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Transcriptomic profile of murine macrophage RAW264.7 in response to Salmonella flagellin

ABSTRACT: Macrophages are an important component of the innate immune system. Flagellin has been extensively studied for its adjuvant activity owing to its TLR5 and NLRC4 binding sites. However, few studies have comprehensively investigated the effects of flagellin in macrophages using transcriptome sequencing. In this study, RNA sequencing (RNA-seq) was used to analyze the expression patterns of RAW264.7 cells induced by flagellin (FliC) of Salmonella typhimurium compared with unstimulated cells, to identify novel transcriptomic signatures in macrophages.

ORGANISM(S): Mus musculus

PROVIDER: GSE200796 | GEO | 2022/12/31

REPOSITORIES: GEO

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Cytosolic flagellin receptor NLRC4 protects mice against mucosal and systemic challenges

Project description:Bacterial flagellin is a dominant innate immune activator of the intestine. Therefore, we examined the role of the intracellular flagellin receptor, NLRC4, in protecting the gut and/or driving inflammation. In accord with NLRC4 acting via transcription-independent pathways, loss of NLRC4 did not reduce the rapid robust changes in intestinal gene expression induced by flagellin administration. Loss of NLRC4 did not alter basal intestinal homeostasis nor predispose mice to development of colitis upon administration of an anti-IL-10R monoclonal antibody. However, in response to epithelial injury induced by dextran sulfate sodium (DSS), loss of NLRC4 resulted in more severe disease indicating a role for NLRC4 in protecting the gut. Moreover, loss of NLRC4 resulted in increased mortality in response to flagellate, but not aflagellate Salmonella infection. Thus, despite not being involved in rapid intestinal gene remodeling upon detection of flagellin, NLRC4-mediated inflammasome activation protects mice from mucosal and systemic challenges Flagellin (FliC) from WT Salmonella enterica serovar Typhimurium (SL3201, fljB–) was purified through sequential cation and anion-exchange chromatography and purity was verified as previously described 4. WT, T5KO, N4KO and T5/N4-DKO mice (n=6) were given either 0.2 mL PBS or flagellin (10μg/mouse in 0.2 mL PBS) intraperitoneally. After 1h, mice were euthanized and colon was taken and stored in RNAlater (Invitrogen) for 1 day. Total mRNA was isolated from colonic tissues using TRIzol (Invitrogen) and purified using the RNeasy® Plus Mini kit (Qiagen) according to the manufacturer’s instructions

2012-01-01 | E-GEOD-34492 | biostudies-arrayexpress

Analysis of microRNA expression in murine BMDCs activated by flagellin

Project description:MicroRNAs (miRNAs) are small non-coding RNA molecules that regulate gene expression by either translational inhibition or mRNA degradation. However, the role of miRNAs in inflammation induced by flagellin (ligand of TLR5) has not been fully determined, Here, we identified differentially expressed miRNAs in murine bone marrow-derived dendritic cells (BMDCs) between flagellin (FliC) treatment and medium alone using miRNA microarray. And we further evaluated the role of miRNAs in inflammation induced by flagellin.

2021-11-13 | GSE188656 | GEO

Cytosolic flagellin receptor NLRC4 protects mice against mucosal and systemic challenges

2012-01-01 | GSE34492 | GEO

Gene expression induced by flagellin stimulation in IECs

Project description:Microarray analysis of IECs from Tlr5+/+ and Tlr5–/– mice stimulated with either medium alone or flagellin (1 µg/ml); to elucidate TLR5-mediated immune responses in IECs. Keywords: ordered

2006-08-01 | GSE5112 | GEO

Prevention and cure of rotavirus infection via TLR5/NLRC4-mediated production of IL-22 and IL-18

