Abstract
Avian leukosis virus subgroup J (ALV-J) can cause several different leukemia-like proliferative diseases in the hemopoietic system of chickens. Here, we investigated the transcriptome profiles and miRNA expression profiles of ALV-J-infected and uninfected chicken spleens to identify the genes and miRNAs related to ALV-J invasion. In total, 252 genes and 167 miRNAs were differentially expressed in ALV-J-infected spleens compared to control uninfected spleens. miR-23b expression was up-regulated in ALV-J-infected spleens compared with the control spleens, and transcriptome analysis revealed that the expression of interferon regulatory factor 1 (IRF1) was down-regulated in ALV-J-infected spleens compared to uninfected spleens. A dual-luciferase reporter assay showed that IRF1 was a direct target of miR-23b. miR-23b overexpression significantly (P = 0.0022) decreased IRF1 mRNA levels and repressed IRF1-3′-UTR reporter activity. In vitro experiments revealed that miR-23b overexpression strengthened ALV-J replication, whereas miR-23b loss of function inhibited ALV-J replication. IRF1 overexpression inhibited ALV-J replication, and IRF1 knockdown enhanced ALV-J replication. Moreover, IRF1 overexpression significantly (P = 0.0014) increased IFN-β expression. In conclusion, these results suggested that miR-23b may play an important role in ALV-J replication by targeting IRF1.
Highlights
Mature microRNAs are single-stranded non-coding small RNAs of 21-25 nucleotide lengths that typically reduce the translation and stability of mRNAs. miRNA deregulation has been shown to play pivotal roles in tumorigenesis and progression via the up-regulation of oncogenes and the silencing of tumor suppressor genes, respectively[9,10]
These same DNA samples did not produce any specific products during PCR with primers used for the detection of other viruses, including exogenous ALVs (Fig. 1C), Marek’s disease virus (MDV) (Fig. 1D) and reticuloendotheliosis virus (REV) (Fig. 1E)
immunofluorescence assay (IFA) indicated positive results for the ELISA-positive samples (WRR1+, WRR2+, WRR3+) (Fig. 1F) but negative results for the ELISA-negative samples (WRR1−, WRR2−, WRR3−), confirming that the samples WRR1+, WRR2+, and WRR3+were infected with Avian leukosis virus subgroup J (ALV-J), whereas the samples WRR1−, WRR2−, and WRR3− were not infected
Summary
Mature microRNAs (miRNAs) are single-stranded non-coding small RNAs of 21-25 nucleotide (nt) lengths that typically reduce the translation and stability of mRNAs. miRNA deregulation has been shown to play pivotal roles in tumorigenesis and progression via the up-regulation of oncogenes and the silencing of tumor suppressor genes, respectively[9,10]. The objective of the present study was to investigate the genetic basis of host resistance against ALV-J and to identify key miRNAs and target genes responsible for the host anti-ALV-J response. The transcriptome profiles and miRNA expression profiles of ALV-J-infected and uninfected chicken spleens were scanned to identify genes and miRNAs related to ALV-J invasion. Targets of these differentially expressed miRNAs were predicted, and differentially expressed genes (DEGs) that are the targets of the differentially expressed miRNAs were selected. In vitro experiments were performed to identify candidate genes and miRNAs involved in host anti-ALV-J mechanisms
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