title: TLR expression on neutrophils at the pulmonary site of infection: TLR1/TLR2-mediated up-regulation of TLR5 expression in cystic fibrosis lung disease.
abstract: Cystic fibrosis (CF) lung disease is characterized by infection with Pseudomonas aeruginosa and a sustained accumulation of neutrophils. In this study, we analyzed 1) the expression of MyD88-dependent TLRs on circulating and airway neutrophils in P. aeruginosa-infected CF patients, P. aeruginosa-infected non-CF bronchiectasis patients, and noninfected healthy control subjects and 2) studied the regulation of TLR expression and functionality on neutrophils in vitro. TLR2, TLR4, TLR5, and TLR9 expression was increased on airway neutrophils compared with circulating neutrophils in CF and bronchiectasis patients. On airway neutrophils, TLR5 was the only TLR that was significantly higher expressed in CF patients compared with bronchiectasis patients and healthy controls. Studies using confocal microscopy and flow cytometry revealed that TLR5 was stored intracellularly in neutrophils and was mobilized to the cell surface in a protein synthesis-independent manner through protein kinase C activation or after stimulation with TLR ligands and cytokines characteristic of the CF airway microenvironment. The most potent stimulator of TLR5 expression was the bacterial lipoprotein Pam(3)CSK(4). Ab-blocking experiments revealed that the effect of Pam(3)CSK(4) was mediated through cooperation of TLR1 and TLR2 signaling. TLR5 activation enhanced the phagocytic capacity and the respiratory burst activity of neutrophils, which was mediated, at least partially, via a stimulation of IL-8 production and CXCR1 signaling. This study demonstrates a novel mechanism of TLR regulation in neutrophils and suggests a critical role for TLR5 in neutrophil-P. aeruginosa interactions in CF lung disease.
authors.affiliation: Children's Hospital of the Ludwig-Maximilians-University, Munich, Germany.
authors.forename: Barbara
authors.initials: B
authors.lastname: Koller
authors.forename: Matthias
authors.initials: M
authors.lastname: Kappler
authors.forename: Phillip
authors.initials: P
authors.lastname: Latzin
authors.forename: Amit
authors.initials: A
authors.lastname: Gaggar
authors.forename: Marcus
authors.initials: M
authors.lastname: Schreiner
authors.forename: Sherkin
authors.initials: S
authors.lastname: Takyar
authors.forename: Michael
authors.initials: M
authors.lastname: Kormann
authors.forename: Michael
authors.initials: M
authors.lastname: Kabesch
authors.forename: Dirk
authors.initials: D
authors.lastname: Roos
authors.forename: Matthias
authors.initials: M
authors.lastname: Griese
authors.forename: Dominik
authors.initials: D
authors.lastname: Hartl
mesh_terms: D000328:Adult; D003550:Cystic Fibrosis / Q000276:immunology* / Q000382:microbiology / Q000473:pathology; D005260:Female; D006801:Humans; D007424:Intracellular Fluid / Q000276:immunology / Q000378:metabolism; D008168:Lung / Q000276:immunology* / Q000378:metabolism / Q000382:microbiology / Q000473:pathology; D008297:Male; D008565:Membrane Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D008875:Middle Aged; D009504:Neutrophils / Q000276:immunology* / Q000378:metabolism* / Q000382:microbiology / Q000473:pathology; D021381:Protein Transport / Q000276:immunology; D011550:Pseudomonas aeruginosa / Q000276:immunology; D020545:Respiratory Mucosa / Q000276:immunology / Q000378:metabolism; D051194:Toll-Like Receptor 1 / Q000502:physiology*; D051195:Toll-Like Receptor 2 / Q000502:physiology*; D051198:Toll-Like Receptor 5 / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D015854:Up-Regulation / Q000276:immunology*
chemical_list: D008565:Membrane Proteins; C495342:TLR2 protein, human; C495346:TLR5 protein, human; D051194:Toll-Like Receptor 1; D051195:Toll-Like Receptor 2; D051198:Toll-Like Receptor 5