scholarly journals Cystic Fibrosis Defective Response to Infection Involves Autophagy and Lipid Metabolism

Cells ◽  
2020 ◽  
Vol 9 (8) ◽  
pp. 1845
Author(s):  
Alessandra Mingione ◽  
Emerenziana Ottaviano ◽  
Matteo Barcella ◽  
Ivan Merelli ◽  
Lorenzo Rosso ◽  
...  
Keyword(s):  
Cystic Fibrosis ◽  
Lipid Metabolism ◽  
Cell Line ◽  
Bronchial Epithelial Cell ◽  
Hereditary Disease ◽  
Multiple Organ ◽  
Epithelial Cell Line ◽  
Blood Monocytes ◽  
Microbial Infection ◽  
Pathogen Clearance

Cystic fibrosis (CF) is a hereditary disease, with 70% of patients developing a proteinopathy related to the deletion of phenylalanine 508. CF is associated with multiple organ dysfunction, chronic inflammation, and recurrent lung infections. CF is characterized by defective autophagy, lipid metabolism, and immune response. Intracellular lipid accumulation favors microbial infection, and autophagy deficiency impairs internalized pathogen clearance. Myriocin, an inhibitor of sphingolipid synthesis, significantly reduces inflammation, promotes microbial clearance in the lungs, and induces autophagy and lipid oxidation. RNA-seq was performed in Aspergillusfumigatus-infected and myriocin-treated CF patients’ derived monocytes and in a CF bronchial epithelial cell line. Fungal clearance was also evaluated in CF monocytes. Myriocin enhanced CF patients’ monocytes killing of A. fumigatus. CF patients’ monocytes and cell line responded to infection with a profound transcriptional change; myriocin regulates genes that are involved in inflammation, autophagy, lipid storage, and metabolism, including histones and heat shock proteins whose activity is related to the response to infection. We conclude that the regulation of sphingolipid synthesis induces a metabolism drift by promoting autophagy and lipid consumption. This process is driven by a transcriptional program that corrects part of the differences between CF and control samples, therefore ameliorating the infection response and pathogen clearance in the CF cell line and in CF peripheral blood monocytes.

scholarly journals Staphylococcus aureus Escapes More Efficiently from the Phagosome of a Cystic Fibrosis Bronchial Epithelial Cell Line than from Its Normal Counterpart

2006 ◽  
Vol 74 (5) ◽  
pp. 2568-2577 ◽  
Author(s):  
Todd M. Jarry ◽  
Ambrose L. Cheung
Keyword(s):  
Cystic Fibrosis ◽  
Staphylococcus Aureus ◽  
Epithelial Cell ◽  
Cell Line ◽  
Cell Lines ◽  
Microscopic Analysis ◽  
Bronchial Epithelial Cell ◽  
Epithelial Cell Line ◽  
Similar Degree ◽  
Electron Microscopic

ABSTRACTStaphylococcus aureusis frequently the initial bacterium isolated from young cystic fibrosis (CF) patients, and yet its role in CF disease progression has not been determined. Recent data from our lab demonstrates thatS. aureuscan invade and replicate within the CF tracheal epithelial cell line (CFT-1). Here we describe the finding that the fate of internalizedS. aureusin CFT-1 cells differs from its complemented counterpart (LCFSN).S. aureusstrain RN6390 was able to replicate within the mutant CFT-1 cells after invasion but not in the complemented LCFSN cells. At 1 h postinvasion,S. aureuscontaining vesicles within both cell lines acquired vacuolar-ATPase, lysosomal markers LAMP 1 and 2, and the lysomotrophic dye LysoTracker to a similar degree. However, at 4 h postinvasion, the percentage ofS. aureuswithin CFT-1 cells associated with these markers decreased significantly compared to LCFSN, where the association approached 100%. Transmission electron microscopic analysis revealed that the majority of bacteria within CFT-1 cells were free in the cytosol at 4 h after invasion, whereas mostS. aureusbacteria internalized by LCFSN cells remained within vesicles. These results demonstrate a fundamental difference in the fate of liveS. aureusafter invasion of CFT-1 versus LCFSN cell lines and may explain the propensity ofS. aureusto cause chronic lung infection in CF patients.

Glycosylation differences between a cystic fibrosis and rescued airway cell line are not CFTR dependent

1997 ◽  
Vol 273 (5) ◽  
pp. L913-L920 ◽  
Author(s):  
Xiaosui Jiang ◽  
Warren G. Hill ◽  
Joseph M. Pilewski ◽  
Ora A. Weisz
Keyword(s):  
Cystic Fibrosis ◽  
Cell Surface ◽  
Cell Line ◽  
Cell Lines ◽  
Recombinant Adenovirus ◽  
Lectin Binding ◽  
Bronchial Epithelial Cell ◽  
Epithelial Cell Line ◽  
Binding Assays ◽  
The Difference

