Project description:Infectious bursal disease virus (IBDV) causes a highly contagious, immunosuppressive disease in chickens. The virus mainly infects immature B lymphocytes in the bursa of Fabricius (BF). Chicken B cell line DT40, an avian leukosis virus-induced B cell line, supports very virulent IBDV (vvIBDV) infection in vitro and thereby serves as a good model for investigating the infection and pathogenesis of this virus. However, a transcriptome-wide understanding of the interaction between vvIBDV and B cells has not yet been achieved. This study aimed to employ time-course DNA microarrays to investigate gene expression patterns in DT40 cells after infection with vvIBDV strain LX.
Project description:Purpose: To explore the interaction between host and IBDV, RNA-Seq was applied to analyse the transcriptional profiles of the responses of chickens' bursas of Fabricius in the early stage of IBDV infection. Method: Eighteen SPF white leghorn chickens were randomly divided into two groups with 9 chickens for each group: the mock group (the healthy group) and the IBDV-inoculated group (the infection group). Chickens from the infection group were inoculated with 0.1 mL of 103 EID50 IBDV CJ801 stock through eye-nose drops. The chickens from the mock group were kept in a separate isolator and mock challenged with PBS. On days 1, 3 and 7 after infection, 3 chickens from each group were killed for bursa collection. Each bursa was immediately put into liquid nitrogen and then stored in 80 ℃ refrigerator. RNA sequencing was performed with total RNA from bursae from each group at the first two time points and completed by a commercial company. Results: The results displayed that a total of 15546 genes were identified in the chicken bursa libraries. Among the annotated genes, there were 2006 and 4668 differentially expressed genes in the infection group compared with the mock group on day 1 and day 3 post inoculation (1 and 3 dpi), respectively. Moreover, there were 676 common up-regulated and 83 common down-regulated genes in the bursae taken from the chickens infected with IBDV on both 1 and 3 dpi. Meanwhile, there were also some characteristic differentially expressed genes on 1 and 3 dpi. On day 1 after inoculation with IBDV, host responses mainly displayed immune response processes, while metabolic pathways played an important role on day three post infection. Six genes were confirmed by quantitative reverse transcription-PCR. Conclusions: In conclusion, the differential gene expression profile demonstrated with RNA-Seq might offer a better understanding of the molecular interactions between host and IBDV during the early stage of infection.
Project description:Background: Avian infectious bursal disease virus (IBDV) is a major poultry disease which leads to significant losses of poultry industry. Dendritic cells (DCs), the only bridge communicated the innate and acquired immunity, have the most important antigen presenting ability and can significantly influence the pathogenicity of viruses. To understand the interaction between IBDV and DCs, microarray was used to analyze the response of DCs infected by IBDV. Results: Results showed that IBDV infection induced 479 up-regulated and 466 down-regulated mRNAs in chicken DCs. GO terms analysis suggested that transcription from RNA polymerase II promoter and RNA biosynthetic process were mainly enriched, whilst pathway analyses suggested that oxidative phosphorylation, T cell receptor and IL-17 signaling pathway might activated by IBDV infection. Moreover, we detected the microRNA (miRNA) and long non-coding RNA (lncRNA) alterations in IBDV-infected chicken DCs. Results identified 18 significant up- or down- regulated miRNAs and 441 significant up- or down-regulated lncRNAs in IBDV-stimulated DCs. Furthermore, we constructed 42 TF (transcription factors)-miRNA-mRNA interactions involving 1 TF, 3 miRNAs and 42 mRNAs in IBDV-stimulated DCs. Finally, we predicted the target genes of different expressed lncRNAs and constructed lncRNA-mRNA regulatory networks. Conclusions: Altogether, our research suggested a mechanism to explain how IBDV infection triggered an effective immune response in chicken DCs.
Project description:Infectious bursal disease virus (IBDV) is a highly contagious dsRNA virus (Birnaviridae) which causes immuno-suppression in chickens. Although largely controlled by vaccination, new, virulent strains of the virus mean that infectious bursal disease (IBD or ëGumboroí disease) still remains a threat to the poultry industry. The virus infects dividing IgM+ B-lymphocytes and the main site of viral replication is the bursa of Fabricius where B cells are produced. Infection is spread orally via contaminated feed and water. IBDV affects young birds, with the disease usually being diagnosed in 3-6 week old birds. Younger birds do not show clinical signs but are immuno-suppressed. Symptoms include anorexia, depression, diahorrea, ruffled feathers, immuno-suppression and bursal lesions. The disease peaks between 2-5 days post infection and is practically cleared by day 7. Mortality is variable but can be up to around 70% with very virulent strains of the disease. Even if birds survive, the resulting immuno-suppression and effect on egg production in layer birds is significant. Being able to breed commercial lines of birds for enhanced genetic resistance to IBDV is an obvious goal in the fight against the disease. Three-week-old chicks were inoculated with virus via an intra-nasal route and tissue samples were collected at 2, 3 and 4 days post-inoculation. Bursa and spleen tissues were examined from control and infected birds at 2, 3 and 4 days post-infection in birds known to be either susceptible or resistant to the virus. As well as understanding the host immune response to IBDV, we are interested in identifying genes involved in disease resistance and so we have analysed the gene expression profiles at these times, when the innate immune response is active. We assume that genes underlying resistance will be involved at this early stage of the host immune response.