Project description:Activators of innate immunity may have potential to combat a broad range of infectious agents. We report that treatment with bacterial flagellin prevented rotavirus (RV) infection in mice and cured chronically RV-infected mice. Protection was independent of adaptive immunity and interferon (IFN, type I and II) and required flagellin receptors Toll-like receptor 5 (TLR5) and NOD-like receptor C4 (NLRC4). Flagellin-induced activation of TLR5 on dendritic cells elicited production of the cytokine interleukin (IL)-22, which induced a protective gene expression program in intestinal epithelial cells. Flagellin also induced NLRC4-dependent production of IL-18 and immediate elimination of RV-infected cells. Administration of IL-22 and IL-18 to mice fully recapitulated the capacity of flagellin to prevent or eliminate RV infection, and thus holds promise as a broad-spectrum antiviral agent.

2014-11-30 | GSE62479 | GEO

Gene expression induced by flagellin stimulation in CD11c+ LPCs

Project description:Microarray analysis of IECs from Tlr5+/+ and Tlr5-/- mice stimulated with either medium alone or flagellin (1 µg/ml); to elucidate TLR5-mediated immune responses in CD11c+ LPCs Keywords: ordered

2006-08-01 | GSE5113 | GEO

Transcriptome of FliC-stimulated mouse bladder and dependency on TLR5

Project description:We report a transcriptional response in the mouse bladder following exposure to the major flagella filament protein, FliC. The response is shown to encompasses multiple innate immune networks not previously associated with FliC. Additionally, we provide analysis of components of this response that depend on the presence of the known flagellin receptor, TLR5.

2019-06-25 | GSE132294 | GEO

IRF8 regulates transcription of Naips for NLRC4 inflammasome activation

Project description:Inflammasome activation is critical for host defense against various microbial infections. Activation of the NLRC4 inflammasome requires detection of flagellin or type III secretion system (T3SS) components by NLR family apoptosis inhibitory proteins (NAIPs); yet how this pathway is regulated is unknown. Here we found that interferon regulatory factor 8 (IRF8) is required for optimal activation of the NLRC4 inflammasome in bone marrow-derived macrophages infected with Salmonella Typhimurium, Burkholderia thailandensis, or Pseudomonas aeruginosa but is dispensable for activation of the canonical and non-canonical NLRP3, AIM2, and Pyrin inflammasomes. IRF8 governs the transcription of Naips to allow detection of flagellin or T3SS proteins to mediate NLRC4 inflammasome activation. Furthermore, we found that IRF8 confers protection against bacterial infection in vivo, owing to its role in inflammasome-dependent cytokine production and pyroptosis. Altogether, our findings suggest that IRF8 is a critical regulator of NAIPs and NLRC4 inflammasome activation for defense against bacterial infection.

2018-04-27 | GSE110452 | GEO

Transcription profiling of muse flagellin stimulated CD11c+ LPCs

Project description:Microarray analysis of IECs from Tlr5+/+ and Tlr5â/â mice stimulated with either medium alone or flagellin (1 Âµg/ml); to elucidate TLR5-mediated immune responses in CD11c+ LPCs Experiment Overall Design: CD11c+ LPCs from Tlr5+/+ and Tlr5â/â mice were treated with or without flagellin (1 ïg/ml) for 4 h. Total RNA was extracted with an RNeasy kit (Qiagen), and purified using an Oligotex mRNA Kit (Pharmacia). Fragmented and biotin-labeled cDNA was synthesized from 100 ng purified mRNA using the Ovation Biotin System (Nugen), according to the manufacturerâs protocol. cDNA was hybridized to Affymetrix Murine Genome 430 2.0 microarray chips (Affymetrix) according to the manufacturerâs instructions. Hybridized chips were stained, washed and scanned using a GeneArray Scanner (Affymetrix). Data analysis was performed with Microarray Suite software (Version 5.0, Affymetrix) and GeneSpring software (Silicon Genetics).

2008-06-13 | E-GEOD-5113 | biostudies-arrayexpress

Transcription profiling of mouse IECS response to flagellin stimulation

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2008-06-13 | E-GEOD-5112 | biostudies-arrayexpress

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