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Gene expression and synergistic interactions induced by CpG ODN and/or poly(I:C)

ABSTRACT: Bioinformatic analysis of microarray data was used to identify the regulatory patterns underlying changes in gene expression induced when RAW 264.7 macrophages were stimulated via TLR9 by CpG oligonucleotides (ODN) and/or via TLR3 by poly (I:C). While the genes activated by each ligand mediated similar functions, poly (I:C) elicited a larger and more diverse change in gene expression. Co-stimulation with both ligands accelerated gene expression and synergistically activated genes primarily associated with immune function. This is the first work to compare global changes in gene regulation triggered by distinct TLR pathways and clarify their impact on gene expression.

ORGANISM(S): Mus musculus

PROVIDER: GSE15416 | GEO | 2009/07/21

SECONDARY ACCESSION(S): PRJNA115821

REPOSITORIES: GEO

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Gene expression and synergistic interactions induced by CpG ODN and/or poly(I:C)

Project description:Bioinformatic analysis of microarray data was used to identify the regulatory patterns underlying changes in gene expression induced when RAW 264.7 macrophages were stimulated via TLR9 by CpG oligonucleotides (ODN) and/or via TLR3 by poly (I:C). While the genes activated by each ligand mediated similar functions, poly (I:C) elicited a larger and more diverse change in gene expression. Co-stimulation with both ligands accelerated gene expression and synergistically activated genes primarily associated with immune function. This is the first work to compare global changes in gene regulation triggered by distinct TLR pathways and clarify their impact on gene expression. RAW 264.7 cells were cultured for 5 days in DMEM supplemented with 10% FCS, 50 U/ml penicillin, 50 ug/ml streptomycin, and 1mM sodium pyruvate at 37C in a 5% CO2 in air incubator. Cells were washed, transferred into new flasks, and rested for 16 hr before being stimulated with 3.2 ug/ml CpG ODN and/or 32 ug/ml poly(I:C) for 4 - 12 hr. All experiments were independently repeated three times, and data from all experiments combined for analysis. Control samples included RAW 264.7 cells that were unstimulated and collected at 4 and 12 hours. All arrays used reverse transcribed mRNA from stimulted and unstimulated RAW cells as well as Universal Mouse Reference RNA purchased from Stratagene, La Holla, CA.

2009-07-21 | E-GEOD-15416 | biostudies-arrayexpress

Type I Interferon Dependent Anti-viral Activity

Project description:Synthetic oligonucleotides (ODN) expressing CpG motifs trigger an innate immune response via TLR9. Multiple microarray analyses were performed to identify the global effects on gene expression of stimulating the CAL-1 human pDC line with different types of CpG ODN. Results show that a subset of genes characterized by shared anti-viral activity was consistently up-regulated by ODNs that otherwise mediate discrete functions. This group of genes was largely dependant on autocrine type I interferon (IFN) signaling, as their induction was blocked by neutralizing antibody targeting the type I IFN receptor. Coupling these experiments with a meta-analysis of other published works led to the identification of a set of 32 functionally conserved genes that was reproducibly activated by different types of CpG DNA in different species and cell types. Functionally, these 'core' genes support a type I IFN response to viral infection, and differ from genes up-regulated by only a single type of CpG ODN. These findings help define the conserved and sequence-specific patterns of gene activation triggered via TLR9 and improve our understanding of the immunomodulatory effects elicited by CpG ODN.

2012-07-01 | GSE30849 | GEO

IGROV1 miRNA expression analysis after in vivo locoregional treatment with CpG-ODN

2013-03-31 | GSE41783 | GEO

Profiling of aPL response in mouse monocytes

Project description:We profiled CD115-selected mouse spleen monocytes after stimulation with aPL HL7G and after stimulation with Tlr7 agonist R848, Tlr9 agonist CpGB and Tlr3 agonist Poly I/C

2020-11-17 | GSE161575 | GEO

Project description:Toll-like receptors (TLRs) are mediators in the innate immune response by recognizing nucleic acids derived from microbial components as for TLR9 binding ability for CpG rich bacterial DNA. We studied the differential expression by microarrays of all known T-UCRs and near genes in a series of 6 primary B-CLL cases cells comparing cells treated in vitro with CpG ODN versus mock controls.

