IL-17 primes airway epithelial cells lacking functional Cystic Fibrosis Transmembrane conductance Regulator (CFTR) to increase NOD1 responses

doi:10.1016/j.bbrc.2009.11.088

Biochemical and Biophysical Research Communications

Volume 391, Issue 1, 1 January 2010, Pages 505-509

https://doi.org/10.1016/j.bbrc.2009.11.088 Get rights and content

Abstract

In Cystic Fibrosis (CF), the absence of functional Cystic Fibrosis Transmembrane conductance Regulator (CFTR) translates into chronic bacterial infection, excessive inflammation, tissue damage, impaired lung function and eventual death. Understanding the mechanisms underlying this vicious circle of inflammation is key to better therapies for CF. In this manuscript, we have found that the presence of IL-17 in the airways of CF patients not only exacerbates inflammation through the recruitment of neutrophils via secretion of CXCL8, but also by priming airway epithelial cells lacking functional CFTR to increase response to the bacterial sensor NOD1. IL-17 stimulation of airway epithelial cells (AECs) lacking functional CFTR increased the expression of NOD1, NOD2, TLR4 and its own receptors IL-17RA and IL-17RC. Moreover, prior stimulation of AECs expressing the CFTRΔF508 mutant with IL-17 showed much greater CXCL8 secretion in response to a NOD1 agonist and Pseudomonas aeruginosa diffusible material. Taken together our results show that IL-17 primes AECs expressing CFTRΔF508 to increase host defence response to bacteria through the up-regulation of PRRs, and in particular of NOD1, and identifies another mechanism of action through which the CFTRΔF508 mutation leads to increase inflammation in response to bacterial ligands. Therefore preventing IL-17 function in CF may prove an important strategy in decreasing lung inflammation due to both direct and indirect effects.

Section snippets

Background

Cystic Fibrosis is one of the most common fatal genetic diseases affecting Caucasians of European decent. It is an autosomal recessive disorder caused by mutations of the CF transmembrane conductance regulator (CFTR). Defective CFTR function in the airway epithelium is responsible for CF lung disease, the most life-threatening complication of CF, characterized by mucus hypersecretion and neutrophil-dominated inflammation. Pseudomonas aeruginosa (PA) infections occur in 70% of individuals at an

Materials and methods

Materials. PD184352 was bought from USBiological (Swampscott, MA, USA). BIRB07896 was kindly provided by Professor Sir Philip Cohen (MRC PPU, University of Dundee, UK). Recombinant human IL-17A was purchased from Biovision (#4176-25; CA, USA). LPS from P. aeruginosa and C12-ie-DAP were bought from Invivogen (CA, USA). The diffusible PA material was prepared as previously reported [13]. Briefly, filtrates of late stationary phase PA (mucoid strain 508) were used. PA 508 is a stable mucoid

IL-17 up-regulates NOD1, NOD2, IL-17RA, IL-17RC and TLR4 expression in AECs lacking functional CFTR expression

In order to have a better understanding of the role of IL-17 in cystic fibrosis, we stimulated AECs expressing wild-type (NuLi) and ΔF508 (CuFi) CFTR with IL-17A and measured changes in gene expression. As previously reported [8], IL-17 up-regulated CXCL8 mRNA as determined by real-time PCR in a p38 MAPK and ERK1/ERK2-dependent manner (Fig. 1A). Although the effects of IL-17 on chemokine and cytokine gene expression have been studied, much less is known about its potential effects on the

Conclusion

In this manuscript we have discovered an unexpected function of IL-17 in the CF lungs, which is to prime AECs expressing CFTRΔF508 to increase host defence response to bacteria, like the well known CF pathogen P. aeruginosa, through the up-regulation of PRR and in particular of NOD1. Therefore preventing IL-17 function in CF may prove an important strategy in decreasing lung inflammation due to both direct and indirect effects. An important question that remains is how the presence of the

Acknowledgments

We would like to thank Sir Philip Cohen (MRC PPU, University of Dundee, UK) for the kind gift of BIRB0796. The authors declare no conflict of interest. We acknowledge the financial support of the Department of Medicine, McGill University and the McGill University Health Centre Research Institute (MUHC-RI) and the Canadian Foundation for Innovation-leaders opportunities funds. The Meakins-Christie Laboratories – MUHC-RI, are supported by a Centre grant from Les Fonds de la Recherche en Santé du

