Transcription profiling of bovine peripheral blood mononuclear cells stimulated with Mycoplasma bovis against untreated controls
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ABSTRACT: Mycoplasma species are highly contagious pathogens, and Mycoplasma infectious disease are a serious issue for the dairy industry. The bovine peripheral blood mononuclear cells (PBMCs) play an important role for the eradication of pathogens which cause mycoplasmal infection, however the effects of M. bovis for immune response of PBMCs in vitro have not been fully clarified.We examined the transcription profiling of bovine PBMCs on the stimulation with M. bovis for 6h (3 stimuli, 3 control).
Project description:Mycoplasma species are highly contagious pathogens, and Mycoplasma infectious disease are a serious issue for the dairy industry. The bovine peripheral blood mononuclear cells play an important role for mycoplasma mastitis, however, the effects of M. bovis for immune response of peripheral blood mononuclear cells have not been fully clarified.We examined the transcription profiling of bovine peripheral blood mononuclear cells in intramammary infusion of M. bovis at day 7.
Project description:Mycoplasma species are highly contagious pathogens, and Mycoplasma infectious disease are a serious issue for the dairy industry. The bovine neutrophils play an important role for the eradication of pathogens which cause mycoplasmal infection, however the effects of M. bovis for immune response of neutrophils have not been fully clarified. We examined the transcription profiling of bovine neutrophils on the stimulation with M. bovis for 3h (3 stimuli, 3 control).
Project description:Mycoplasma species are highly contagious pathogens, and intramammary Mycoplasma infection is a serious issue for the dairy industry. The bovine mammary epithelial cells (bMEC) play an important role for the eradication of pathogens which cause intramammary infection, however the effects of M. bovis for immune response of bMEC have not been fully clarified. We examined the transcription profiling of bMEC on the stimulation with M. bovis for 6h (3 stimuli, 3 control).
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:Mycoplasma bovis is one of the major causative pathogens of the bovine respiratory complex disease that is characterized by enzootic pneumonia, mastitis, pleuritis and polyarthritis. M. bovis enters and colonizes the bovine respiratory epithelia through inhalation of aerosol from contaminated air. The nature of the interaction between M. bovis and bovine innate immune system is not well understood. We hypothesized that M. bovis invades blood monocytes and regulates cellular function to support its persistence and systemic dissemination. We used bovine-specific peptide kinome arrays to identify cellular signaling pathways that could be relevant to M. bovis-monocyte interaction in vitro. We validated these pathways using functional, protein and gene expression arrays. Here we show that infection of blood monocytes with M. bovis delays spontaneous or TNF-α/staurosporine-driven apoptosis, activates NF-κβ p65 subunit and inhibits caspase-9 activity. We also report that M. bovis infected bovine monocytes do not produce IFN-γ and TNF-α, although production of IL-10 is elevated. Our findings suggest that M. bovis takes over the cellular machinery of bovine monocytes to prolong bacterial survival and to possibly facilitate subsequent systemic distribution.
Project description:Micro RNA profiling was performed on in vitro PPDB and nil stimulated bovine PBMCs isolated from BCG vaccinated and unvaccinated cattle before and after challenge with M. bovis.
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:Background: Mycobacterium bovis, the causative agent of bovine tuberculosis, is a major cause of mortality in global cattle populations. Macrophages are among the first cells types to encounter M. bovis following exposure and the response elicited by these cells is pivotal in determining the outcome of infection. Here, a functional genomics approach was undertaken to investigate global gene expression profiles in bovine monocyte-derived macrophages (MDM) purified from seven age-matched non-related females, in response to in vitro challenge with M. bovis (multiplicity of infection 2:1). Total cellular RNA was extracted from non-challenged control and M. bovis-challenged MDM for all animals at intervals of 2 hours, 6 hours and 24 hours post-challenge and prepared for global gene expression analysis using the Affymetrix® GeneChip® Bovine Genome Array. Results: Comparison of M. bovis-challenged MDM gene expression profiles with the non-challenged MDM controls at each time point identified 3,529 differentially expressed genes after 2 hours post-challenge, with 5,211 and 6,150 differentially expressed genes detected at the 6 hour and 24 hour time points, respectively (adjusted P-value threshold ≤ 0.05). Notably, the number of downregulated genes exceeded the number of upregulated genes in the M. bovis-challenged MDM across all time points; however, the fold-change in expression for the upregulated genes was markedly larger than that for the downregulated genes. Systems analysis revealed enrichment for genes involved in: (1) the inflammatory response; (2) cell signalling pathways, including Toll-like receptors and intracellular pathogen recognition receptors (PRRs)-receptors; and (3) apoptosis. Conclusions: The increased number of downregulated genes is consistent with previous studies showing that M. bovis infection is associated with the repression of host gene expression. The results also support important roles for MYD88-independent signalling and intracellular PRRs in mediating the host response to M. bovis, which to our knowledge have not been reported previously. Affymetrix GeneChip® Bovine Genome Arrays were used to examine gene expression of bovine monocyte-derived macrophages (MDM) after in vitro challenge with Mycobacterium bovis across a time series of 2 hr, 6 hr and 24 hr post-challenge. A 0 hr control treatment was also generated and seven different age-matched female Holstein-Friesian cattle were used for each time-point/treatment combination for a total of 49 microarrays.
Project description:The immune response associated with mastitis caused by Mycoplasma bovis is a very complicated biological process in several type of cells, including immune cells, mammary epithelial cells and, endothelial cells. Thus, revealing of the microRNAs in the Mycoplasma bovis infected mammary gland tissues is particularly important for the immune response mechanism to Mycoplasma bovis. Firstly, mammary gland tissue samples were collected from Holstein cows and screened for Mycoplasma bovis. Then, total RNA was isolated from mycoplasma bovis infected tissues and RNA sequencing was performed. After bioinformatics analysis, GO and KEGG analysis of target genes of identified microRNAs were conducted. Our results revaled that 24 of the known microRNAs were expressed differently and 13 of the novel microRNAs were expressed differently in Mycoplasma bovis positive tissues. The target genes of these microRNAs were found to be associated with especially inflammation pathways. In conclusion, this study demonstrated that identified miRNAs may be involved in the signaling pathways during mastitis case caused by Mycoplasma bovis.
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.