Project description:Mycobacterium bovis is an intracellular pathogen that causes tuberculosis in cattle. Following infection, the pathogen resides and persists inside host macrophages by subverting host immune responses via a diverse range of mechanisms. Here, a high-density bovine microarray platform was used to examine the bovine monocyte-derived macrophage transcriptome response to M. bovis infection relative to infection with the attenuated vaccine strain, M. bovis Bacille CalmetteM-bM-^@M-^SGuM-CM-)rin. Differentially expressed genes were identified (adjusted P-value M-bM-^IM-$ 0.01) and interaction networks generated across an infection time course of 2, 6 and 24 h. The largest number of biological interactions was observed in the 24 h network, which exhibited small-worldscale-free network properties. The 24 h network featured a small number of key hub and bottleneck gene nodes, including IKBKE, MYC, NFKB1 and EGR1 that differentiated the macrophage response to virulent and attenuated M. bovis strains, possibly via the modulation of host cell death mechanisms. These hub and bottleneck genes represent possible targets for immunomodulation of host macrophages by virulent mycobacterial species that enable their survival within a hostile environment. Affymetrix GeneChipM-BM-. Bovine Genome Arrays were used to examine gene expression from a paired comparison of bovine monocyte-derived macrophages (MDM) after in vitro challenge with Mycobacterium bovis versus M. bovis BCG across a time series of 2 hr, 6 hr and 24 hr post-challenge.
Project description:Mycobacterium bovis is an intracellular pathogen that causes tuberculosis in cattle. Following infection, the pathogen resides and persists inside host macrophages by subverting host immune responses via a diverse range of mechanisms. Here, a high-density bovine microarray platform was used to examine the bovine monocyte-derived macrophage transcriptome response to M. bovis infection relative to infection with the attenuated vaccine strain, M. bovis Bacille Calmette–Guérin. Differentially expressed genes were identified (adjusted P-value ≤ 0.01) and interaction networks generated across an infection time course of 2, 6 and 24 h. The largest number of biological interactions was observed in the 24 h network, which exhibited small-worldscale-free network properties. The 24 h network featured a small number of key hub and bottleneck gene nodes, including IKBKE, MYC, NFKB1 and EGR1 that differentiated the macrophage response to virulent and attenuated M. bovis strains, possibly via the modulation of host cell death mechanisms. These hub and bottleneck genes represent possible targets for immunomodulation of host macrophages by virulent mycobacterial species that enable their survival within a hostile environment.
Project description:Geographical distinct virulent Babesia bovis strains have similar gene expression changes as they go through attenuation. Pair end RNA-sequencing reads on three biological replicate sample pairs of virulent parent and attenuated derivative Babesia bovis strain isolated in Argentina.
Project description:Infection of cattle with Mycobacterium bovis causes severe financial hardship in many countries, in addition to presenting a health risk for humans. As an intracellular pathogen, M. bovis, adapted to survive and thrive within the intramacrophage environment. However, little is known about expression patterns in the macrophage, particularly in the bovine host. In this study, DNA microarray analysis was used to detect genes expressed in Holstein bovine macrophages derived from peripheral blood mononuclear cells infected during four hours with two Argentinean strains of M. bovis, a virulent strain, 04-303 and an attenuated strain, 534. Genes encoding antrax toxin receptor, cell division and apoptosis regulator, ankyrin proteins that are found within the membrane associated cytoskeleton, protein of cell differentiation and regulators of endocytic traffic of membrane were more strongly expressed in infected macrophages. Blood from healthy Holstein bovines was taken in sterile conditions and peripheral blood mononuclear cells (PBMC) were separated from heparinized blood. PBMCs were used to prepare ten independent cultures which were incubated at 37C for one week in RPMI 1640 complete medium supplemented with 10% of autologous plasma. Four cultures were infected with viable cells of M. bovis virulent strain 04-303, four with avirulent strain 534 and two were left as uninfected controls. Four hours post infection, the cells were scraped, lysed. RNA was extracted, labeled and hybridized to ten Affymetrix Bovine Genome arrays.
