title: Effects of PM10 in human peripheral blood monocytes and J774 macrophages.
abstract: The effects of PM10, one of the components of particulate air pollution, was investigated using human monocytes and a mouse macrophage cell line (J774). The study aimed to investigate the role of these nanoparticles on the release of the pro-inflammatory cytokine TNF-alpha and IL-1alpha gene expression. We also investigated the role of intracellular calcium signalling events and oxidative stress in control of these cytokines and the effect of the particles on the functioning of the cell cytoskeleton. We showed that there was an increase in intracellular calcium concentration in J774 cells on treatment with PM10 particles which could be significantly reduced with concomitant treatment with the calcium antagonists verapamil, the intracellular calcium chelator BAPTA-AM but not with the antioxidant nacystelyn or the calmodulin inhibitor W-7. In human monocytes, PM10 stimulated an increase in intracellular calcium which was reduced by verapamil, BAPTA-AM and nacystelyn. TNF-alpha release was increased with particle treatment in human monocytes and reduced by only verapamil and BAPTA-AM. IL-1alpha gene expression was increased with particle treatment and reduced by all of the inhibitors. There was increased F-actin staining in J774 cells after treatment with PM10 particles, which was significantly reduced to control levels with all the antagonists tested. The present study has shown that PM10 particles may exert their pro-inflammatory effects by modulating intracellular calcium signalling in macrophages leading to expression of pro-inflammatory cytokines. Impaired motility and phagocytic ability as shown by changes in the F-actin cytoskeleton is likely to play a key role in particle clearance from the lung.
authors.affiliation: School of Life Sciences, Napier University, Edinburgh, UK. da.brown@napier.ac.uk
authors.forename: D M
authors.initials: DM
authors.lastname: Brown
authors.forename: K
authors.initials: K
authors.lastname: Donaldson
authors.forename: V
authors.initials: V
authors.lastname: Stone
mesh_terms: D000393:Air Pollutants / Q000737:chemistry / Q000633:toxicity*; D000818:Animals; D020013:Calcium Signaling / Q000187:drug effects / Q000276:immunology*; D002460:Cell Line; D016207:Cytokines / Q000276:immunology; D003598:Cytoskeletal Proteins / Q000276:immunology; D003599:Cytoskeleton / Q000187:drug effects / Q000276:immunology; D005786:Gene Expression Regulation / Q000187:drug effects / Q000276:immunology; D006801:Humans; D007375:Interleukin-1 / Q000276:immunology*; D007963:Leukocytes, Mononuclear / Q000187:drug effects / Q000276:immunology*; D008262:Macrophage Activation / Q000187:drug effects / Q000276:immunology; D008264:Macrophages / Q000187:drug effects / Q000276:immunology* / Q000378:metabolism; D051379:Mice; D049329:Nanostructures / Q000737:chemistry / Q000633:toxicity*; D010316:Particle Size; D014409:Tumor Necrosis Factor-alpha / Q000276:immunology*; D001335:Vehicle Emissions / Q000633:toxicity
chemical_list: D000393:Air Pollutants; D016207:Cytokines; D003598:Cytoskeletal Proteins; D007375:Interleukin-1; D014409:Tumor Necrosis Factor-alpha; D001335:Vehicle Emissions