title: Cigarette smoke and platelet-activating factor receptor dependent adhesion of Streptococcus pneumoniae to lower airway cells.
abstract: BACKGROUND
  Exposure to cigarette smoke (CS) is associated with increased risk of pneumococcal infection. The mechanism for this association is unknown. We recently reported that the particulate matter from urban air simulates platelet-activating factor receptor (PAFR)-dependent adhesion of pneumococci to airway cells. We therefore sought to determine whether CS stimulates pneumococcal adhesion to airway cells.
  
  
  METHODS
  Human alveolar (A549), bronchial (BEAS2-B), and primary bronchial epithelial cells (HBEpC) were exposed to CS extract (CSE), and adhesion of Streptococcus pneumoniae determined. The role of PAFR in mediating adhesion was determined using a blocker (CV-3988). PAFR transcript level was assessed by quantitative real-time PCR, and PAFR expression by flow cytometry. Lung PAFR transcript level was assessed in mice exposed to CS, and bronchial epithelial PAFR expression assessed in active-smokers by immunostaining.
  
  
  RESULTS
  In A549 cells, CSE 1% increased pneumococcal adhesion (p<0.05 vs control), PAFR transcript level (p<0.01), and PAFR expression (p<0.01). Pneumococcal adhesion to A549 cells was attenuated by CV-3988 (p<0.001). CSE 1% stimulated pneumococcal adhesion to BEAS2-B cells and HBEpC (p<0.01 vs control). CSE 1% increased PAFR expression in BEAS2-B (p<0.01), and in HBEpC (p<0.05). Lung PAFR transcript level was increased in mice exposed to CS in vivo (p<0.05 vs room air). Active smokers (n=16) had an increased percentage of bronchial epithelium with PAFR-positive cells (p<0.05 vs never smokers, n=11).
  
  
  CONCLUSIONS
  CSE stimulates PAFR-dependent pneumococcal adhesion to lower airway epithelial cells. We found evidence that CS increases bronchial PAFR in vivo.
authors.affiliation: Centre for Paediatrics, Blizard Institute, Barts and the London School of Medicine and Dentistry, Queen Mary, University of London, 4 Newark Street, London E1 2AT, UK. j.grigg@qmul.ac.uk
authors.forename: Jonathan
authors.initials: J
authors.lastname: Grigg
authors.forename: Haydn
authors.initials: H
authors.lastname: Walters
authors.forename: Sukhwinder Singh
authors.initials: SS
authors.lastname: Sohal
authors.forename: Richard
authors.initials: R
authors.lastname: Wood-Baker
authors.forename: David W
authors.initials: DW
authors.lastname: Reid
authors.forename: Cang-Bao
authors.initials: CB
authors.lastname: Xu
authors.forename: Lars
authors.initials: L
authors.lastname: Edvinsson
authors.forename: Mathieu C
authors.initials: MC
authors.lastname: Morissette
authors.forename: Martin R
authors.initials: MR
authors.lastname: Stämpfli
authors.forename: Michael
authors.initials: M
authors.lastname: Kirwan
authors.forename: Lee
authors.initials: L
authors.lastname: Koh
authors.forename: Reetika
authors.initials: R
authors.lastname: Suri
authors.forename: Naseem
authors.initials: N
authors.lastname: Mushtaq
mesh_terms: D000818:Animals; D001422:Bacterial Adhesion / Q000187:drug effects* / Q000502:physiology*; D002460:Cell Line; D005434:Flow Cytometry; D006801:Humans; D066298:In Vitro Techniques; D051379:Mice; D052638:Particulate Matter / Q000378:metabolism / Q000494:pharmacology; D010980:Platelet Membrane Glycoproteins / Q000378:metabolism*; D011008:Pneumococcal Infections / Q000209:etiology*; D060888:Real-Time Polymerase Chain Reaction; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism*; D012137:Respiratory System / Q000166:cytology* / Q000187:drug effects / Q000382:microbiology; D012907:Smoking / Q000378:metabolism*; D018709:Statistics, Nonparametric; D013296:Streptococcus pneumoniae / Q000378:metabolism; D014028:Tobacco Smoke Pollution / Q000009:adverse effects*
chemical_list: D052638:Particulate Matter; D010980:Platelet Membrane Glycoproteins; D043562:Receptors, G-Protein-Coupled; D014028:Tobacco Smoke Pollution; C039351:platelet activating factor receptor