Abstract
BackgroundViruses 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.MethodsSuppression 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 (TCID50) assay and qRT-PCR detection.ResultsIn 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.ConclusionsCellular 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.
Highlights
Viruses interact with host cellular factors to construct a more favourable environment for their efficient replication
cyclophilin B (CypB) was found to be significantly up-regulated in the Orf virus (ORFV)-infected Madin―Darby bovine kidney (MDBK) cells by Suppression subtractive hybridization (SSH) technique In order to explore host genes inovolved in the process of ORFV invasion, the differentially expressed genes from ORFV-infected MDBK cells at an early phase of infection were identified using SSH technique
Among of all the expressed sequence tags (ESTs), 60 ESTs came from the forward SSH library and 21 ESTs came from the reverse SSH library
Summary
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. 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). Cyclophilins (Cyp) comprise a family of peptidyl-prolyl cis/trans isomerases, which are originally discovered as a cellular factor with high affinity for the immunosuppressant CsA [4, 5]. CypB functions in various cellular processes, including transcriptional regulation, protein secretion, immune response and apoptosis [8,9,10]. Some researchers have shown that many viruses such as Japanese encephalitis virus (JEV), hepatitis C virus (HCV) and human papillomavirus type 16 (HPV 16)
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