Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Transcription profiling of mouse splenocytes response to infection with wild type or virB mutant Brucella strains

ABSTRACT: The virB operon, encoding a Type IV secretion system (T4SS), is essential for intracellular survival and persistent infection of Brucella spp. To better understand the role of the T4SS in evading host defense mechanisms and establishing chronic infection, we compared transcriptional profiles of the host response to infection with wild type Brucella strains and strains that fail to express the virB genes. Analysis of host gene expression profiles three days after inoculation with wild type Brucella strains revealed an inflammatory response dominated by interferon-induced genes. This analysis found that not only the type II but also type I interferon pathway was elicited by Brucella infection. Real time RT-PCR showed that a group of genes from these pathways was induced by day 3 post-infection and declined to baseline levels by day 7. In contrast, neither of the two virB mutant strains elicited expression of interferon-induced genes, demonstrating that the T4SS was required to trigger an inflammatory response early during infection. Experiment Overall Design: We performed microarray-based expression analyses of splenocytes from mice infected with two virulent strains (B. abortus 2308 and B. melitensis 16M), and two different B. abortus virB mutants, whose virB operon was either disrupted (BA41) or completely deleted (ADH4.2), to better understand the contribution of the T4SS in establishing infection of the reticuloendothelial system. 3 days after infection of mice, spleens were excised for RNA extraction. For each bacterial strain, RNA from 5 mice was pooled and reverse transcribed for hybridization to an array. Each experiment was performed in duplicate.

ORGANISM(S): Mus musculus

SUBMITTER: Renee Tsolis

PROVIDER: E-GEOD-6810 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Brucella requires a functional Type IV secretion system to elicit innate immune responses in mice.

Roux Christelle M CM Rolán Hortensia G HG Santos Renato L RL Beremand Phillip D PD Thomas Terry L TL Adams L Garry LG Tsolis Renée M RM

Cellular microbiology 20070417 7

The virB operon, encoding a Type IV secretion system (T4SS), is essential for intracellular survival and persistent infection by Brucella spp. To better understand the role of the T4SS in evading host defence mechanisms and establishing chronic infection, we compared transcriptional profiles of the host response to infection with wild-type and virB mutant Brucella strains. Analysis of gene expression profiles in murine splenocytes 3 days after inoculation with wild-type Brucella strains revealed ...[more]

PMID: 17441987

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Project description:Brucellosis is still a widespread zoonotic disease. Very little is known about the interaction between B. abortus and trophoblastic cells, which is essential for better understanding the pathogenesis of the Brucella-induced placentitis and abortion, a key event for transmission of the disease. The goal of this study was to evaluate the profile of gene expression by bovine trophoblastic cells during infection with B abortus. Explants of chorioallantoic membranes were inoculated with B. abortus strain 2308. Microarray analysis was performed at 4 h after infection, and expression of cytokines and chemokines by trophoblastic cells was assessed by real time RT-PCR at 6 and 12 h after inoculation. In addition, cytokine and chemokine expression was evaluated in placentomes from experimentally infected cows. Expression of pro-inflammatory genes by trophoblastic cells was suppressed at 4 h after inoculation, whereas a significant up-regulation of CXC chemokines, namely CXCL6 (GCP-2) and CXCL8 (IL-8), was observed at 12, but not at 6 h after inoculation. Placentomes of experimentally infected cows had a similar profile of chemokine expression, with upregulation of CXCL6 and CXCL8. Our data indicate that B. abortus modulates the innate immune response by trophoblastic cells, suppressing expression of pro-inflammatory mediators during the early stages of infection that is followed by a delayed and mild expression of pro-inflammatory chemokines, which is similar to the profile of chemokine expression in the placentomes of experimentally infected cows. This trophoblastic response is likely to contribute to the pathogenesis of B. abortus-induced placentitis. Keywords: trophoblast response to Brucella Total RNA used for microarray analysis was obtained from 6 placentas with 12 infected and 12 control explants from each placenta. At 4 h after inoculation, total RNA was isolated from the trophoblastic surface of the explants. Three RNA pools from two placentas each were generated prior to cDNA synthesis.

Dataset Information

Transcription profiling of mouse splenocytes response to infection with wild type or virB mutant Brucella strains

Publications

Brucella requires a functional Type IV secretion system to elicit innate immune responses in mice.

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