Transcriptional profiling of spleens from mice challenged with Brucella melitensis with or without prior vaccination.
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ABSTRACT:
ORGANISM(S): Mus musculus
PROVIDER: GSE254248 | GEO | 2024/01/30
REPOSITORIES: GEO
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Project description:In this study we attempt to elucidate some of the pathways involved in the immune response to vaccination and subsequent disease challenge using a transcriptomic approach. We exposed, via intra-peritoneal injection, three months old Asian seabass (Lates calcarifer) to a Streptococcus iniae vaccine. A control group was also set up where no vaccine was injected. Spleen and head kidney samples were collected at one and seven days post vaccination for transcriptomic analysis. At this point, there are four groups per organ: Day 1 vaccinated, Day 1 control, Day 7 vaccinated and Day 7 control. Subsequently, a pathogen challenge was carried out three weeks later and spleen and head kidneys were sampled at 25-29 hours post challenge for transcriptomic analysis. For control, mock challenged was carried out. At this point, there are four groups per organ: unvaccinated challenged, unvaccinated mock challenged, vaccinated challenged and vaccinated mock challenged. Total 57 samples. Spleen samples: At Day 1 and Day 7 post vaccination, 4 spleens were analyzed for each of the D1 control and D1 vaccinated groups and 3 spleens were analyzed for each of the D7 control and D7 vaccinated groups (total = 14 samples). At post pathogen challenge, 4 spleens were analyzed for each of these three groups - unvaccinated mock challenged, vaccinated challenged and vaccinated mock challenged and 3 spleens for the unvaccinated challenged group (total = 15 samples). Head Kidney samples: At Day 1 and Day 7 post vaccination, 3 head kidneys were analyzed for the control and vaccinated groups for both time points (total = 12 samples). At post pathogen challenge, 4 head kidneys were analyzed for each of the groups: unvaccinated challenged, unvaccinated mock challenged, vaccinated challenged and vaccinated mock challenged (total = 16 samples).
2014-10-01 | E-GEOD-51839 | biostudies-arrayexpress
Project description:Brucellosis is a zoonosis of the Mediterranean and Middle-East regions linked to important economic losses and reduced animal welfare. To date, no effective diagnostic and/or prophylactic measures are available for the control of brucellosis, due to the residual virulence of the bacterial strain administered for vaccinal purposes and the difficulties in distinguishing vaccinated from infected animals. To overcome these issues, studies are desired to elucidate the bacterial biology and the pathogenetic mechanisms of both the vaccinal strain and the pathogenic strains. Here, we employed a label-free shotgun proteomics to investigate the protein repertoire of the vaccinal strain B. melitensis Rev.1 and compare it with the proteome of the Brucella melitensis 16 M, one of the most common field strains isolated from ruminants. Comparative proteomics profiling underlines common and diverging traits between the two strains, providing suggestions on the potential biochemical routes responsible of the residual virulence of the vaccinal strain; whilst the diverging traits are suggestive biochemical signatures to be further investigated to provide an optimized diagnostics capable of discriminating the vaccinated from infected animals.
