RNA-seq for splenic gene expression in response to Newcastle disease virus challenge in two chicken lines with different disease resistance
ABSTRACT: In order to have a better understanding about gene expression response to NDV in chicken spleen, and also to unravel genetic regulation related to resistance to NDV, gene expression in spleen of two chicken lines [Fayoumi (resistant line) and Leghorn (susceptible line)] with different resistance to infectious diseases were investigated. Each line was divided into two groups (3-4 chickens/group) that are respectively treated by NDV(200 ul of 107 EID50%) and Phosphate-buffered saline (PBS) through nasal and ocular inoculation routes at 21 days post hatch. Gene expression in spleen was then detected by RNA-seq at 2 and 6 day post inoculation (dpi).
Project description:Genotype VIId NDV is characterized by severe tissue damage in chicken lymphoid organs compared to other virulent strains. However, biological basis of this unusual pathological phenotype is unknown. Host response is associated with pathogenicity of Newcastle Disease Virus (NDV). We aim to determine the contribution of host response to the severe tissue destruction in the lymphoid system caused by genotype VIId NDV. We used microarray analysis to evaluate the global transcriptional response in the spleen of chickens infected with genotype VIId NDV strain JS5/05 and genotype IV NDV Herts/33. Chickens were inoculated with JS5/05 or Herts/33 or mock-infected. At day 2 post infection, spleens were isolated from three chickens per group for RNA extraction and hybridization on Affymetrix microarrays. Samples were named as follows: JS5/05 (I4_1_NS,I4_2_NS,I4_3_NS), Herts/33 (H1_NS,H2_NS, H3_NS), control (C1_NS, C2_NS, C3_NS).
Project description:Chicken anaemia virus (CAV) causes severe anaemia and immunosuppression in young chickens. Such effects reduce the efficiency of routine vaccinations while aggravating the effects of other pathogens in chicken populations. So far, the host responses to CAV have not been studied in vivo on a whole genome-wide scale. In this study, we compared gene expression profiles of chicken tissues (thymus, bone marrow, bursa of Fabricius and spleen-in vivo) from SPF chickens at 14-day-old following intramuscular inoculation at day-old. A chicken specific-immune microarray (5k) developed at Roslin Institute was used throughout the study. Statistically significant gene expression changes occurred mainly in the thymus (in vivo).
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:A unique, lymphoid tissue located adjacent to the eye of the chicken, the Harderian gland is assumed to have critical immune functions, but prior to the current study, its transcriptome had never been analyzed. This study aimed to characterize differential responses between the relatively resistant (Fayoumi) and susceptible (Leghorn) chicken lines when challenged with a high titered lentogenic Newcastle Disease Virus (NDV) strain via the eyes and nares at three weeks of age. The Harderian gland was collected from challenged and non-challenged birds from each line at 2, 6, and 10 days-post-infection (dpi) (n=47). High quality RNA was isolated, used for cDNA library construction, and sequenced on the HiSeq2500.
Project description:This study aimed to characterize differential responses between the relatively resistant (Fayoumi) and susceptible (Leghorn) chicken lines when challenged with a high titered lentogenic Newcastle Disease Virus (NDV) strain via the eyes and nostrils at three weeks of age. The trachea epithelial cells were collected from challenged and non-challenged birds from each line at 2, 6, and 10 days-post-infection (dpi). High quality RNA was isolated, used for cDNA library construction, and sequenced on the HiSeq2500.
Project description:This study aimed to characterize differential responses between the relatively resistant (Fayoumi) and susceptible (Leghorn) chicken lines when challenged with a high titered lentogenic Newcastle Disease Virus (NDV) strain via the eyes and nostrils at three weeks of age. The lung was collected from challenged and non-challenged birds from each line at 2, 6, and 10 days-post-infection (dpi). High quality RNA was isolated, used for cDNA library construction, and sequenced on the HiSeq2500.
Project description:Chicken 60-mer oligonucleotide microarray, including 39854 cDNA and ESTs, entire Marek’s disease virus and avian influenza virus genomes, and 150 chicken microRNAs, was developed. Cecal tonsil, ileum, liver and spleen from 6 chickens were selected for hybridization to validate the microarray performance. There are 2886, 2886, 2660, 358, 3208 3355, and 3710 genes significantly expressed between liver and spleen, spleen and cecal tonsil, cecal tonsil and ileum, liver and cecal tonsil, liver and ileum, spleen and ileum at the P<10-7. Number of tissue specific genes for cecal tonsil, ileum, liver and spleen was 95, 71, 535, and 108, respectively with p < 10-7. More than 95% of spots had high SNR (>10). Keywords: characteristics of newly developed microarray using different normal tissue Loop design was carried on for all of tissue samples from the six chickens. Samples of four tissues from a chicken were used in each loop. The order of the tissues in each loop was changed so that all pairs of tissues were combined on an array with an equal number of times. Dye swap was used so that each tissue was measured an equal number of times with each dye. Data from 12 measurements for each tissue were collected, in total, 48 measurements from 24 arrays.
Project description:Cellular proteins in central nervous system involved in avian neurotropic virus infection remains completely unknown. To investigate host gene expression profile in NDV infected SPF chicken brains, The microarray initial analysis was performed at LC-Bio (Hangzhou, China). A 44K Agilent chicken whole genome chip (43,803 probes) (Agilent Technologies, USA) was used for gene microarray analysis from F48E9-, LaSota-infected and mock-infected brains through intraocular-nasal routes.The chicken brains were collected at 5 day post infection. The significance analysis was used to evaluate the differences in gene expression. P and fold change (FC) values represent the alteration tendency of gene expression between experimental and control groups. The genes (FC>2) were input in Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis for DEGs. Overall design: F48E9-, LaSota-infected and PBS (control)-infected brains through intraocular-nasal routes in 100000 pfu/100μl/chicken were collected at 5 day post infection.The data from F48E9- and LaSota-infected brains were compared to mock (PBS)-infection.
Project description:Three cDNA libraries of mixed visceral tissues from F48E9, La Sota, or uninfected chicken embryos were constructed, and small-RNA deep sequencing was conducted to detect the expression levels of small-RNAs. Intergroup comparisons were used to identify changes in miRNA expression caused by NDV infection. La Sota affected the expression of 61 miRNAs (36 upregulated and 25 downregulated) at 36 hpi, and F48E9 infection altered the expression levels of 66 miRNAs (33 upregulated and 31 downregulated). Overall design: Using high-throughput sequencing, revealed miRNA expression differences of SPF chick embryos infected different virulence NDVs.
Project description:White Leghorn chicken eggs were incubated for 18 days and dissected. Brain, breast muscle, bursa Fabricii, heart, kidney, liver, lung, ovary, spleen, and testicle tissues were sampled.