Adenovirus 5–Fiber 35 Chimeric Vector Mediates Efficient Apical Correction of the Cystic Fibrosis Transmembrane Conductance Regulator Defect in Cystic Fibrosis Primary Airway Epithelia

2010 ◽  
Vol 21 (3) ◽  
pp. 251-269 ◽  
Author(s):  
Ophélia Granio ◽  
Katherine J.D. Ashbourne Excoffon ◽  
Petra Henning ◽  
Patricia Melin ◽  
Caroline Norez ◽  
...  
Keyword(s):  
Cystic Fibrosis ◽  
Transmembrane Conductance Regulator ◽  
Transmembrane Conductance ◽  
Airway Epithelia ◽  
Cystic Fibrosis Transmembrane ◽  
Adenovirus 5

scholarly journals Infection by Toxoplasma gondii, a severe parasite in neonates and AIDS patients, causes impaired anion secretion in airway epithelia

2015 ◽  
Vol 112 (14) ◽  
pp. 4435-4440 ◽  
Author(s):  
Hong-Mei Guo ◽  
Jiang-Mei Gao ◽  
Yu-Li Luo ◽  
Yan-Zi Wen ◽  
Yi-Lin Zhang ◽  
...  
Keyword(s):  
Cystic Fibrosis ◽  
Toxoplasma Gondii ◽  
Infected Mouse ◽  
Inflammatory Responses ◽  
Short Circuit ◽  
Transmembrane Conductance Regulator ◽  
Transmembrane Conductance ◽  
Airway Epithelia ◽  
Anion Secretion ◽  
Cystic Fibrosis Transmembrane

The airway epithelia initiate and modulate the inflammatory responses to various pathogens. The cystic fibrosis transmembrane conductance regulator-mediated Cl− secretion system plays a key role in mucociliary clearance of inhaled pathogens. We have explored the effects of Toxoplasma gondii, an opportunistic intracellular protozoan parasite, on Cl− secretion of the mouse tracheal epithelia. In this study, ATP-induced Cl− secretion indicated the presence of a biphasic short-circuit current (Isc) response, which was mediated by a Ca2+-activated Cl− channel (CaCC) and the cystic fibrosis transmembrane conductance regulator. However, the ATP-evoked Cl− secretion in T. gondii-infected mouse tracheal epithelia and the elevation of [Ca2+]i in T. gondii-infected human airway epithelial cells were suppressed. Quantitative reverse transcription–PCR revealed that the mRNA expression level of the P2Y2 receptor (P2Y2-R) increased significantly in T. gondii-infected mouse tracheal cells. This revealed the influence that pathological changes in P2Y2-R had on the downstream signal, suggesting that P2Y2-R was involved in the mechanism underlying T. gondii infection in airways. These results link T. gondii infection as well as other pathogen infections to Cl− secretion, via P2Y2-R, which may provide new insights for the treatment of pneumonia caused by pathogens including T. gondii.

scholarly journals Transcytosis maintains CFTR apical polarity in the face of constitutive and mutation-induced basolateral missorting

2018 ◽  
Author(s):  
Aurélien Bidaud-Meynard ◽  
Florian Bossard ◽  
Andrea Schnúr ◽  
Ryosuke Fukuda ◽  
Guido Veit ◽  
...  
Keyword(s):  
Cystic Fibrosis ◽  
Pdz Domain ◽  
Transmembrane Conductance Regulator ◽  
Transmembrane Conductance ◽  
Airway Epithelia ◽  
Apical Polarity ◽  
The Face ◽  
Domain Protein ◽  
Polarized Delivery ◽  
Cystic Fibrosis Transmembrane

SUMMARYApical polarity of cystic fibrosis transmembrane conductance regulator (CFTR) is essential for solute and water transport in secretory epithelia and can be impaired in human diseases. Maintenance of apical polarity in the face of CFTR non-polarized delivery and compromised apical retention of mutant CFTRs lacking PDZ-domain protein (NHERF1) interaction, remains enigmatic. Here we show that basolateral CFTR delivery originates from biosynthetic (~35%) and endocytic (~65%) recycling missorting. Basolateral channels are retrieved via basolateral-to-apical transcytosis, enhancing CFTR apical expression by two-fold and suppressing its degradation. CFTR transcytosis is microtubule-dependent but independent of Myo5B-, Rab11- and NHERF1 binding to its C-terminal DTRL motif in airway epithelia. Increased basolateral delivery due to compromised apical recycling and accelerated internalization upon impaired NHERF1-CFTR association is largely counterbalanced by CFTR efficient basolateral internalization and apical transcytosis. Thus, transcytosis represents a previously unrecognized but indispensable mechanism for maintaining CFTR apical polarity by attenuating its constitutive and mutation-induced basolateral missorting.

scholarly journals Abnormal localization of cystic fibrosis transmembrane conductance regulator in primary cultures of cystic fibrosis airway epithelia.

