Project description:B lymphocytes from the chicken Bursa of Fabricius were isolated and cultured in the presence of CD40L. Cells were infected with a GFP fluorescent reporter virus of the very virulent Marek's disease virus (vvMDV) strain RB-1B (RB-1B_UL47-GFP, see doi: 10.1073/pnas.1424420112). After 24h viable infected (GFPpositive) and uninfected (GFPnegative) B-cells were sort purified from these cultures. Viable B cells, purified from uninfected cultures served as control. 3x10^6 cells per sample were subjected to RNA isolation and microarray analysis. Goal of the experiment was to elucidate the reaction of chicken B cells, primary target cells of MDV upon infection with this virus.
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: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:Marek's disease (MD), induced by Marek's disease virus (MDV), is a lymphotropic neoplastic disease and causes huge economic losses to the poultry industry. Non-coding RNAs play important regulatory roles in disease pathogenesis. To investigate host miRNA expression profile, RNA sequencing was performed in tumorous spleens (TS), spleens from the survivors (SS) without any lesion after MDV infection, and noninfected chicken spleens (NS).
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:Marek's disease (MD), induced by Marek's disease virus (MDV), is a lymphotropic neoplastic disease and causes huge economic losses to the poultry industry. Non-coding RNAs play important regulatory roles in disease pathogenesis. To investigate host circRNA and miRNA expression profile, RNA sequencing was performed in tumorous spleens (TS), spleens from the survivors (SS) without any lesion after MDV infection, and noninfected chicken spleens (NS).
Project description:We infected chickens from two genetic lines, one bred to be resistant to Marek's disease and one bred to be susceptible, with the Marek's disease virus. We performed single-cell RNA sequencing of the spleens of these chickens along with age-matched uninfected controls from both lines.
Project description:Marek's disease virus (MDV) is a highly contagious oncogenic herpes virus. Classed as an alpha herpes virus due to its DNA sequence and genome organization, its biological properties are more similar to those of gamma herpes viruses. MDV causes an initial cytolytic infection in B lymphocytes (3-6dpi) followed by infection of CD4+ T cells (at around 7dpi) where the virus becomes latent (for around two weeks) and then goes on to produce lymphoid tumours in the skin, nerves and internal organs. Early symptoms in susceptible birds include lameness, paralysis, loss of appetite, depression, blindness and immuno-suppression. The route of infection is usually respiratory and the disease is highly contagious, being spread by infective feather-follicle dander. Although birds may survive MDV infection, the resultant immuno-suppression leaves the birds highly susceptible to other infections. MDV infection can thus be associated with high mortality rates and, in turn, large economic losses throughout the worldï¾Âs poultry industry. Three-week-old chicks were inoculated with virus (RB1B) via an intra-gastric route and tissue samples were collected at 2, 3 and 4 days post-inoculation. Spleen and thymus tissue 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 MDV, 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.