Project description:Subgroup J avian leukemia is a type of oncology infectious disease caused by Subtype J of avian leukosis virus (ALV-J). It mainly encroaches on bone marrow cells, and metastasizes to liver, kidney, splenic ellipsoids and other organs, leading to myeloid leukosis (ML) and other malignancies, resulting in significant economic losses. microRNA play important roles in oncology infectious diseases. We used miRNA microarray analysis to detail the relationship of aberrant microRNAs and chicken ALV-J leukemia, and to try to find the potential diagnostic and therapeutic target for infections of subtype J of leukemia.
Project description:To survey avian leukosis virus subgroup J (ALV-J) integration in myeloid leukosis (ML) of chicken, we developed an ALV-J insertional identification platform based on hybrid-capture target enrichment and next-generation sequencing (NGS). In addition, we used gene expression profiling and bioinformatics to associate integration sites to transcriptional activity and to genetic features of the tumor cell genome.
Project description:Subgroup J avian leukemia is a type of oncology infectious disease caused by Subtype J of avian leukosis virus (ALV-J). It mainly encroaches on bone marrow cells, and metastasizes to liver, kidney, splenic ellipsoids and other organs, leading to myeloid leukosis (ML) and other malignancies, resulting in significant economic losses. microRNA play important roles in oncology infectious diseases. We used miRNA microarray analysis to detail the relationship of aberrant microRNAs and chicken ALV-J leukemia, and to try to find the potential diagnostic and therapeutic target for infections of subtype J of leukemia. ALV-J-infected and non-infected ten-week-old chicken liver tissues were sampled for the array assay. A1, A2, and A3 are the ALV-J infection group samples, and DA1, DA2, and DA3 are the control samples. 623 mature miRNA sequences were assembled and integrated into the LC miRNA microarray design, and different miRNA expressions were measured on the 7000HT Fast Real-Time PCR system.
Project description:To survey avian leukosis virus subgroup J (ALV-J) integration in myeloid leukosis (ML) of chicken, we developed an ALV-J insertional identification platform based on hybrid-capture target enrichment and next-generation sequencing (NGS). In addition, we used gene expression profiling and bioinformatics to associate integration sites to transcriptional activity and to genetic features of the tumor cell genome. We selected six cases of ALV-J positive and diagnosed as ML for integration sites identify from commercial broiler breeder flocks in Guangdong Province of China between November 2011 and March 2012. All tumors were diagnosed on the basis of characteristic gross and microscopic lesions. Furthermore, PCR tests on the genomic DNA of tissues and virus isolation assay only showed ALV-J-specific positive results in previously study. We randomly chose 4 independent liver samples from the six cases for gene expression profile analysis. And 3 ALV-negative tissue samples from specific-pathogen-free (SPF) chickens at the same age were use as negative controls. Thus a total of 7 samples were hybridized, three representing control.
Project description:PIWI-interacting RNAs (piRNAs) protect the germ line by targeting transposable elements (TEs) through base-pair complementarity. We do not know how piRNAs co-evolve with TEs in chickens. Here we reported that all active TEs in the chicken germ line are targeted by piRNAs, and as TEs lose their activity, the corresponding piRNAs erode away. We observed de novo piRNA birth as host responds to a recent retroviral invasion. Avian leukosis virus (ALV) has endogenized prior to chicken domestication, remains infectious, and threatens poultry industry. Domestic fowl produced piRNAs targeting ALV from a genomic locus that was known to render its host ALV resistant. This genomic locus does not produce piRNAs in undomesticated wild chickens. Our findings uncover rapid piRNA evolution reflecting contemporary TE activity, identify a new piRNA acquisition modality by activating a pre-existing genomic locus, and extend piRNA defense roles to include the period when endogenous retroviruses are still infectious.