2018-12-31 | GSE57160 | GEO

Gene expression profiling of IGROV1 cells after in vitro treatment with ascitic fluid from human ovarian cancer bearing mice

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IGROV1 miRNA expression analysis after in vivo locoregional treatment with CpG-ODN

Project description:We reported that peri-tumoral CpG-ODN treatment, probably activating TLR9-expressing cells present in the tumor microenvironment, sensitized cancer cells to DNA-damaging chemotherapy (Cancer Res 2011 Oct 15;71(20):6382-90). Here, we investigated whether this treatment induces a modulation of miRNAs and their involvement in chemotherapy sensitivity. Twenty miRNAs were found differentially expressed in tumors from CpG-ODN-treated mice versus controls. Evaluation of the role of miR-424, miR-340 and miR-302b on cisplatin sensitivity revealed that ectopic expression of miR-302b (up-modulated in our array) in IGROV1 cells significantly improved cisplatin activity. The identification of miRNAs able to modify sensitivity to chemotherapy treatment will provide an experimental base for its future possible use as a target or tool of specific therapies. IGROV1 human ovarian carcinoma cells were adapted to growth intraperitoneally (i.p.) and maintained by serial i.p. passages of ascitic cells into healthy mice. Mice were injected i.p. with 2.5 x 10^6 ascitic cells in 0.2 ml of saline and treated starting 10-11 days later, when mice showed evident and established ascites, with CpG-ODN [phosphorothioated ODN1826 (5'-TCCATGACGTTCCTGACGTT-3')] delivered i.p. at a dose of 20 µg/mouse for 3 consecutive days, control mice received saline (4 mice/group). 24 hours after the last treatment, ascites-bearing mice were sacrificed. Tumor cells adherent to peritoneal wall were collected and immediately frozen in liquid nitrogen until RNA extraction.

2013-03-31 | E-GEOD-41783 | biostudies-arrayexpress

Gene expression profile of IGROV1 cells after in vitro treatment with CpG-ODN

Project description:CpG-ODN is a potent immuno-stimulatory molecule. In order to exclude a direct effect of murine CpG-ODN on IGROV1 human ovarian cancer cell line a gene expression experiment was performed.

2011-04-01 | GSE23442 | GEO

IGROV1 gene expression analysis after in vivo locoregional treatment with CpG-ODN

Project description:Toll-like receptor 9 synthetic agonist oligodeoxynucleotides expressing CpG motifs are currently evaluated for their anti-tumor activity, mainly in association with DNA-damaging drugs. Microarray expression analyses of genes implicates in DNA repair on tumor cells from mice treated with CpG-OD, revealed a down-regulation in tumor cells. These findings provide the first time evidence that immune cells upon TLR9 engagement can sensitize cancer cells to DNA damaging chemotherapy. IGROV-1 human ovarian carcinoma cells were adapted to growth intraperitoneally (i.p.) and maintained by serial i.p. passages of ascitic cells into healthy mice as described. Mice were injected i.p. with 2.5 x 106 ascitic cells in 0.2 ml of saline and treated starting 10-11 days later, when mice showed evident and established ascites, with CpG-ODN [phosphorothioated ODN1826 (5’-TCCATGACGTTCCTGACGTT-3’), TriLink Biotechnologies (San Diego, CA, USA)]. delivered i.p. at a dose of 20 µg/mouse for 3 consecutive days or 1 time, control mice received saline (4 mice/group). 24 hours after the last treatment with saline or CpG-ODN, ascites-bearing mice were sacrificed by cervical dislocation. Tumor cells adhered to peritoneal wall were collected and extraction immediately frozen in liquid nitrogen until RNA extraction.

2011-04-01 | E-GEOD-23441 | biostudies-arrayexpress

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Project description:The innate immune system is the first line of defense against microbial pathogens. The activated innate immune system plays important roles in eliciting antimicrobial defenses. Despite the benefits of innate immune responses, excessive inflammation will cause host damage. Thus, tight regulation of these processes is required for the maintenance of immune homeostasis. Recently, a new class of long non-coding RNAs (lncRNAs) has emerged as important regulators in many physiological and pathological processes. Dysregulated lncRNAs have been found to be associated with excessive or uncontrolled inflammation. In this study, we employed a lncRNA microarray-based profiling assay to detect changes of lncRNAs at different stages of CpG ODN-induced macrophage activation. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were performed based on the function of mRNAs. Co-expression and network potential targeting relationship were constructed according to the microarray results and bioinformatics predictions. Our findings revealed the involvement of lncRNAs in the process of CpG ODN-induced macrophage activation.

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