Shandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention, Shandong Agricultural University, 61 Daizong Street, Taian, 271018, Shandong, China.
College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, 210095, China.
Vet Res. 2018 Feb 15;49(1):18. doi: 10.1186/s13567-018-0512-3.
Porcine circovirus type 2 (PCV2) is the primary cause of post-weaning multisystemic wasting syndrome (PMWS) and other PCV-associated diseases. According to our previous RNA-sequencing analysis, the differences in the susceptibility to PCV2 infection depended on the genetic differences between the Laiwu (LW) and Yorkshire × Landrace crossbred (YL) pigs, but the cellular microRNA (miRNA) that are differentially expressed between the LW and YL pigs before and after PCV2 infection remain to be determined. In this study, high-throughput sequencing was performed to determine the abundance and differential expression of miRNA in lung tissues from PCV2-infected and PCV2-uninfected LW and YL pigs. In total, 295 known and 95 novel miRNA were identified, and 23 known and 25 novel miRNA were significantly differentially expressed in the PCV2-infected vs. PCV2-uninfected LW pigs and/or the PCV2-infected vs. PCV2-uninfected YL pigs. The expression levels of ssc-miR-122, ssc-miR-192, ssc-miR-451, ssc-miR-486, and ssc-miR-504 were confirmed by quantitative real-time PCR (qRT-PCR). Analysis of the potential targets of the four up-regulated miRNA (i.e., ssc-miR-122, ssc-miR-192, ssc-miR-451 and ssc-miR-486) identified pathways and genes that may be important for disease resistance. Among the up-regulated miRNA, ssc-miR-122 can repress the protein expression and viral DNA replication of PCV2 and down-regulate the expression of the nuclear factor of activated T-cells 5 (NFAT5) and aminopeptidase puromycin sensitive (NPEPPS) by binding to their 3' untranslated region (3'UTR) in PK15 cells. Therefore, ssc-miR-122 may indirectly suppress PCV2 infection by targeting genes related to the host immune system, such as NFAT5 and NPEPPS.
猪圆环病毒 2 型(PCV2)是断奶后多系统衰竭综合征(PMWS)和其他与 PCV 相关疾病的主要原因。根据我们之前的 RNA 测序分析,对 PCV2 感染的易感性差异取决于莱芜(LW)和约克夏猪×长白猪杂交(YL)猪之间的遗传差异,但 PCV2 感染前后 LW 和 YL 猪之间差异表达的细胞 microRNA(miRNA)仍有待确定。在这项研究中,进行了高通量测序以确定来自 PCV2 感染和未感染的 LW 和 YL 猪的肺组织中 miRNA 的丰度和差异表达。总共鉴定出 295 个已知和 95 个新的 miRNA,并且在 PCV2 感染的 LW 猪与未感染的 LW 猪和/或 PCV2 感染的 YL 猪与未感染的 YL 猪之间,有 23 个已知和 25 个新的 miRNA 显著差异表达。通过定量实时 PCR(qRT-PCR)验证了 ssc-miR-122、ssc-miR-192、ssc-miR-451、ssc-miR-486 和 ssc-miR-504 的表达水平。对四个上调 miRNA(即 ssc-miR-122、ssc-miR-192、ssc-miR-451 和 ssc-miR-486)的潜在靶标进行分析,确定了可能对疾病抵抗力很重要的途径和基因。在上调的 miRNA 中,ssc-miR-122 可以通过结合 PK15 细胞中其 3' 非翻译区(3'UTR)来抑制 PCV2 的蛋白表达和病毒 DNA 复制,并下调核因子活化 T 细胞 5(NFAT5)和氨肽酶 puromycin 敏感(NPEPPS)的表达。因此,ssc-miR-122 可能通过靶向与宿主免疫系统相关的基因(如 NFAT5 和 NPEPPS)间接抑制 PCV2 感染。