Project description:Background:Dendritic cells (DCs), have the most important antigen presenting ability and played an irreplaceable role in recognizing and clearing virus. Antiviral responses must rapidly defend against infection while minimizing inflammatory damage, but the mechanisms that regulate the magnitude of response within an infected cell are not well understood. MicroRNA, small non-coding RNAs, that can regulate dendritic cells to inhibit the infection and replication of avian influenza virus. Here, we global analyses how avian DCs response to H9N2 avian influenza virus (AIV) and provide a potential mechanism of how avian microRNA defending H9N2 AIV replication. Results: Here, we global analyses how avian DCs response to H9N2 avian influenza virus (AIV) and provide a potential mechanism of how avian microRNA defending H9N2 AIV replication. First, we found that both active and inactive H9N2 AIV enhance the ability of DCs to present antigens and activate T lymphocytes. Next, total microarray analyses suggested that H9N2 AIV stimulation involved in protein localization, nucleotide binding and leukocyte transendothelial migration and MAPK signal pathways. Moreover, we construct 551 transcription factor (TF)-microRNA-mRNA loops based on the above analyses. Furthermore, we found that HA fragment could not activate DCs, while truncated HA highly increased the immune function of DCs by activating ERK and STAT3 signal pathway. Last, our insight research not only gained that gga-miR1644 might target to MBNL2 to enhanced avian DCs in inhibiting virus replication, but also suggested that gga-miR6675 target to the NLS of PB1 to trigger the silencing of PB1 genes and lead to inhibition of H9N2 avian influenza viral replication. All together, our innovative research will shed new light on the roles of avian microRNA in evoking avian DCs and inhibiting virus replication, which will suggest new strategies to combat avian influenza virus.
Project description:Purpose: The goals of this study are to investigate the differentially expressed miRNAs between ALV-J infected (WRR+) and uninfected (WRR-)chickens spleens by Illumina deep sequencing. Methods: 140-day-old female chickens of White Recessive Rock (WRR) were confirmed as J subgroup avian leukosis virus (ALV-J) infection. Total RNA from three ALV-J-infected spleens (designated: WRR1+, WRR2+, WRR3+) and three uninfected normal spleen samples (designated: WRR1-, WRR2-, WRR3-) was isolated by TRIzol following the manufacturer’s instruction (Invitrogen, CA, USA). RNA samples of three individuals within each group were pooled with equal amounts, and then were subjected to Illumina deep sequencing by Illumina Hiseq 2500. Results: After raw data filtered, 12,150,275 and 15,227,930 reads of 18-32 bp, representing 569,847 and 543,062 unique sequences, were obtained for WRR- and WRR+ libraries, respectively. Through blasting with the chicken reference genome, 360,180 WRR- sequences and 327,391 WRR+ sequences, which accounted for more than 60% of the unique sequences, were perfectly matched.To analyze the miRNA detection efficiency of Illimuna deep sequencing, all the clean reads were blasted with the Rfam data base 10.1, annotated and then removed rRNA, tRNA, snoRNA and other snRNAs. The annotation results revealed that miRNAs accounted for more than 68% of all clean reads in the WRR− and WRR+ libraries. In this study, a total of 476 miRNAs were identified after compared the unique sequences against the chicken miRNAs precursors in miRBase 18.0. Base on unique sequences matched counts, 167 differential expression miRNAs were identified by DEGseq package using Benjamini-q-value of 0.001 as a cut-off. In ALV-J infected spleens, 83 miRNAs showed up-regulated expression and 84 were down-regulated when compared to uninfected samples. Conclusions: Our study represents the first time to analysis of miRNA Expression in Spleen of J Subgroup Avian Leukosis Virus (ALV-J) Infected (WRR+) and Uninfected (WRR-) Broilers. A total of 167 miRNAs were found to be differentially expressed in ALV-J infected spleens when compared to uninfected chickens. These miRNAs can be considered as candidates for further study ALV-J invasion.
Project description:A novel avian-origin H7N9 influenza A virus (IAV) emerged in China in early 2013 causing mild to lethal human respiratory infections. H7N9 originated from multiple reassortment events between avian viruses and carries genetic markers of human adaptation. Determining whether H7N9 induces a host-response closer to human or avian IAV is important to better characterize this emerging virus. Here we compared the human lung epithelial cell response to infection with A/Anhui/01/13 (H7N9) or highly pathogenic avian-origin H5N1, H7N7, or human seasonal H3N2 IAV.