过诱导细胞凋亡和炎症引起的免疫病理学增加疾病的严重程度,尽管炎症似乎受到补体系统的调节。此外,这项研究的结果表明,一些在感染高毒力病毒后高表达的基因可能具有重要作用,例如 rtp3、sacs 和 isg15。另一方面,感染突变体不会引起免疫反应,可能是由于宿主的识别发生改变,这得到了涉及 myd88 和 tbkbp1 的不同病毒识别途径的支持。

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