College of Veterinary Medicine, Shandong Agricultural University, Tai'an, 271018, China.
The Pirbright Institute & UK-China Centre of Excellence on Avian Disease Research, Pirbright, Ash Road, Guildford, Surrey, GU24 0NF, UK.
Retrovirology. 2018 Jul 3;15(1):45. doi: 10.1186/s12977-018-0427-0.
Co-infection with avian leukosis virus subgroup J and reticuloendotheliosis virus induces synergistic pathogenic effects and increases mortality. However, the role of exosomal miRNAs in the molecular mechanism of the synergistic infection of the two viruses remains unknown.
In this study, exosomal RNAs from CEF cells infected with ALV-J, REV or both at the optimal synergistic infection time were analysed by Illumina RNA deep sequencing. A total of 54 (23 upregulated and 31 downregulated) and 16 (7 upregulated and 9 downregulated) miRNAs were identified by comparing co-infection with two viruses, single-infected ALV-J and REV, respectively. Moreover, five key miRNAs, including miR-184-3p, miR-146a-3p, miR-146a-5p, miR-3538 and miR-155, were validated in both exosomes and CEF cells by qRT-PCR. GO annotation and KEGG pathway analysis of the miRNA target genes showed that the five differentially expressed miRNAs participated in virus-vector interaction, oxidative phosphorylation, energy metabolism and cell growth.
We demonstrated that REV and ALV-J synergistically increased the accumulation of exosomal miRNAs, which sheds light on the synergistic molecular mechanism of ALV-J and REV.
禽白血病病毒亚群 J 和网状内皮组织增生症病毒的共同感染会引起协同的致病作用，并增加死亡率。然而，外泌体 miRNA 在这两种病毒协同感染的分子机制中的作用尚不清楚。
在这项研究中，通过 Illumina RNA 深度测序分析了在最佳协同感染时间用 ALV-J、REV 或两者感染的 CEF 细胞中的外泌体 RNA。通过比较两种病毒的共感染、单一感染的 ALV-J 和 REV，分别鉴定出 54 个（23 个上调和 31 个下调）和 16 个（7 个上调和 9 个下调）miRNA。此外，通过 qRT-PCR 在两种外泌体和 CEF 细胞中验证了 5 个关键 miRNA，包括 miR-184-3p、miR-146a-3p、miR-146a-5p、miR-3538 和 miR-155。miRNA 靶基因的 GO 注释和 KEGG 通路分析表明，这 5 个差异表达的 miRNA 参与了病毒-载体相互作用、氧化磷酸化、能量代谢和细胞生长。
我们证明了 REV 和 ALV-J 协同增加了外泌体 miRNA 的积累，这为 ALV-J 和 REV 的协同分子机制提供了启示。
