Cheng Feng, Wang Hui, Zhou Lei, Lan Ganqiu, Yang Hanchun, Wang Lixian, Wang Ligang, Liang Jing
College of Animal Science and Technology, Guangxi University, Nanning 530004, China.
Key Laboratory of Farm Animal Genetic Resources and Germplasm Innovation of Ministry of Agriculture of China, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China.
Animals (Basel). 2023 Feb 28;13(5):876. doi: 10.3390/ani13050876.
Exosomes are biological vesicles secreted and released by cells that act as mediators of intercellular communication and play a unique role in virus infection, antigen presentation, and suppression/promotion of body immunity. Porcine reproductive and respiratory syndrome virus (PRRSV) is one of the most damaging pathogens in the pig industry and can cause reproductive disorders in sows, respiratory diseases in pigs, reduced growth performance, and other diseases leading to pig mortality. In this study, we used the PRRSV NADC30-like CHsx1401 strain to artificially infect 42-day-old pigs and isolate serum exosomes. Based on high-throughput sequencing technology, 305 miRNAs were identified in serum exosomes before and after infection, among which 33 miRNAs were significantly differentially expressed between groups (13 relatively upregulated and 20 relatively downregulated). Sequence conservation analysis of the CHsx1401 genome identified 8 conserved regions, of which a total of 16 differentially expressed (DE) miRNAs were predicted to bind to the conserved region closest to the 3' UTR of the CHsx1401 genome, including 5 DE miRNAs capable of binding to the CHsx1401 3' UTR (ssc-miR-34c, ssc-miR-375, ssc-miR-378, ssc-miR-486, ssc-miR-6529). Further analysis revealed that the target genes of differentially expressed miRNAs were widely involved in exosomal function-related and innate immunity-related signaling pathways, and 18 DE miRNAs (ssc-miR-4331-3p, ssc-miR-744, ssc-miR-320, ssc-miR-10b, ssc-miR-124a, ssc-miR-128, etc.) associated with PRRSV infection and immunity were screened as potential functional molecules involved in the regulation of PRRSV virus infection by exosomes.
外泌体是细胞分泌并释放的生物囊泡,作为细胞间通讯的介质,在病毒感染、抗原呈递以及机体免疫的抑制/促进过程中发挥独特作用。猪繁殖与呼吸综合征病毒(PRRSV)是养猪业中危害最大的病原体之一,可导致母猪繁殖障碍、猪呼吸道疾病、生长性能下降以及其他导致猪死亡的疾病。在本研究中,我们使用PRRSV NADC30样CHsx1401毒株人工感染42日龄仔猪并分离血清外泌体。基于高通量测序技术,在感染前后的血清外泌体中鉴定出305个miRNA,其中33个miRNA在组间存在显著差异表达(13个相对上调,20个相对下调)。对CHsx1401基因组进行序列保守性分析,鉴定出8个保守区域,其中共有16个差异表达(DE)miRNA被预测与最接近CHsx1401基因组3'UTR的保守区域结合,包括5个能够与CHsx1401 3'UTR结合的DE miRNA(ssc-miR-34c、ssc-miR-375、ssc-miR-378、ssc-miR-486、ssc-miR-6529)。进一步分析表明,差异表达miRNA的靶基因广泛参与外泌体功能相关和先天免疫相关信号通路,筛选出18个与PRRSV感染和免疫相关的DE miRNA(ssc-miR-4331-3p、ssc-miR-744、ssc-miR-320、ssc-miR-10b、ssc-miR-124a、ssc-miR-128等)作为参与外泌体调控PRRSV病毒感染的潜在功能分子。