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Multi-omic comparisons between CFBE41o- cells stably expressing wild-type CFTR and F508del-mutant CFTR
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Citation Excerpt :
Researchers have shown that IL-17A and IL-17F are elevated in the P. aeruginosa colonized sputum of CF patients during pulmonary exacerbation and are reduced under antibiotic therapy [23]. In addition, early inflammation in the CF lung is potentially initiated by Th17 cells, which secrete IL-17 to promote neutrophil recruitment and prime the CFTR dysfunctional airway epithelial cells into a hyperinflammatory state [24,25]. Our mRNA-seq analyses showed that both WTBE and CFBE cells upregulated the IL-17 signaling pathway (Rank 1 of 10) and Th17 cell differentiation pathway (Rank 5 of 10) during polarization, but upregulation scale was greater in CFBE cells.
Cystic fibrosis (CF) is characterized by chronic inflammation and excessive cytokines secretion in the lung. Isogenic human CF bronchial epithelial (CFBE41o-) cell lines stably expressing wt-CFTR (WTBE) or F508del mutant (CFBE) are widely used tools in understanding responses to stimuli or drugs and CF pathogenesis in vitro. However, the intrinsic cellular differences in culture are unknown.
We performed integrative analyses of these isogenic cells at the protein, mRNA, and chromatin levels in the submerged and air-liquid interface (ALI) conditions to determine cell intrinsic effects of mutant versus complemented CFTR expression.
CFBE and WTBE cells displayed different cytokine secretion patterns, including IL-6, IL-8, CXCL1, CXCL10, and CCL5. The ALI culture dramatically increased cytokine secretion in both cells. Assay for transposase-accessible chromatin using sequencing (ATAC-seq) result showed different chromatin landscapes upon polarization and CFBE cells, compared to WTBE cells, exhibited higher genome-wide chromatin accessibility under both culture methods. At the transcriptome level, differentially expressed genes identified by mRNA sequencing between two cell lines were highly concentrated in immunity-related pathways.
This multilayered study shows that expression of wild-type CFTR has an epithelial cell intrinsic effect on the cell's epigenome and transcriptome particularly in immunity relevant activities. These data will serve as a resource for the CF community and may serve as epithelial biomarkers for CFTR mRNA therapy.
Could T cells be involved in lung deterioration and hyperglycemia in cystic fibrosis?
2014, Diabetes Research and Clinical Practice
Citation Excerpt :
CFRD is an important complication of CF that is preceded by a long pre-diabetic phase associated with unexplained accelerated loss of lung functions [16]. There is some evidence that Th17 cells and dysfunctional Treg lymphocytes producing IL-17 could be implicated both in lung damage, perpetuating the excessive pulmonary inflammatory process, and in β-cell dysfunction, providing a potential link between these two key and frequent abnormalities in CF [38–41,44–51]. Lymphocytes are susceptible to hyperglycemia [68,69] but also to non-optimal levels of vitD which could both trigger the abnormal immune response observed in CF [68,69,78–80].
Cystic fibrosis-related diabetes (CFRD) is the most frequent complication of cystic fibrosis (CF) and associated with increased mortality. Why patients have an accelerated loss of lung function before the diagnosis of CFRD remains poorly understood. We reported that patients with or without CFRD had increased glucose excursions when compared to healthy peers. Studies have demonstrated that patients with CF have increased glucose fluctuations and hyperglycemia and that this may affect the clinical course of CF and lead to lymphocyte dysfunction. T-helper 17 (Th17) lymphocytes produce and secrete the pro-inflammatory cytokine IL-17. The Th17 pathway is involved in CF lung inflammation, β-cell destruction in type 1 diabetes (T1D) and Th17 cells of patients with type 2 diabetes have increased production of IL-17 when compared to healthy peers. Also, regulatory T-cells (Tregs) have been shown to be dysfunctional and produce IL-17 in T1D. Furthermore, vitamin D can affect inflammation in CF, diabetes and the differentiation of lymphocytes. In this review, we discuss the potential roles of hyperglycemia on Th17 cells, Tregs and IL-17 as a potential cause for accelerated lung function decline before CFRD and how this could be modulated by vitamin D or by directly intervening in the IL-17A pathway.
Synergism between interleukin (IL)-17 and toll-like receptor 2 and 4 signals to induce IL-8 expression in cystic fibrosis airway epithelial cells
2012, Journal of Pharmacological Sciences
Cystic fibrosis (CF) is the most common lethal inherited disorder and is caused by mutations in the gene encoding the CF transmembrane regulator (CFTR). The CF lung expresses a profound proinflammatory phenotype that appears to be related to a constitutive hypersecretion of interleukin (IL)-8 from airway epithelial cells in response to microbial infection. Since overproduction of IL-8 in CF contributes to massive bronchial infiltrates of neutrophils, identification of the pathways underlying IL-8 induction could provide novel drug targets for treatment of neutrophil-dominated inflammatory diseases such as CF. Here, we show that IL-17A synergistically increases IL-8 production induced by a toll-like receptor (TLR) 2 agonist, peptidoglycan (PGN), or TLR4 agonist, lipopolysaccharide (LPS), in a human CF bronchial epithelial cell line (CFBE41o-). A strong synergism was also observed in primary human CF bronchial epithelial cells, but not in human non-CF cell lines and primary cells. Notably, despite the induction of nuclear factor-κB and MAP kinases during TLR2 or TLR4 activation in CFBE41o-, IL-17A-dependent synergism appears to be the result of enhanced PGN- or LPS-induced phosphorylation of p38. Taken together, these studies provide evidence that IL-17A is a critical factor in increasing IL-8 expression in bacteria-infected CF airways via a pathway that regulates p38 phosphorylation.
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2022, Thorax
Exosomes derived from human adipose mesenchymal stem cells attenuate hypertrophic scar fibrosis by miR-192-5p/IL-17RA/Smad axis
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Nucleotide-oligomerizing domain-1 activation exaggerates cigarette smoke-induced chronic obstructive pulmonary-like disease in mice
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IL-17 primes airway epithelial cells lacking functional Cystic Fibrosis Transmembrane conductance Regulator (CFTR) to increase NOD1 responses

Abstract

Section snippets

Background

Materials and methods

IL-17 up-regulates NOD1, NOD2, IL-17RA, IL-17RC and TLR4 expression in AECs lacking functional CFTR expression

Conclusion

Acknowledgments

Chest

J. Biol. Chem.

Curr. Opin. Immunol.

Immunity

J. Allergy Clin. Immunol.

Cell. Signal.

Lung infections associated with cystic fibrosis

Clin. Microbiol. Rev.

Nod-like proteins in immunity, inflammation and disease

Nat. Immunol.

Role of IL-17A, IL-17F, and the IL-17 receptor in regulating growth-related oncogene-alpha and granulocyte colony-stimulating factor in bronchial epithelium: implications for airway inflammation in cystic fibrosis

J. Immunol.