Project description:Transcriptional profiling of chicken spleen tissues from Marek's disease virus (MDV) infected and non-infected control individuals from three inbred chicken lines 63, 72, RCSM with varying disease resistance characteristics. Line 63 is highly resistant; line 72 is highly susceptible; and line RCSM has intermediate resistance. Our goal was to investigate the genetic basis of systemic variation in disease resistance in these genetically similar individuals. Three time points were chosen at different stages of disease progression - 5 days post-infection (dpi) - early cytolytic phase; 10 dpi - latent phase; and 21 dpi - late cytolytic phase with 4 replicates at each stage. Two-condition experiment, MDV infected sample vs non-infected control, 3 chicken lines - 63, 72 and RCSM, 3 time points - 5, 10 & 21 dpi. Biological replicates: 4 non-infected control replicates, 4 infected replicates for each line at each time point.
Project description:Transcriptional profiling of chicken spleen tissues from Marek's disease virus (MDV) infected and non-infected control individuals from three inbred chicken lines 63, 72, RCSM with varying disease resistance characteristics. Line 63 is highly resistant; line 72 is highly susceptible; and line RCSM has intermediate resistance. Our goal was to investigate the genetic basis of systemic variation in disease resistance in these genetically similar individuals. Three time points were chosen at different stages of disease progression - 5 days post-infection (dpi) - early cytolytic phase; 10 dpi - latent phase; and 21 dpi - late cytolytic phase with 4 replicates at each stage.
Project description:To investigate specific miRNA expression profiles of Marek's disease virus (MDV)-infected samples, we performed deep sequencing for miRNAs in four small RNA libraries, including MDV-infected tumorous spleen, MD lymphoma from liver, and non-infected spleen and lymphocytes from controls. A total of 7.76x106, 6.36x106, 6.36x106, and 7.60x106 counts were obtained in four libraries, respectively. The sequences were blasted with chicken and MDV genomes and miRBase 16.0 to identify known and novel miRNAs. In total, 187 and 16 known mature miRNAs were identified in the chicken and MDV, respectively. Deep sequencing detected 942 novel chicken miRNA candidates, of which 646 were in tumorous spleen. These results indicate that MDV infection induced new host miRNA candidates and increased diversity of miRNAs. Of 942 miRNA candidates, 276 of 533 were verified by customized microarray, and 17 of them were further confirmed by qPCR. Four samples examined: MDV-infected tumorous spleen, MD lymphoma from liver, Non-infected spleen, Non-infected lymphocytes
Project description:To investigate specific miRNA expression profiles of Marek's disease virus (MDV)-infected samples, we performed deep sequencing for miRNAs in four small RNA libraries, including MDV-infected tumorous spleen, MD lymphoma from liver, and non-infected spleen and lymphocytes from controls. A total of 7.76x106, 6.36x106, 6.36x106, and 7.60x106 counts were obtained in four libraries, respectively. The sequences were blasted with chicken and MDV genomes and miRBase 16.0 to identify known and novel miRNAs. In total, 187 and 16 known mature miRNAs were identified in the chicken and MDV, respectively. Deep sequencing detected 942 novel chicken miRNA candidates, of which 646 were in tumorous spleen. These results indicate that MDV infection induced new host miRNA candidates and increased diversity of miRNAs. Of 942 miRNA candidates, 276 of 533 were verified by customized microarray, and 17 of them were further confirmed by qPCR.
Project description:To gain more insight into the molecular mechanisms of the tumorigenesis of MDV, we used microarrays to screen host and viral miRNAs that were sensitive to infection by MDV. Microarray analysis showed significant differential expression of 79 miRNAs, To determine whether miRNAs were involved in the MDV-induced tumorigenesis, miRNA microarray analysis was performed on GA-infected splenic tumors, GA-infected non-tumorous spleen tissues, and control spleen tissues at 28 dpi.
