title: Involvement of microbial components and toll-like receptors 2 and 4 in cytokine responses to air pollution particles.
abstract: Inhalation of particulate matter (PM) may result in exacerbation of inflammatory airways disease, including asthma. Results from this laboratory have shown that the coarse inhalable particle fraction (PM(2.5-10)) is responsible for most of the PM effects on human airway macrophages (AM), including induction of cytokine production. Endotoxins associated with these particles account for a large part of their potency, as activity of PM can be inhibited by polymixin B and an activating moiety bound by lipopolysaccharide (LPS)-binding protein (LBP). The hypothesis behind the present study was that not only particle-bound LPS, but also Gram-negative (Gram-) and Gram-positive (Gram+) bacteria are responsible for PM-induced stimulation of AM, and therefore that PM are likely to activate receptors involved in recognition of microbes. Low level contamination of model pollution particles with environmental Staphyloccocus, Streptococcus, and Pseudomonas species was found to confer cytokine-inducing activity on inactive particles. Only one Gram- bacterium was sufficient for significant stimulatation of 100 AM, whereas at least three times more Gram+ bacteria were required for a similar level of response. Cytokine responses induced by PM as well as Gram+ and Gram- bacteria were inhibited by anti-CD14 antibody and required the presence of LBP-containing serum. The involvement of Toll-like receptor (TLR) 2 and 4 in recognition of PM(2.5-10) was investigated in transfected Chinese hamster ovary cells expressing CD14 and TLR2 or TLR4. TLR4 was found to be involved in PM(2.5-10) and Pseudomonas-induced activation, whereas TLR2 activation was induced by both Gram+ and Gram- bacteria and by PM. The synthetic lipid A analog E5531 fully inhibited the response to purified LPS and partially inhibited the response to PM and Pseudomonas. In contrast, E5531 had no effect on the response to Staphylococcus. Taken together, these results implicate microbial components as important players in AM-dependent inflammatory responses to PM.
authors.affiliation: U.S. Environmental Protection Agency, National Health and Environmental Effects Research Laboratory, Research Triangle Park, North Carolina, USA. Becker.susanne@epa.gov
authors.forename: Susanne
authors.initials: S
authors.lastname: Becker
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Fenton
authors.forename: Joleen M
authors.initials: JM
authors.lastname: Soukup
mesh_terms: D000328:Adult; D000391:Air Microbiology; D000393:Air Pollutants / Q000737:chemistry / Q000633:toxicity*; D000818:Animals; D001419:Bacteria / Q000472:pathogenicity*; D016466:CHO Cells; D002478:Cells, Cultured; D006224:Cricetinae; D016207:Cytokines / Q000378:metabolism*; D029721:Drosophila Proteins; D005260:Female; D006801:Humans; D019570:Inhalation Exposure; D008050:Lipid A / Q000031:analogs & derivatives* / Q000494:pharmacology; D018950:Lipopolysaccharide Receptors / Q000187:drug effects / Q000378:metabolism; D008070:Lipopolysaccharides; D016676:Macrophages, Alveolar / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D008562:Membrane Glycoproteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011112:Polymyxin B / Q000494:pharmacology; D011549:Pseudomonas / Q000472:pathogenicity; D011956:Receptors, Cell Surface / Q000037:antagonists & inhibitors / Q000378:metabolism*; D013210:Staphylococcus / Q000472:pathogenicity; D013291:Streptococcus / Q000472:pathogenicity; D051195:Toll-Like Receptor 2; D051197:Toll-Like Receptor 4; D051193:Toll-Like Receptors
chemical_list: D000393:Air Pollutants; D016207:Cytokines; D029721:Drosophila Proteins; D008050:Lipid A; D018950:Lipopolysaccharide Receptors; D008070:Lipopolysaccharides; D008562:Membrane Glycoproteins; D011956:Receptors, Cell Surface; C495342:TLR2 protein, human; C495345:TLR4 protein, human; D051195:Toll-Like Receptor 2; D051197:Toll-Like Receptor 4; D051193:Toll-Like Receptors; C092845:E 5531; D011112:Polymyxin B