Zhao Kui, Li Jida, He Wenqi, Song Deguang, Zhang Ximu, Zhang Di, Zhou Yanlong, Gao Feng
College of Veterinary Medicine, Jilin University, 5333 Xi'an Road, Changchun, 130062, China.
College of Public Hygiene, ZunYi Medical University, 201 Dalian Road, Zunyi, 563003, China.
Virol J. 2017 Jun 15;14(1):114. doi: 10.1186/s12985-017-0781-x.
Viruses interact with host cellular factors to construct a more favourable environment for their efficient replication. Expression of cyclophilin B (CypB), a cellular peptidyl-prolyl cis-trans isomerase (PPIase), was found to be significantly up-regulated. Recently, a number of studies have shown that CypB is important in the replication of several viruses, including Japanese encephalitis virus (JEV), hepatitis C virus (HCV) and human papillomavirus type 16 (HPV 16). However, the function of cellular CypB in ORFV replication has not yet been explored.
Suppression subtractive hybridization (SSH) technique was applied to identify genes differentially expressed in the ORFV-infected MDBK cells at an early phase of infection. Cellular CypB was confirmed to be significantly up-regulated by quantitative reverse transcription-PCR (qRT-PCR) analysis and Western blotting. The role of CypB in ORFV infection was further determined using Cyclosporin A (CsA) and RNA interference (RNAi). Effect of CypB gene silencing on ORFV replication by 50% tissue culture infectious dose (TCID) assay and qRT-PCR detection.
In the present study, CypB was found to be significantly up-regulated in the ORFV-infected MDBK cells at an early phase of infection. Cyclosporin A (CsA) exhibited suppressive effects on ORFV replication through the inhibition of CypB. Silencing of CypB gene inhibited the replication of ORFV in MDBK cells. In conclusion, these data suggest that CypB is critical for the efficient replication of the ORFV genome.
Cellular CypB was confirmed to be significantly up-regulated in the ORFV-infected MDBK cells at an early phase of infection, which could effectively facilitate the replication of ORFV.
病毒与宿主细胞因子相互作用，以构建更有利于其高效复制的环境。细胞肽基脯氨酰顺反异构酶（PPIase）亲环素B（CypB）的表达被发现显著上调。最近，多项研究表明，CypB在包括日本脑炎病毒（JEV）、丙型肝炎病毒（HCV）和16型人乳头瘤病毒（HPV 16）在内的多种病毒的复制中起重要作用。然而，细胞CypB在羊口疮病毒（ORFV）复制中的功能尚未得到探索。
应用抑制性消减杂交（SSH）技术鉴定在感染早期被ORFV感染的MDBK细胞中差异表达的基因。通过定量逆转录PCR（qRT-PCR）分析和蛋白质印迹法证实细胞CypB显著上调。使用环孢素A（CsA）和RNA干扰（RNAi）进一步确定CypB在ORFV感染中的作用。通过50%组织培养感染剂量（TCID）试验和qRT-PCR检测CypB基因沉默对ORFV复制的影响。
在本研究中，发现CypB在感染早期被ORFV感染的MDBK细胞中显著上调。环孢素A（CsA）通过抑制CypB对ORFV复制表现出抑制作用。CypB基因沉默抑制了ORFV在MDBK细胞中的复制。总之，这些数据表明CypB对ORFV基因组的有效复制至关重要。
证实细胞CypB在感染早期被ORFV感染的MDBK细胞中显著上调，这可以有效地促进ORFV的复制。