Project description:Marek's disease virus (MDV) strain GX0101 was the first reported field strain of recombinant gallid herpesvirus type 2 (GaHV-2). However, the splenic proteomics study of MDV strain GX0101 infected chicken was still unclear. In this study, GX0101 was used to infect the chicken spleen in order to analyze the splenic proteomics of the chicken after GX0101 infection.
Project description:To investigate gene expression profiles in susceptible spleens (SS) and resistant spleens (RS) from MDV-infected chickens and non-infected spleens (NS) from controls. Twelve spleens were chosen from three groups (four samples per group) to be used in chicken genome microarray to examine differential gene expression. In total, there were 4317 and 2593 differentially expressed (DE) genes in SS vs. NS and in SS vs. RS, respectively. However, no DE genes were found in RS vs. NS. The expression of twenty-four genes was verified by qPCR. In this study, we found weak, but detectable host immune response still existed in the late neoplastic transformation phase of MDV infection. Toll-like receptor pathway was affected in susceptible spleens compared to non-infected and resistant spleens. IL10 seemed to be exploited by virus to facilitate tumorigenesis. The expression of many members in insulin growth factor system was altered. IGFBP4 and IGFBP7 might be associated with MD lymphoma transformation. The expression profiles in resistant spleens were similar to those in non-infected spleens, which indicated most genes returned to baseline expression levels. Although latent MDV infection persists in resistant chicken for the whole life, it won’t interfere with normal function of genes.
Project description:Expression of known and predicted genes in tissues of Gallus gallus (chicken) pooled from multiple healthy individuals. Two-colour experiments with two different tissues hybridized to each array. Each tissue is arrayed in replicate with dye swaps. Tissues: Bursa of Fabricius, Cerebellum, Cerebral cortex, Eye, Femur with bone marrow, Gallbladder, Gizzard, Heart, Intestine, Kidney, Liver, Lung, Muscle, Ovary, Oviduct, Skin, Spleen, Stomach, Testis, Thymus
Project description:Relative expression levels of mRNAs in chicken cecal epithelia experimentally infected with Eimeria tenella were measured at 4.5 days post-infection. Two weeks old chickens were uninfected (negative control) or were orally inoculated with sporulated oocysts of Eimeria tenella. Cecal epithelia samples were collected from >12 birds in infected or uninfected group at 4.5 d following infections, in which samples from 4 birds were pooled together to form a total 3 biological replicates in each group. Parasite merozoites were also collected from four infected chickens at 5 d after infections. Uninfected control samples, merozoites and infection group samples were selected for RNA extraction and hybridization on Affymetrix microarrays. We used Affymetrix GeneChip chicken genome arrays to detail the chicken cecal epithelia gene expression in the control and E. tenella-infected birds.
Project description:Marek’s disease (MD) is a viral neoplastic disease in chickens caused by the MD virus (MDV). Successful vaccination strategies against MD have resulted in a progressive increase in the virulence of MDV and therefore, the understanding of genetic resistance to MD is considered crucial to the long-term control of the disease. Here we examine whether MDV infection induces changes in the epigenetic status of genes and whether this response is dependent on the host genotype. We generated genome-wide histone 3 lysine 4 (H3K4) and histone 3 lysine 27 (H3K27) trimethylation maps in thymus tissues from chicken lines with varying resistance to MD. Differential chromatin marks were observed on several genes previously implicated in MD such as MX1, MMP2 and CTLA-4 and also on novel genes such as EAF2, IGF2BP1 and GAL. We also detected bivalent domains on transcriptional regulators such as BCL6, CITED2 and EGR1 that have particular functions in immune response. Moreover, novel putative roles for GAL and CITED2 in the mechanism of MD progression were uncovered. We also found tissue-specific effects of MDV infection with certain genes exhibiting differential marks only in spleen. Our results suggest widespread epigenetic changes are induced by MDV infection extent of which is determined by the level of MD resistance of the host. 8 samples examined: 2 histone modifications X 2 chicken lines with varying resistance to MD X 2 groups - infected and control.