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Genome-wide transcriptional analysis of flagellin induced reprogramming in mouse corneal epithelial cells in response to Pseudomonas aeruginosa

ABSTRACT: We previously showed that pre-exposure of the cornea to TLR5 ligand flagellin induces profound mucosal innate protection against pathogenic microbes by reprogramming gene expression. To date, there was no genome-wide cDNA array to detect full scale of flagellin mediated reprogramming of gene expression in mucosal surface epithelial cells. Taking advantage of readily accessible, easily procurable epithelial cell population, this study is the first report to use genome-wide cDNA microarray approach to document genes associated with flagellin-induced protection against Pseudomonas aeruginosa infection in corneal epithelial cells (CECs).

ORGANISM(S): Mus musculus

PROVIDER: GSE41147 | GEO | 2013/01/01

SECONDARY ACCESSION(S): PRJNA175920

REPOSITORIES: GEO

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Genome-wide transcriptional analysis of flagellin induced reprogramming in mouse corneal epithelial cells in response to Pseudomonas aeruginosa

Project description:We previously showed that pre-exposure of the cornea to TLR5 ligand flagellin induces profound mucosal innate protection against pathogenic microbes by reprogramming gene expression. To date, there was no genome-wide cDNA array to detect full scale of flagellin mediated reprogramming of gene expression in mucosal surface epithelial cells. Taking advantage of readily accessible, easily procurable epithelial cell population, this study is the first report to use genome-wide cDNA microarray approach to document genes associated with flagellin-induced protection against Pseudomonas aeruginosa infection in corneal epithelial cells (CECs). Total RNA obtained from isolated mouse corneal epithelial cells of the control (cells scrapped off from the corneas without infection), Pseudomonas aeruginosa infected (6 h post infection) and flagellin pretreated (24 h), followed by Pseudomonas aeruginosa infection (6 h).

2013-01-01 | E-GEOD-41147 | biostudies-arrayexpress

Mouse lung transcriptional response to flagellin stimulation

Project description:Vaccine adjuvants enhance adaptive immunity to co-administered antigens. Whereas the modes of action are multiple, the activation of antigen-presenting cells (APC) like dendritic cells by adjuvants is a prerequisite. Detection of microbial signals by innate sensors like Toll-like receptors (TLR) is a major mechanism of APC activation. Most candidate or licensed vaccines assume that adjuvant activity of TLR agonists depends on direct effect on APCs. This study addressed whether TLR stimulation of non-hematopoietic cells could contribute to the adjuvant effect. Nasal administration of flagellin enhanced T cell- and antibody-mediated immunity to co-administered antigens in a TLR5-dependent but inflammasome-independent manner. We found that lung radioresistant cells were sufficient to promote immunity, thereby suggesting that direct TLR5-mediated APC stimulation is dispensable to adjuvant activity. Consistent with this, radioresistant compartment is essential to stimulate the swift TLR5-dependent transcription. The transcriptional response was restricted to the epithelial compartment and was associated to the production of a narrow set of mediators including the chemokine CCL20, known to promote APC recruitment in mucosal tissues. Besides, flagellin was rapidly degraded in lower airways and was not transported into lung parenchyma or peripheral tissues. This study therefore suggests an unexpected mechanism for how TLR agonists act as adjuvant and how epithelium is instrumental to sense and integrate microbial signals to promote adaptive immunity. In conclusion, the immune-enhancing effect of adjuvants on epithelial cells can be harnessed for improving vaccines. 1 µg of flagellin was instillated intranasally to LPS-unresponsive C3H/HeJ mice. Total lung RNA was extracted 1h later and prepare for hybridization on Affymetrix microarrays.

2013-03-22 | E-GEOD-45369 | biostudies-arrayexpress

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

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 IECs. Experiment Overall Design: IECs 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-5112 | biostudies-arrayexpress

Lung conventional dendritic cells promote mucosal adjuvant activity of flagellin

Project description:The activity of the potent nasal adjuvant flagellin depends on initial activation of airway epithelilal cells. This study aims at investigating the immune cells involved in the antigen presentation within the respiratory tract. First, microarrays characterized that activation/recruitment of immune cells was the main signature of flagellin activation in lung. Neutrophils and classical monocytes were strongly recruited into the lungs and take up antigen upon nasal administration of flagellin. In contrast, the numbers of lung CD11b+ or CD103+ DC and alveolar macrophages were not enhanced although these cells were very efficient in antigen uptake. Our experiments showed that CD11b+ DC but not monocytes, PMN, CD103+ DC are not essential for the adjuvant effect. Flagellin signaling enhanced the functional activation of lung CD11b+ conventional DC and their migration to the lymph nodes. Using Cd11c-DTR mice, CD11b+ DC from lymph nodes were identified as the major stimulators of CD4 T cell activation. Finally, the migration and maturation of mucosal DC was independent of direct signaling, highlighting the contribution of epithelial factors in the mucosal adjuvant effect of flagellin. In conclusion, these data demonstrate that adjuvant activity can be dissociated from inflammatory cell recruitment an open new perpectives for improvement of vaccines. Female C57BL/6J (6 weeks old) mice were immunized with the TLR5 agonist Flagellin (1Î¼g diluted in PBS) by intranasal (i.n.) administration. Blood samples of four time points post immunization were taken at 2h, 4h, 18h and 48h. Microarray experiments were performed as single-color hybridizations.

2014-01-01 | E-GEOD-46421 | biostudies-arrayexpress

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

Mouse lung transcriptional response to flagellin stimulation

2013-03-22 | GSE45369 | 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

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

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

Project description: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.

2022-12-31 | GSE200796 | GEO

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