2021-05-20 | PXD022472 | Pride
Project description:Brucella ovis causes an important disease characterized by decreased fertility in rams, sporadic abortions in ewes and increased lamb mortality. The live attenuated B. melitensis Rev.1 vaccine is considered the best vaccine available against this infection. However, this vaccine shows variable protective efficacy ranging from 40% to 100%.The objective of this study was to identify possible correlates of protective response to B. ovis infection through the characterization by microarray hybridization and real-time RT-PCR of inflammatory and immune response genes upregulated in rams previously immunized with the B. melitensis Rev 1 vaccine strain and experimentally challenged with B. ovis. Gene expression profiles were compared before and after challenge with B. ovis between rams protected and those vaccinated but found infected after challenge. The genes upregulated in vaccinated and protected rams provide possible correlates of protective response to B. ovis infection in rams immunized with the B. melitensis Rev 1 vaccine. Gene expression profiles were compared before and after vaccination and challenge with B. ovis between rams protected with the B. melitensis Rev 1 vaccine strain and those vaccinated but infected after challenge. Total RNA was isolated from buffy coat samples before vaccination (T0), after vaccination and before challenge (T1) and 60 days post challenge (T2) using TriReagent (Sigma, St. Louis, MO, USA)
2010-06-25 | E-GEOD-13202 | biostudies-arrayexpress
Project description:Brucella ovis causes an important disease characterized by decreased fertility in rams, sporadic abortions in ewes and increased lamb mortality. The live attenuated B. melitensis Rev.1 vaccine is considered the best vaccine available against this infection. However, this vaccine shows variable protective efficacy ranging from 40% to 100%.The objective of this study was to identify possible correlates of protective response to B. ovis infection through the characterization by microarray hybridization and real-time RT-PCR of inflammatory and immune response genes upregulated in rams previously immunized with the B. melitensis Rev 1 vaccine strain and experimentally challenged with B. ovis. Gene expression profiles were compared before and after challenge with B. ovis between rams protected and those vaccinated but found infected after challenge. The genes upregulated in vaccinated and protected rams provide possible correlates of protective response to B. ovis infection in rams immunized with the B. melitensis Rev 1 vaccine.
2008-10-17 | GSE13202 | GEO
Project description:Brucella spp. is an intracellular pathogen in vivo. The intracellular B. melitensis transcriptome was determined by initially enriched and then amplified B. melitensis RNA from total RNA of B. melitensis-infected HeLa cells. Analysis of microarray results identified 161 and 115 genes differentially expressed at 4 and 12 h p.i., respectively. Most of the genes (78%) differentially expressed were down-regulated at the earliest time point, but up-regulated (75%) at 12 h p.i. The analysis of the results indicates that Brucella undergo an adaptation period during the first 4 h p.i. that is overcome by 12 h p.i., permitting Brucella to replicate intracellularly. Specific genes and biological processes identified in this study will further help elucidate how Brucella act during the early infectious process to their eventual benefit and to the detriment of the naM-CM-/ve host. Keywords: Time course study of intracellular B. melitensis gene expression Gene expression of the intracellular Brucella melitensis was determined at 4 and 12 h p.i. We generated the following samples: A) B. melitensis total RNA enriched and amplified from total RNA of B. melitensis-infected HeLa cells at 4 h p.i.; B) Total RNA isolated from B. melitensis-infected HeLa cells at 4 h p.i.; C) B. melitensis total RNA enriched and amplified from total RNA of B. melitensis-infected HeLa cells at 12 h p.i.; D) Total RNA isolated from B. melitensis-infected HeLa cells at 12 h p.i. B. melitensis total RNA was initially enriched and then amplified from total RNA of B. melitensis-infected HeLa cells at 4 and 12 h p.i. in quadruplicate, indirectly labeled and co-hybridized against B. melitensis gDNA to a custom 3.2K B. melitensis oligo-array (n = 8). As there was a possibility that some HeLa transcripts cross-hybridize with probes on B. melitensis microarrays, the original total RNA from B. melitensis-infected HeLa cells were also co-hybridized against B. melitensis gDNA to B. melitensis oligo-arrays (n = 8), and any oligospots with signals were considered non-specific and eliminated from all analysis to avoid false positive gene detection. The intracellular B. melitensis gene expression was compared to the gene expression of the inoculum (n = 2). Every Brucella melitensis open reading frame was printed in triplicate on each microarray, thereby providing three technical replicates for each biological replicates. Each replicate was normalized against labeled Brucella melitensis genomic DNA.