1992 ◽  
Vol 118 (3) ◽  
pp. 551-559 ◽  
Author(s):  
G M Denning ◽  
L S Ostedgaard ◽  
M J Welsh
Keyword(s):  
Cystic Fibrosis ◽  
Plasma Membrane ◽  
Apical Membrane ◽  
Specific Binding ◽  
Primary Cultures ◽  
Transmembrane Conductance Regulator ◽  
Transmembrane Conductance ◽  
Airway Epithelia ◽  
Cystic Fibrosis Transmembrane ◽  
Cl Permeability

Cystic fibrosis (CF) is caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR), a membrane glycoprotein that forms Cl- channels. Previous work has shown that when some CF-associated mutants of CFTR are expressed in heterologous cells, their glycosylation is incomplete. That observation led to the hypothesis that such mutants are not delivered to the plasma membrane where they can mediate Cl- transport. Testing this hypothesis requires localization of CFTR in nonrecombinant cells and a specific determination of whether CFTR is in the apical membrane of normal and CF epithelia. To test the hypothesis, we used primary cultures of airway epithelia grown on permeable supports because they polarize and express the CF defect in apical Cl- permeability. Moreover, their dysfunction contributes to disease. We developed a semiquantitative assay, using nonpermeabilized epithelia, an antibody directed against an extracellular epitope of CFTR, and large (1 microns) fluorescent beads which bound to secondary antibodies. We observed specific binding to airway epithelia from non-CF subjects, indicating that CFTR is located in the apical membrane. In contrast, there was no specific binding to the apical membrane of CF airway epithelia. These data were supported by qualitative studies using confocal microscopy: the most prominent immunostaining was in the apical region of non-CF cells and in cytoplasmic regions of CF cells. The results indicate that CFTR is either missing from the apical membrane of these CF cells or it is present at a much reduced level. The data support the proposed defective delivery of some CF-associated mutants to the plasma membrane and explain the lack of apical Cl- permeability in most CF airway epithelia.

Effects of cystic fibrosis transmembrane conductance regulator and ΔF508CFTR on inflammatory response, ER stress, and Ca2+ of airway epithelia

2007 ◽  
Vol 293 (5) ◽  
pp. L1250-L1260 ◽  
Author(s):  
Kevin Hybiske ◽  
Zhu Fu ◽  
Christian Schwarzer ◽  
Jill Tseng ◽  
Jiun Do ◽  
...  
Keyword(s):  
Cystic Fibrosis ◽  
Er Stress ◽  
Primary Cultures ◽  
Transmembrane Conductance Regulator ◽  
Innate Immune Responses ◽  
Transmembrane Conductance ◽  
Airway Epithelia ◽  
Specific Effects ◽  
Er Retention ◽  
Cystic Fibrosis Transmembrane

We tested whether cystic fibrosis (CF) airway epithelia have larger innate immune responses than non-CF or cystic fibrosis transmembrane conductance regulator (CFTR)-corrected cells, perhaps resulting from ER stress due to retention of ΔF508CFTR in the endoplasmic reticulum (ER) and activation of cytosolic Ca2+ (Cai) and nuclear factor (NF)-κB signaling. Adenovirus infections of a human CF (ΔF508/ΔF508) nasal cell line (CF15) provided isogenic comparisons of wild-type (wt) CFTR and ΔF508CFTR. In the absence of bacteria, there were no or only small differences among CF15, CF15-lacZ (β-galactosidase-expressing), CF15-wtCFTR (wtCFTR-corrected), and CF15-ΔF508CFTR (to test ER retention of ΔF508CFTR) cells in NF-κB activity, interleukin (IL)-8 secretion, Cai responses, and ER stress. Non-CF and CF primary cultures of human bronchial epithelial cells (HBE) secreted IL-8 equivalently. Upon infection with Pseudomonas aeruginosa (PA) or flagellin (key activator for airway epithelia), CF15, CF15-lacZ, CF15-wtCFTR, and CF15ΔF508CFTR cells exhibited equal PA binding, NF-κB activity, and IL-8 secretion; cells also responded similarly to flagellin when both CFTR (forskolin) and Cai signaling (ATP) were activated. CF and non-CF HBE responded similarly to flagellin + ATP. Thapsigargin (Tg, releases ER Ca2+) increased flagellin-stimulated NF-κB and ER stress similarly in all cells. We conclude that ER stress, Cai, and NF-κB signaling and IL-8 secretion were unaffected by wt- or ΔF508CFTR in control and during exposure to PA, flagellin, flagellin + ATP, or flagellin + ATP + forskolin. Tg, but not wt- or ΔF508CFTR, triggered ER stress. Previous measurements showing hyperinflammatory responses in CF airway epithelia may have resulted from cell-specific, rather than CFTR- or ΔF508CFTR-specific effects.

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