Project description:The Toll-like receptor (TLR) and peptidoglycan recognition protein 1 (PGLYRP1) genes play key roles in the innate immune systems of mammals. While the TLRs recognize a variety of invading pathogens and induce innate immune responses, PGLYRP1 is directly microbicidal. We used custom allele-specific assays to genotype and validate 220 diallelic variants, including 54 nonsynonymous SNPs in 11 bovine innate immune genes (TLR1-TLR10, PGLYRP1) for 37 cattle breeds. Bayesian haplotype reconstructions and median joining networks revealed haplotype sharing between Bos taurus taurus and Bos taurus indicus breeds at every locus, and we were unable to differentiate between the specialized B. t. taurus beef and dairy breeds, despite an average polymorphism density of one locus per 219 bp. Ninety-nine tagSNPs and one tag insertion-deletion polymorphism were sufficient to predict 100% of the variation at all 11 innate immune loci in both subspecies and their hybrids, whereas 58 tagSNPs captured 100% of the variation at 172 loci in B. t. taurus. PolyPhen and SIFT analyses of nonsynonymous SNPs encoding amino acid replacements indicated that the majority of these substitutions were benign, but up to 31% were expected to potentially impact protein function. Several diversity-based tests provided support for strong purifying selection acting on TLR10 in B. t. taurus cattle. These results will broadly impact efforts related to bovine translational genomics.
Project description:Whole-transcriptome, high-throughput RNA sequence analysis of the bovine macrophage response to Mycobacterium bovis infection in vitro
Project description:Infection of cattle with Mycobacterium bovis causes severe financial hardship in many countries, in addition to presenting a health risk for humans. As an intracellular pathogen, M. bovis, adapted to survive and thrive within the intramacrophage environment. However, little is known about expression patterns in the macrophage, particularly in the bovine host. In this study, DNA microarray analysis was used to detect genes expressed in Holstein bovine macrophages derived from peripheral blood mononuclear cells infected during four hours with two Argentinean strains of M. bovis, a virulent strain, 04-303 and an attenuated strain, 534. Genes encoding antrax toxin receptor, cell division and apoptosis regulator, ankyrin proteins that are found within the membrane associated cytoskeleton, protein of cell differentiation and regulators of endocytic traffic of membrane were more strongly expressed in infected macrophages.
Project description:BackgroundWe present here the assembly of the bovine genome. The assembly method combines the BAC plus WGS local assembly used for the rat and sea urchin with the whole genome shotgun (WGS) only assembly used for many other animal genomes including the rhesus macaque.ResultsThe assembly process consisted of multiple phases: First, BACs were assembled with BAC generated sequence, then subsequently in combination with the individual overlapping WGS reads. Different assembly parameters were tested to separately optimize the performance for each BAC assembly of the BAC and WGS reads. In parallel, a second assembly was produced using only the WGS sequences and a global whole genome assembly method. The two assemblies were combined to create a more complete genome representation that retained the high quality BAC-based local assembly information, but with gaps between BACs filled in with the WGS-only assembly. Finally, the entire assembly was placed on chromosomes using the available map information.Over 90% of the assembly is now placed on chromosomes. The estimated genome size is 2.87 Gb which represents a high degree of completeness, with 95% of the available EST sequences found in assembled contigs. The quality of the assembly was evaluated by comparison to 73 finished BACs, where the draft assembly covers between 92.5 and 100% (average 98.5%) of the finished BACs. The assembly contigs and scaffolds align linearly to the finished BACs, suggesting that misassemblies are rare. Genotyping and genetic mapping of 17,482 SNPs revealed that more than 99.2% were correctly positioned within the Btau_4.0 assembly, confirming the accuracy of the assembly.ConclusionThe biological analysis of this bovine genome assembly is being published, and the sequence data is available to support future bovine research.
Project description:In this study we compared the transcriptome of bovine macrophages infected with two Mycobacterium bovis strains of variable virulence, co-cultured with autologous lymphocytes. We used the highly virulent M. bovis strain called Mb04-303 and the attenuated Mb534 strain. We first observed that only the infection of bovine macrophages with the virulent strain, Mb04-303, induced in peripheral bovine mononuclear cells a powerful innate immune response capable of controlling the intracellular mycobacterial replication.By RNAseq analysis we found that infections with Mb04-303 downregulated the KEAP1-NFE2L2 pathway that encodes a transcriptional factor involved in antioxidant genes and inflammasome activation and upregulated the type 1 interferon signalling pathway, compared to the infections with the attenuated Mb534 strain. T
Project description:Mycoplasma bovis (M.bovis) is a critical pathogen of bovines resulting in pneumonia, mastitis, arthritis, etc. To reveal its virulence related factors, a virulent M. bovis HB0801 and its derived vaccine strain P150 were infected calve and the transcriptome profiles of PBMCs were compared by using of the microarray at 7 days after infection. The data postulated the pathogenic mechanism of wild strain and immune mechanism of the attenuated strain to provide clues for the further research of the interaction between M. bovis and its host.