Elucidation of the Host Bronchial Lymph Node miRNA Transcriptome Response to Bovine Respiratory Syncytial Virus.
Elucidation of the Host Bronchial Lymph Node miRNA Transcriptome Response to Bovine Respiratory Syncytial Virus.
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DOI：
10.3389/fgene.2021.633125
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发表时间：
2021
影响因子：
3.7
通讯作者：
Waters SM
中科院分区：
文献类型：
--
作者：
Johnston D;Earley B;McCabe MS;Kim J;Taylor JF;Lemon K;McMenamy M;Duffy C;Cosby SL;Waters SM
牛呼吸道疾病(BRD)导致大量的发病率和死亡率，影响到所有年龄段的牛。BRD的主要原因之一是对牛呼吸道合胞病毒(BRSV)的初始炎症反应。MicroRNAs是一种新出现的非编码小RNA，调节多种生物学过程，与多种炎症性疾病密切相关。本研究的目的是阐明牛支气管淋巴结miRNA转录组在实验性病毒攻击后对BRSV的响应变化。将12头荷斯坦-弗里西亚犊牛随机分为两组，每组12头，分别接种牛肾综合征出血热病毒(BRSV)103.5 TCID50/ml×15ml(n=12)，或模拟接种无菌磷酸盐缓冲盐水(n=6)。每天进行临床体征评分，并在激发后第7天处死小牛。收集支气管淋巴结，进行RNA提取和测序(75bp)。使用miRDeep2程序包，使用UMD3.1参考基因组和来自miRBase数据库(版本22)的牛成熟miRNA序列，生成已知miRNAs的读取计数。Edger软件进行差异表达分析，TargetScan软件分析差异表达的miRNAs的靶基因。目的基因的丰富途径和基因本体论的研究使用独创性路径分析(QIGEN)。基于miRNA基因表达变化的多维标度(MDS)显示，尽管疾病的临床表现很轻微，但BRSV感染牛犊和对照牛犊之间存在明显的分离。在攻毒组和对照组之间共检测到119个DE miRNAs(P<；0.0 5，FDR<；0.1，倍数变化>；1.5)。DE miRNAs被预测针对465个基因，这些基因以前在BRSV攻击的小牛和对照小牛之间的支气管淋巴组织中被发现是DE。在DE预测的目标基因中，455个具有与相应DE miRNAs相反的折叠变化。在DE预测的靶基因中，有8条途径与其对应的DE miRNA相反折叠变化，包括：粒细胞和无粒细胞的黏附和滞育，干扰素信号转导和病原体识别受体在识别细菌和病毒中的作用。预计增加的功能包括：T细胞反应、白细胞凋亡、细胞免疫反应和刺激细胞。病原体识别和细胞毒性T细胞的增殖对于病毒的识别和随后的消除至关重要。
Bovine respiratory disease (BRD) causes substantial morbidity and mortality, affecting cattle of all ages. One of the main causes of BRD is an initial inflammatory response to bovine respiratory syncytial virus (BRSV). MicroRNAs are novel and emerging non-coding small RNAs that regulate many biological processes and are implicated in various inflammatory diseases. The objective of the present study was to elucidate the changes in the bovine bronchial lymph node miRNA transcriptome in response to BRSV following an experimental viral challenge. Holstein-Friesian calves were either administered a challenge dose of BRSV (103.5 TCID50/ml × 15 ml) (n = 12) or were mock inoculated with sterile phosphate buffered saline (n = 6). Daily scoring of clinical signs was performed and calves were euthanized at day 7 post-challenge. Bronchial lymph nodes were collected for subsequent RNA extraction and sequencing (75 bp). Read counts for known miRNAs were generated using the miRDeep2 package using the UMD3.1 reference genome and the bovine mature miRNA sequences from the miRBase database (release 22). EdgeR was used for differential expression analysis and Targetscan was used to identify target genes for the differentially expressed (DE) miRNAs. Target genes were examined for enriched pathways and gene ontologies using Ingenuity Pathway Analysis (Qiagen). Multi-dimensional scaling (MDS) based on miRNA gene expression changes, revealed a clearly defined separation between the BRSV challenged and control calves, although the clinical manifestation of disease was only mild. One hundred and nineteen DE miRNAs (P < 0.05, FDR < 0.1, fold change > 1.5) were detected between the BRSV challenged and control calves. The DE miRNAs were predicted to target 465 genes which were previously found to be DE in bronchial lymph node tissue, between these BRSV challenged and control calves. Of the DE predicted target genes, 455 had fold changes that were inverse to the corresponding DE miRNAs. There were eight enriched pathways among the DE predicted target genes with inverse fold changes to their corresponding DE miRNA including: granulocyte and agranulocyte adhesion and diapedesis, interferon signalling and role of pathogen recognition receptors in recognition of bacteria and viruses. Functions predicted to be increased included: T cell response, apoptosis of leukocytes, immune response of cells and stimulation of cells. Pathogen recognition and proliferation of cytotoxic T cells are vital for the recognition of the virus and its subsequent elimination.