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ABSTRACT: Northern Ireland Brucella abortus

PROVIDER: PRJNA576561 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR10249500_1.fastq.gz	Fastqsanger.gz
	SRR10249500_2.fastq.gz	Fastqsanger.gz
	SRR10249501_1.fastq.gz	Fastqsanger.gz
	SRR10249501_2.fastq.gz	Fastqsanger.gz
	SRR10249502_1.fastq.gz	Fastqsanger.gz

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Gene expression profiling of Brucella abortus-infected mouse macrophages

Project description:Gene expression analysis of wild-type and STING knock-out mouse bone marrow-derived macrophages (mBMDM) infected with Brucella abortus or transfected with Brucella abortus DNA. Genes whose expression are affected by Brucella abortus in a STING-dependent manner will be identified and signaling pathways regulated by STING will be elucidated.

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Proteomic Analysis of Stationary Growth Stage and Nutritional Deficiency Adaptation Reveals Brucella Abortus Virulence Mechanisms

Project description:Brucellosis, an important bacterial zoonosis caused by Brucella species, has drawn increased attention around the world. As an intracellular pathogen, the ability of Brucella to deal with stress within the host cell is closely related to its virulence. The survival pressure on Brucella within a phagosome is considered similar to that during the stationary phase. Here, label-free proteomics approach was used to study the adaptive response of Brucella abortus (B. abortus) in the stationary stage. 182 down-regulated and 140 up-regulated proteins were found in the stationary-phase B. abortus. B. abortus adapted to adverse environmental changes by regulating virulence, reproduction, transcription, translation, stress response, and energy production. In addition, both logarithmic and stationary-phase B. abortus were treated with short-term starvation. The logarithmic-phase B. abortus restricted cell reproduction and energy utilization in response to nutritional stress. Additionally, the expression levels of some virulence-related proteins were identified as being significantly regulated during the transition from logarithmic to stationary phase or under starvation treatment, such as Type IV secretion system protein (T4SS), VjbR, and integration host factor (IHF). Altogether, we outlined adaptive mechanisms that B. abortus could employ during the growth and compared the differences between logarithmic and stationary-phase B. abortus in response to starvation.

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Project description:We focused on whether transposon mutagenesis in Brucella abortus could induce difference in the trascriptional responses of RAW 264.7 cell infection model compared to the wild strain infected RAW 264.7 cells. The function of genes in Brucella abortus was analyzed through the identified differences in gene expression between RAW 264.7 cell infected with wild and mutant strains. We analyzed altered transcription in RAW 264.7 cells at 0, 6, 12, and 24 h following the infection with 10 MOI of Brucella abortus wild and mutant strains.

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Project description:Identification of host responses at the gene transcription level provides a molecular profile of the events that occur following infection. Brucella abortus is a facultative intracellular pathogen of macrophages that induces chronic infection in humans and domestic animals. Using microarray technology, the response of macrophages 4 hours following B. abortus infection was analyzed to identify early intracellular infection events that occur in macrophages. Of the more than 6,000 genes, we identified over 140 genes that were reproducibly differentially transcribed. First, an increase in the transcription of a number of pro-inflammatory cytokines and chemokines, such as TNF-α, IL-1β, IL-1α, and members of the SCY family of proteins, was evident that may constitute a general host recruitment of antibacterial defenses. Alternatively, Brucella may subvert newly arriving macrophages for additional intracellular infection. Second, transcription of receptors and cytokines associated with antigen presentation, e.g., MHC class II and IL-12p40, were not evident at this 4 hour period of infection. Third, Brucella inhibited transcription of various host genes involved in apoptosis, cell cycling, and intracellular vesicular trafficking. Identification of macrophage genes whose transcription was inhibited suggests that Brucella utilizes specific mechanisms to target certain cell pathways. In conclusion, these data suggest that B. abortus can alter macrophage pathways to recruit additional macrophages for future infection while simultaneously inhibiting apoptosis and innate immune mechanisms within the macrophage permitting intracellular survival of the bacterium. These results provide insights into the pathogenic strategies used by Brucella to survive long-term within a hostile environment. Keywords: Macrophage, intracellular pathogen, Brucella abortus, inflammatory immune response

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