Unknown,Transcriptomics,Genomics,Proteomics

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Gene microarray analysis of mouse-brain endothelial cells exposure to Pertussis Toxin and B-oligomer

ABSTRACT: Pertussis Toxin (PTX) regulated inflammatory and angiogenesis-related transcripts in endothelial cells from mouse brain(MBEC). Of ~14,000 mouse genes tracked, 34 showed altered expression in response to PTX, and most of these changes were dependent on the ADP-ribosyltransferase activity of the S1 subunit of the active holotoxin. More than one third of these genes have roles in inflammation and angiogenesis. Keywords: response to treatment MBEC(passage1) cultured on collagen-coated glass chamber slides were treated with PTX and B-oligomer at 3ng/ml, or PBS as control. Six hours later, total RNA were isolated from the cells. Gene microarray were performed to identify early candidate genes that are affected by PTX. The experiment was repeated twice independently.

ORGANISM(S): Mus musculus

SUBMITTER: Jeffrey Bond

PROVIDER: E-GEOD-11288 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Pertussis is a highly contagious, acute respiratory disease in humans caused by the Gram-negative pathogen Bordetella pertussis. Pertussis has resurged in the face of intensive vaccination and this has coincided with the emergence of strains carrying a particular allele for the pertussis toxin promoter, ptxP3, which is associated with higher levels of pertussis toxin (Ptx) production. Within 10 to 20 years, ptxP3 strains have nearly completely replaced the previously dominant ptxP1 strains resulting in a worldwide selective sweep. In order to identify B. pertussis genes associated with the selective sweep, we compared the expression of genes in ptxP1 and ptxP3 strains that are under control of the Bordetella master virulence regulatory locus (bvgASR). The BvgAS proteins comprise a two component sensory transduction system which is regulated by temperature, nicotinic acid and sulfate. By increasing the sulfate concentration, it is possible to change the phase of B. pertussis from virulent to avirulent. Until recently, the only distinctive phenotype of ptxP3 strains was a higher Ptx production. Here we identify additional phenotypic differences between ptxP1 and ptxP3 strains which may have contributed to its global spread by comparing global transcriptional responses under sulfate-modulating conditions. We show that ptxP3 strains are less sensitive to sulfate-mediated gene suppression, resulting in an increased production of the vaccine antigens pertactin (Prn) and Ptx and a number of other virulence genes, including a type III secretion toxin, Vag8, a protein involved in complement resistance, and lpxE involved in lipid A modification. Furthermore, enhanced expression of the vaccine antigens Ptx and Prn by ptxP3 strains was confirmed at the protein level. Identification of genes differentially expressed between ptxP1 and ptxP3 strains may elucidate how B. pertussis has adapted to vaccination and allow the improvement of pertussis vaccines by identifying novel vaccine candidates.

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Gene microarray analysis of mouse-brain endothelial cells exposure to Pertussis Toxin and B-oligomer

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