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冠状病毒:感染类似 IS-1494 变异 2 型(IS-1494)的禽传染性支气管炎病毒的鸡肾组织的蛋白质组学分析。

Coronavirus: proteomics analysis of chicken kidney tissue infected with variant 2 (IS-1494)-like avian infectious bronchitis virus.

机构信息

Proteomics Research Center, Faculty of Paramedical Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

National Reference Laboratory, Diagnosis & Applied Studies Center, Iran Veterinary Organization, Tehran, Iran.

出版信息

Arch Virol. 2021 Jan;166(1):101-113. doi: 10.1007/s00705-020-04845-7. Epub 2020 Oct 20.

Abstract

Avian infectious bronchitis virus is one of the most important gammacoronaviruses, which causes a highly contagious disease. In this study, we investigated changes in the proteome of kidney tissue of specific-pathogen-free (SPF) chickens that were infected with an isolate of the nephrotropic variant 2 genotype (IS/1494/06) of avian coronavirus. Twenty 1-day-old SPF White Leghorn chickens were randomly divided into two groups, each comprising 10 chickens, which were kept in separate positive-pressure isolators. Chickens in group A served as a virus-free control group up to the end of the experiment, whereas chickens in group B were inoculated with 0.1 ml of 10 EID of the IBV/chicken/Iran/UTIVO-C/2014 isolate of IBV, and kidney tissue samples were collected at 2 and 7 days post-inoculation (dpi) from both groups. Sequencing of five protein spots at 2 dpi and 22 spots at 7 dpi that showed differential expression by two-dimensional electrophoresis (2DE) along with fold change greater than 2 was done by MS-MALDI/TOF/TOF. Furthermore, the corresponding protein-protein interaction (PPI) networks at 2 and 7 dpi were identified to develop a detailed understanding of the mechanism of molecular pathogenesis. Topological graph analysis of this undirected PPI network revealed the effect of 10 genes in the 2 dpi PPI network and nine genes in the 7 dpi PPI network during virus pathogenesis. Proteins that were found by 2DE analysis and MS/TOF-TOF mass spectrometry to be down- or upregulated were subjected to PPI network analysis to identify interactions with other cellular components. The results show that cellular metabolism was altered due to viral infection. Additionally, multifunctional heat shock proteins with a significant role in host cell survival may be employed circuitously by the virus to reach its target. The data from this study suggest that the process of pathogenesis that occurs during avian coronavirus infection involves the regulation of vital cellular processes and the gradual disruption of critical cellular functions.

摘要

禽传染性支气管炎病毒是最重要的γ冠状病毒之一,可引起高度传染性疾病。在这项研究中,我们研究了感染禽冠状病毒肾病变型 2 基因型(IS/1494/06)分离株的无特定病原体(SPF)鸡肾脏组织蛋白质组的变化。将 20 只 1 日龄 SPF 白来航鸡随机分为两组,每组 10 只,分别饲养在独立的正压隔离器中。A 组鸡作为病毒对照组,直至实验结束,B 组鸡接种 0.1ml 10EID 的 IBV/chicken/Iran/UTIVO-C/2014 株 IBV,分别于接种后 2 天和 7 天从两组收集肾脏组织样本。通过二维电泳(2DE)结合差异表达倍数大于 2 对 2 dpi 时的 5 个蛋白质斑点和 7 dpi 时的 22 个斑点进行测序,采用 MS-MALDI/TOF/TOF。此外,还鉴定了 2 dpi 和 7 dpi 时相应的蛋白质-蛋白质相互作用(PPI)网络,以深入了解分子发病机制。对无向 PPI 网络的拓扑图分析揭示了病毒发病过程中 2 dpi PPI 网络中的 10 个基因和 7 dpi PPI 网络中的 9 个基因的影响。通过 2DE 分析和 MS/TOF-TOF 质谱鉴定出下调或上调的蛋白质,并用 PPI 网络分析鉴定与其他细胞成分的相互作用。结果表明,病毒感染导致细胞代谢发生改变。此外,多功能热休克蛋白在宿主细胞存活中具有重要作用,可能被病毒间接利用以达到其靶标。本研究数据表明,禽冠状病毒感染过程中的发病机制涉及关键细胞过程的调节和关键细胞功能的逐渐破坏。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c7f9/7574675/897b18913b32/705_2020_4845_Fig1_HTML.jpg

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