Altered glycosylation of mucus and membrane glycoconjugates could explain reported differences in binding of bacterial pathogens to cystic fibrosis (CF) versus normal tissue. However, because bacteria can alter cell surface glycoconjugates, it is not possible to assess the role of cystic fibrosis transmembrane conductance regulators (CFTR) in glycosylation in these studies. To address this issue, we have developed quantitative lectin binding assays to compare cell surface glycosylation in well-matched immortalized CF cells and rescued cell lines. The CF airway bronchial epithelial cell line IB3-1 consistently bound more peanut agglutinin (PNA) than its clonal derivative S9, which stably expresses functional wild-type CFTR. Pretreatment with neuraminidase increased PNA binding and abolished the difference between the two cell lines. However, infection of the IB3-1 cells with a replication-deficient recombinant adenovirus encoding CFTR restored CFTR function but did not alter PNA binding to cells. In contrast, treatment with the weak base ammonium chloride increased PNA binding to both cell lines as expected. Our data show that even clonally related CF and rescued cells can exhibit significant differences in carbohydrate processing. Although the differences that we found are consistent with the proposed role for CFTR in modulating intraorganellar pH, our data strongly suggest that they are CFTR independent. These studies add a cautionary note to the interpretation of differences in glycosylation between CF and normal primary tissues and immortalized cells.

scholarly journals A Disease-Relevant High-Content Screening Assay to Identify Anti-Inflammatory Compounds for Use in Cystic Fibrosis

2010 ◽  
Vol 15 (10) ◽  
pp. 1204-1210 ◽  
Author(s):  
Angela M. Giddings ◽  
Rangan Maitra
Keyword(s):  
Cystic Fibrosis ◽  
Cell Line ◽  
Lung Inflammation ◽  
High Throughput Screening ◽  
Fluorescent Protein ◽  
Bronchial Epithelial Cell ◽  
Epithelial Cell Line ◽  
Screening Assay ◽  
Gfp Reporter ◽  
Anti Inflammatory

Chronic lung inflammation caused by bacterial pathogenesis through activation of nuclear factor kappa B (NFκB)–responsive proinflammatory genes is a major hurdle in the management of lung disease in cystic fibrosis (CF) patients. The authors generated a disease-relevant cell-based high-content screen to identify novel anti-inflammatory compounds for treating lung inflammation in CF. The human bronchial epithelial cell line KKLEB, harboring the most common form of mutation that causes CF, was modified to express an NFκB-responsive green fluorescent protein (GFP) reporter. After creation, the cell line was tested for its ability to respond to disease-relevant inflammatory stimuli elicited by treatment of cells with filtrates of Pseudomonas aeruginosa isolated from the airways of a CF patient. P. aeruginosa filtrates potently activated NFκB-responsive GFP reporter expression in cells. Subsequently, the assay was optimized for high-throughput screening (HTS) through generation of a Z factor (~0.5) and by testing its tolerance to the commonly used solvents ethanol and DMSO. A pilot library of clinically approved compounds was screened for assay validation. Several compounds with known NFκB inhibitory activity were identified, including several steroidal compounds that have been clinically tested in CF. Thus, the assay can be used in a broader HTS campaign to find anti-inflammatory agents for use in CF.

Retinoic acid inhibits interleukin-4-induced eotaxin production in a human bronchial epithelial cell line

2004 ◽  
Vol 286 (4) ◽  
pp. L777-L785 ◽  
Author(s):  
Kei Takamura ◽  
Yasuyuki Nasuhara ◽  
Motoko Kobayashi ◽  
Tomoko Betsuyaku ◽  
Yoko Tanino ◽  
...  
Keyword(s):  
Retinoic Acid ◽  
Epithelial Cell ◽  
Cell Line ◽  
Nuclear Translocation ◽  
Bronchial Epithelial Cell ◽  
Inhibitory Effect ◽  
Human Bronchial Epithelial Cell ◽  
Epithelial Cell Line ◽  
Transcriptional Level ◽  
Bronchial Epithelial

Retinoic acid (RA) is known to accelerate wound healing and induce cell differentiation. All- trans RA (ATRA) exerts its effect by binding retinoic acid receptors, which are members of the nuclear receptor family. We investigated whether RA can alter expression of eotaxin, a potent eosinophil chemoattractant that is regulated by the transcription factors signal transducer and activator of transcription 6 (STAT6) and NF-κB. We examined the effects of RA on eotaxin expression in a human bronchial epithelial cell line BEAS-2B. ATRA and its stereodimer 9- cis retinoic acid (9- cis RA) inhibited IL-4-induced release of eotaxin at 10-6M by 78.0 and 52.0%, respectively ( P < 0.05). ATRA and 9- cis RA also significantly inhibited IL-4-induced eotaxin mRNA expression at 10-6M by 52.3 and 53.5%, respectively ( P < 0.05). In contrast, neither ATRA nor 9- cis RA had any effects on TNF-α-induced eotaxin production. In transfection studies using eotaxin promoter luciferase plasmids, the inhibitory effect of ATRA on IL-4-induced eotaxin production was confirmed at the transcriptional level. Interestingly, ATRA had no effects on IL-4-induced tyrosine phosphorylation, nuclear translocation, or DNA binding activity of STAT6. Activating protein-1 was not involved in ATRA-mediated transrepression of eotaxin with IL-4 stimulation. The mechanism of the inhibitory effect of ATRA on IL-4-induced eotaxin production in human bronchial epithelial cells has not been elucidated but does not appear to be due to an effect on STAT6 activation. These findings raise the possibility that RA may reduce eosinophilic airway inflammation, one of the prominent pathological features of allergic diseases such as bronchial asthma.

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