Transcriptome analysis of Streptococcus suis serotype 2 during infection in a murine air pouch model
Ontology highlight
ABSTRACT:
ORGANISM(S): Streptococcus suis
SUBMITTER: Chengkun Zheng
PROVIDER: E-GEOD-68032 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
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Project description:During infection in the air pouches, S. suis encounters changing environments and host factors. It is important for S. suis to modulate the gene expression in response to stimulants. Using DNA microarray analysis, we compared the transcriptome of S. suis obtained from the air pouches with that from working cultures.
2015-04-25 | GSE68032 | GEO
Project description:Objective. To identify novel monosodium urate (MSU) crystal-induced mRNAs by transcript profiling of isolated murine air pouch membranes. Methods. Nine hours after injecting crystals into air pouches, membranes were meticulously dissected away from the adjacent soft tissues. mRNA expression differences between inflamed and control membranes were determined by oligonucleotide microarray analysis. Induction of selected mRNAs was validated by real-time relative quantitative reverse transcriptase PCR (qPCR) in pouch membranes and murine peritoneal macrophages. Results. Eleven of the 12 most highly upregulated mRNAs related to innate immunity and inflammation. They included mRNAs encoding histidine decarboxylase (the enzyme that synthesizes histamine), interleukin (IL)-6, the cell surface receptors PUMA-g and TREM-1, and the polypeptides Irg1 and PROK-2. MSU crystals induced dramatic rises in these mRNAs in the pouch membrane within 3-8 hours after the surge in pro-inflammatory cytokine (IL-6, IL-1beta and TNFalpha) and immediate early gene (Egr-1) transcription, which occurred 1h after crystal injection. MSU crystals induced these mRNAs in cultured macrophages with similar kinetics but lower fold changes. In keeping with their downregulation by MSU crystals according to the microarrays, qPCR confirmed that TREM-2 and granzyme D mRNAs decreased 79% and 94%, respectively, in MSU crystal inflamed membranes. Conclusions. This analysis disclosed several genes previously not implicated in MSU crystal inflammation. Their rise after the early surge in cytokine mRNAs suggests that they may, for instance, amplify or perpetuate inflammation. Transcript profiling of the isolated air pouch membrane promises to be a powerful tool to identify genes acting at different stages of inflammation. Experiment Overall Design: Mouse strain: female Balb/C, 6-8 weeks old Experiment Overall Design: Number of mice= 6. Experiment Overall Design: Air pouches raised on back for 7 days by injecting 2ml of sterile air. Experiment Overall Design: On day 7, injection of 1ml PBS or 2mg MSU crystals in 1ml PBS (n=3 each) Experiment Overall Design: 9 h after injection, dissection of the air pouch membranes from the adjacent soft tissues, RNA extraction with RNeasy columns. Pooling of RNA from the 3 control pouches or the 3 MSU crystal pouches. Processing and labeling of 5ug aliquots in the U Penn microarray core according to standard Affy protocols. Hybridization to Affy Mo430_2 oligonucleotide microarrays.
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Project description:Mast cells (MCs) were implicated in tissue remodelling in hamster models of heart inflammatory diseases. Here we asked whether MCs promote inflammatory neovascularization in hamster cheek pouch (HCP) parasitized by Trypanosoma cruzi, the protozoan that causes Chagas disease. Intravital microscopy (IVM) performed 3 days after inoculation of Dm28c trypomastigote (GFP-TCTs) showed a subtle infiltration of rhodamine-labeled leukocytes. Interestingly, the baseline levels of FITC-dextran leakage (3 dpi) were consistently increased, raising the possibility that intracellular amastigotes may coopt the low-grade inflammation to facilitate the delivery of plasma-borne nutrients during the period in which host cells must metabolically sustain parasite division. Although proangiogenic indexes were only mildly stimulated at 3 dpi, we found a positive correlation with transcription of proinflammatory cytokines (pro-IL- and IFN-). Having established 3 dpi as a timepoint in which vascular remodeling was already measurable, we next compared the proteomic profiles of parasitized HCP with those of internal controls (PBS injected 3 days earlier in the contralateral pouch) we found 87 proteins upregulated, and 17 were downregulated in the parasitized HCP. Minor changes in protein expression were observed in response to PBS injection (2 upregulated and 12 downregulated proteins. Congruent with the increased density of MCs observed in parasitized HCP (histological analysis), the MC protease 1 (chymase) stood out as the most abundant sequence. Interestingly, toluidine blue staining appointed clusters of MCs degranulating at 3 dpi. Noteworthy, two plasma proteins (complement C3 and serum albumin) were also identified as major proteins, further suggesting that leaky capillaries irrigated the HCP at early stages (3 dpi) of infection . Detection of cytokines or classical proangiogenic factors was beyond the sensitivity of our methods.
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Project description:Streptococcus suis is an important emerging worldwide pig pathogen and zoonotic agent with rapid evolution of virulence and drug resistance. Licochalcone A, used in traditional Chinese medicine, exhibits antimicrobial, antioxidant and anti-inflammatory activities. Herein, a whole-genome DNA microarray was used to investigate the global transcriptional regulation of Streptococcus suis 05ZYH33 treated by subinhibitory concentration of licochalcone A. 132 genes were differentially regulated upon liochalcone A treatment, including 78 genes up-regulated and 54 genes down-regulated which included many central biological functions such as metabolism, transcription and translation. We tried to investigate the antimicrobial mechanism of licochalcone A in the aspect of bacterial cell cycle control. Our analysis indicated that licochalcone A might inhibit the growth of S. suis by controlling the replication initiation and cell division through amino acid metabolism. A cDNA microarray imprinted with 2156 genes representing about 98% of Streptococcus suis serotype 2 genome was used for transcriptome analysis. For two-sample (reference vs. test) microarray hybridization, four independent bacterial cultures from each condition were prepared as biological replicates for RNA isolation. Four dual-fluorescence-labeled cDNA probes were prepared to hybridize with four slides, respectively. Pairwise comparisons were made using dye swaps to avoid labeling bias. A ratio of mRNA levels (test/reference) was calculated for each gene. Significant changes of gene expression were identified with the SAM software. After the SAM analysis, only genes with at least 2-fold changes in expression were collected for further analysis.
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Project description:We investigated for the first time the in vitro response of PRRSV-infected porcine DCs and monocytes to S. suis. We first assessed the effect of PRRSV infection on the phagocytosis and intracellular survival of S. suis by these two cell populations. We then used a genomic approach to compare the gene expression profiles of both type of cells infected with S. suis, with or without a previous infection with PRRSV. Total RNA obtained from porcine monocytes and dendritic cells infected with S. suis, PRRSV, or S. suis & PRRSV. Four replicates in all groups.
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