Project description:Infectious bursal disease virus (IBDV) is the pathogenic agent of infectious bursal disease (IBD). Scine it was observed in 1957, IBD spread worldwidely in the chicken flocks, is a important immunosuppressive disease and an threat to poultry industry. Although many studies have be done about IBDV, interaction of IBDV infection and IBDV-encoding genes to host cell gene expression are little known. In this study, the LongSAGE library of Vero-cell, IBDV- infected vero cell, Vero-cell transfected with IBDV-VP5 gene, Vero-cell transfected with IBDV A frament and Vero-cell transfected with IBDV VP243 frament were obtained. We got 96,213 gene tags (17 nucleotides), which represented 24,475 transcripts. Keywords: Transcripts of different state vero-cell
Project description:Chickens known to be susceptible to IBDV were infected with the virus and resulting gene expression at 4dpi was compared to that of uninfected chickens
Project description:Infectious bursal disease virus (IBDV) is the pathogenic agent of infectious bursal disease (IBD). Scine it was observed in 1957, IBD spread worldwidely in the chicken flocks, is a important immunosuppressive disease and an threat to poultry industry. Although many studies have be done about IBDV, interaction of IBDV infection and IBDV-encoding genes to host cell gene expression are little known. In this study, the LongSAGE library of Vero-cell, IBDV- infected vero cell, Vero-cell transfected with IBDV-VP5 gene, Vero-cell transfected with IBDV A frament and Vero-cell transfected with IBDV VP243 frament were obtained. We got 96,213 gene tags (17 nucleotides), which represented 24,475 transcripts. Keywords: Transcripts of different state vero-cell 1.Cloning of the full-length genomic A-segment, VP5 ORF cDNA, VP243 ORF cDNA of IBDV 2.Establishing cloned Vero cell lines expressing VP5, VP243 and A fragment of IBDV 3.Construction of Long-SAGE libraries 4. Sequencing
Project description:Viral diseases pose major threats to humans and other animals, including the billions of chickens that are an important food source as well as a public health concern due to zoonotic pathogens. Unlike humans and other typical mammals, the major histocompatibility complex (MHC) of chickens can confer decisive resistance or susceptibility to many viruses. Examples are Marek's disease virus (MDV) and Infectious bursal disease virus (IBDV). We used a new in vitro infection system and immunopeptidomics to identify peptides presented to T lymphocytes via classical MHC class II molecules.
Project description:Infectious bursal disease virus (IBDV) belongs to the family Birnaviridae and is economically important to the poultry industry worldwide. IBDV infects B cells in the bursa of Fabricius (BF), which can cause severe immunosuppression and mortality in young chickens. Earlier studies have shown that strains of IBDV lose their virulence potential after serial passage in non-B lymphoid cells, for reasons that are poorly understood. This study aimed to investigate the gene expression profiles of one cell-culture adapted attenuated IBDV strain (D78) and one very virulent IBDV strain (UK661) in chicken primary B cells cultured ex vivo from the bursa of Fabricius. The viruses were studied in B cells over 48h and their gene expression was initially evaluated with qPCR. The mRNA was isolated from the cells at 18 hours post-infection and screened with Affymetrix microarrays in triplicate. The study included mock controls which were conducted in triplicate.
Project description:Infectious Bursal Disease Virus (IBDV) causes highly contagious, immunosuppressive disease that leads to high mortality in young chickens. Chicks of a F2 generation of two lines divergently selected for high (HH) or low (LL) antibody (Ab) response to Escherichia coli vaccination, were challenged with virulent IBDV. Viral load in infected bursae was used to determine resistant (R) and susceptible (S) birds. By using a 13K chicken cDNA microarray, and pooled spleens of R, S and non-challenged, control (C) chicks, several genes were identified with differential expression associated with host resistance to IBDV. These genes were also subjected to RT-PCR on individual samples to verify the results obtained from microarrays. The major finding was the co-upregulation of 7 genes – coding for Ets2, H963, RGS1, ABIN-2, CREM/ICER, DUSP1 and CXCR4 – in several R, but not S or C, individuals, and characterized by very high correlations of expression levels. Resistance also generally coincided with reduced transcript levels of acute-phase serum amyloid A (A-SAA) and increased levels of IL-8. Based on reported functions of these genes, our findings suggest that resistance was mediated by the activation of specific cellular mechanisms, indicated by increased activity of splenic macrophages and T-lymphocytes 3d post-challenge. Early and intense innate responses may enhance the formation and activity of germinal centers in the spleen of resistant birds, and the transition to acquired cellular response. The migration of activated cells towards the bursa is presumably important for resistance to occur. Keywords: host-resistance analysis