title: Toll-like receptor 4 is not targeted to the lysosome in cystic fibrosis airway epithelial cells.
abstract: The innate immune response to bacterial infection is mediated through Toll-like receptors (TLRs), which trigger tightly regulated signaling cascades through transcription factors including NF-κB. LPS activation of TLR4 triggers internalization of the receptor-ligand complex which is directed toward lysosomal degradation or endocytic recycling. Cystic fibrosis (CF) patients display a robust and uncontrolled inflammatory response to bacterial infection, suggesting a defect in regulation. This study examined the intracellular trafficking of TLR4 in CF and non-CF airway epithelial cells following stimulation with LPS. We employed cells lines [16hBE14o-, CFBE41o- (CF), and CFTR-complemented CFBE41o-] and confirmed selected experiments in primary nasal epithelial cells from non-CF controls and CF patients (F508del homozygous). In control cells, TLR4 expression (surface and cytoplasmic) was reduced after LPS stimulation but remained unchanged in CF cells and was accompanied by a heightened inflammatory response 24 h after stimulation. All cells expressed markers of the early (EEA1) and late (Rab7b) endosomes at basal levels. However, only CF cells displayed persistent expression of Rab7b following LPS stimulation. Rab7 variants may directly internalize bacteria to the Golgi for recycling or to the lysosome for degradation. TLR4 colocalized with the lysosomal marker LAMP1 in 16 hBE14o- cells, suggesting that TLR4 is targeted for lysosomal degradation in these cells. However, this colocalization was not observed in CFBE41o- cells, where persistent expression of Rab7 and release of proinflammatory cytokines was detected. Consistent with the apparent inability of CF cells to target TLR4 toward the lysosome for degradation, we observed persistent surface and cytoplasmic expression of this pathogen recognition receptor. This defect may account for the prolonged cycle of chronic inflammation associated with CF.
authors: Centre for Infection and Immunity, Queen's University of Belfast, Belfast, UK.
authors: Catriona
authors: C
authors: Kelly
authors: Paul
authors: P
authors: Canning
authors: Paul J
authors: PJ
authors: Buchanan
authors: Mark T
authors: MT
authors: Williams
authors: Vanessa
authors: V
authors: Brown
authors: Dieter C
authors: DC
authors: Gruenert
authors: J Stuart
authors: JS
authors: Elborn
authors: Madeleine
authors: M
authors: Ennis
authors: Bettina C
authors: BC
authors: Schock
mesh_terms: D001980:Bronchi / Q000166:cytology / Q000276:immunology*; D002460:Cell Line; D003550:Cystic Fibrosis / Q000276:immunology* / Q000378:metabolism* / Q000473:pathology; D011992:Endosomes / Q000378:metabolism; D004847:Epithelial Cells / Q000378:metabolism*; D006801:Humans; D007249:Inflammation / Q000276:immunology; D008070:Lipopolysaccharides / Q000276:immunology; D051907:Lysosomal Membrane Proteins / Q000378:metabolism; D008247:Lysosomes / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism; D021381:Protein Transport; D011552:Pseudomonas Infections / Q000276:immunology; D011550:Pseudomonas aeruginosa / Q000276:immunology*; D020545:Respiratory Mucosa / Q000166:cytology / Q000276:immunology*; D051197:Toll-Like Receptor 4 / Q000378:metabolism*; D033921:Vesicular Transport Proteins / Q000096:biosynthesis; D020691:rab GTP-Binding Proteins / Q000096:biosynthesis; D000090802:rab7 GTP-Binding Proteins; D052118:Lysosomal-Associated Membrane Protein 1
chemical_list: D008070:Lipopolysaccharides; D051907:Lysosomal Membrane Proteins; D008565:Membrane Proteins; D051197:Toll-Like Receptor 4; D033921:Vesicular Transport Proteins; D020691:rab GTP-Binding Proteins; D000090802:rab7 GTP-Binding Proteins; C489116:LAMP1 protein, human; D052118:Lysosomal-Associated Membrane Protein 1; C495345:TLR4 protein, human; C093846:early endosome antigen 1; C000716463:rab7 GTP-binding proteins, human