Unknown,Transcriptomics,Genomics,Proteomics

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Transcriptional profile of bovine chorioallantoic membrane explants in response to wild type, M-NM-^TvirB2 or M-NM-^TbtpB Brucella abortus

ABSTRACT: In this study the transcription profile of bovine trophoblastic cells infected with Brucella samples was evaluated. Chorioallantoic membrane (CAM) explants inoculated with wild type (strain 2308), and to compare with the transcription profile of trophoblastic cells infected with mutant strains lacking virB2 or btpB.M-NM-^TvirB2 or M-NM-^TbtpB by microarray analysis at 4 hours post infection. Genes with significant variation in levels of transcripts (fold change > 2 and P < 0.05) were functionally classified, and transcripts related to defense and inflammation were assessed by quantitative real time RT-PCR. Chorioallantoic membrane explant culture (CAM) were obtained from 7 intact pregnant bovine uteruses at the final third of gestation. Three-condition experiment, wild type B. abortus 2308, DvirB2 or DbtpB in 4 control replicates and Microarray analysis.

ORGANISM(S): Bos taurus

SUBMITTER: ALEX CARVALHO

PROVIDER: E-GEOD-58216 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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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.

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Transcriptional profile of bovine chorioallantoic membrane explants in response to wild type, M-NM-^TvirB2 or M-NM-^TbtpB Brucella abortus

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