2012-12-27 | E-GEOD-14704 | biostudies-arrayexpress
Project description:Brucella spp. is an intracellular pathogen in vivo. The intracellular B. melitensis transcriptome was determined by initially enriched and then amplified B. melitensis RNA from total RNA of B. melitensis-infected HeLa cells. Analysis of microarray results identified 161 and 115 genes differentially expressed at 4 and 12 h p.i., respectively. Most of the genes (78%) differentially expressed were down-regulated at the earliest time point, but up-regulated (75%) at 12 h p.i. The analysis of the results indicates that Brucella undergo an adaptation period during the first 4 h p.i. that is overcome by 12 h p.i., permitting Brucella to replicate intracellularly. Specific genes and biological processes identified in this study will further help elucidate how Brucella act during the early infectious process to their eventual benefit and to the detriment of the naïve host. Keywords: Time course study of intracellular B. melitensis gene expression
2012-12-27 | GSE14704 | GEO
Project description:Investigation of whole genome gene expression level changes in a Brucella melitensis delta prlr mutant compared to the wild type strain. The mutants analyzed in this study are further described in A. Mirabella, R-M Yanez, R.M. Delrue, S. Uzureau, M.S. Zygmunt, A. Cloeckaert, X. De Bolle, J.J. Letesson (2012). The two component system PrlS/PrlR of Brucella melitensis is required for persistence in mice and appears to respond to ionic strength. Microbiology
2012-12-31 | GSE34633 | GEO
Project description:In this study we report that B. melitensis at the late logarithmic phase of growth are more invasive for HeLa cells than at mid logarithmic or stationary growth phases. Microarray analysis of B. melitensis gene expression identified 414 up- and 40 down-regulated genes in late-log growth phase compared to the stationary growth phase. The vast majority of the up-regulated genes in late-log cultures were those associated with DNA replication, transcription and translation, intermediate metabolism, energy production and conversion, membrane transport and cell envelope, biogenesis and outer membrane, while the down-regulated genes were distributed among several functional categories. This first Brucella global gene expression study provides novel information on growth phase-specific gene regulation important not only for understanding Brucella physiology but also the initial molecular interactions between Brucella and its host. Keywords: Comparison bacterial growth phase normalized to genomic DNA There are two kind of samples consisting of RNA isolated from Brucella melitensis grown logarithmically or at stationary phase. There are four biological replicates of each sample. Every Brucella melitensis open reading frame was printed in triplicate on each microarray, thereby providing three technical replicates for each biological replicates. Each replicate was normalized against labeled Brucella melitensis genomic DNA.
2010-05-19 | E-GEOD-11192 | biostudies-arrayexpress
Project description:Brucella type strains (B. abortus 544 and B. melitensis 16 M), field-isolates (B. abortus T and B. melitensis C) were independently cultivated six times on Tryptic Soy Broth at 37 °C in the presence of 5% CO2 until mid-logarithmic phase. The cells were harvested by brief centrifugation and the bacteria was inactivated using recommended ethanol precipitation procedure. Following which the cells were lysed, proteins quantified and the differentially expressed proteins among B. abortus and B. melitensis were identified using liquid chromatography-mass spectrometry (LC/MS) based label free quantitative proteomics analysis.
2020-06-11 | PXD006348 | Pride
Project description:Investigation of whole genome gene expression level changes in a Brucella melitensis delta prlr mutant compared to the wild type strain. The mutants analyzed in this study are further described in A. Mirabella, R-M Yanez, R.M. Delrue, S. Uzureau, M.S. Zygmunt, A. Cloeckaert, X. De Bolle, J.J. Letesson (2012). The two component system PrlS/PrlR of Brucella melitensis is required for persistence in mice and appears to respond to ionic strength. Microbiology A six chip study using total RNA recovered from three separate wild-type cultures of Brucella melitensis 16M and three separate cultures of a prlR mutant strain. Each chip measures the expression level of 3,198 genes from Brucella melitensis 16M with nineteen 60 mer probe pairs (PM/MM) per gene, with three-fold technical redundancy.
2012-12-31 | E-GEOD-34633 | biostudies-arrayexpress