Effects of 0.2 ppm ozone on biomarkers of inflammation in bronchoalveolar lavage fluid and bronchial mucosa of healthy subjects.
Short-term exposure to ozone at peak ambient levels induces neutrophil influx and impairs lung function in healthy humans. In order to investigate the mechanisms contributing to neutrophil recruitment and to examine the role of T-cells in the acute inflammatory response, we exposed 12 healthy humans to 0.2 parts per million (ppm) of ozone and filtered air on two separate occasions for 2 h with intermittent periods of rest and exercise (minute ventilation = 30 L x min(-1)). Fibreoptic bronchoscopy was performed 6 h after the end of exposures. Total protein, tryptase, histamine, myeloperoxidase, interleukin (IL)-8 and growth-related oncogene-alpha (Gro-alpha) were measured and total and differential cell counts were performed in bronchoalveolar lavage (BAL) fluid. Flow cytometry was performed on BAL cells to study total T-cells, T-cell receptors (alphabeta and gammadelta), T-cell subsets (CD4+ and CD8+ cells) and activated T-cell subsets (CD25+). Using immunohistochemistry, neutrophils, mast cells, total T-cell numbers, T-cell subsets, CD25+ T-cells and leukocyte endothelial adhesion molecules including P-selectin, E-selectin, intercellular adhesion molecule (ICAM)-1 and vascular adhesion molecule (VCAM)-1 were quantified in the bronchial biopsies. Paired samples were available from nine subjects. Following ozone exposure there was a threefold increase in the proportion of polymorphonuclear neutrophils (PMNs) (p=0.07) and epithelial cells (p=0.05) in BAL fluid. This was accompanied by increased concentrations of IL-8 (p=0.01), Gro-alpha (p=0.05) and total protein (p=0.058). A significant positive correlation was demonstrated between the two chemokines and proportion of PMNs in BAL fluid. After ozone exposure there was a significant decrease in the CD4/CD8 ratio (p=0.05) and the proportion of activated CD4+ (p=0.01) and CD8+ T-cells (p=0.04). However, no significant changes were demonstrable in any of the inflammatory markers studied in the biopsies. Short-term exposure of healthy humans to 0.2 ppm ozone induced a neutrophil influx in peripheral airways at 6 h post exposure, but no apparent inflammatory response in proximal airways. This response seems to be mediated at least in part by interleukin-8 and growth-related oncogene-alpha.
Krishna
M T
MT
University Medicine, Southampton General Hospital, University of Southampton, UK.
Madden
J
J
Teran
L M
LM
Biscione
G L
GL
Lau
L C
LC
Withers
N J
NJ
Sandström
T
T
Mudway
I
I
Kelly
F J
FJ
Walls
A
A
Frew
A J
AJ
Holgate
S T
ST
D000328:Adult; D001980:Bronchi / Q000187:drug effects / Q000473:pathology*; D001992:Bronchoalveolar Lavage Fluid / Q000737:chemistry* / Q000166:cytology*; D015815:Cell Adhesion Molecules / Q000032:analysis; D002452:Cell Count; D053818:Chymases; D018592:Cross-Over Studies; D004311:Double-Blind Method; D005260:Female; D005434:Flow Cytometry; D006632:Histamine / Q000032:analysis; D006801:Humans; D007150:Immunohistochemistry; D018836:Inflammation Mediators / Q000032:analysis*; D016209:Interleukin-8 / Q000032:analysis; D008297:Male; D009092:Mucous Membrane / Q000187:drug effects / Q000473:pathology; D009504:Neutrophils; D015513:Oncogene Proteins / Q000032:analysis; D010083:Oxidants, Photochemical / Q000494:pharmacology*; D010126:Ozone / Q000494:pharmacology*; D009195:Peroxidase / Q000032:analysis; D015375:Receptors, Interleukin-2 / Q000032:analysis; D012697:Serine Endopeptidases / Q000032:analysis; D016176:T-Lymphocyte Subsets; D053802:Tryptases
D015815:Cell Adhesion Molecules; D018836:Inflammation Mediators; D016209:Interleukin-8; D015513:Oncogene Proteins; D010083:Oxidants, Photochemical; D015375:Receptors, Interleukin-2; D010126:Ozone; D006632:Histamine; D009195:Peroxidase; D012697:Serine Endopeptidases; C083294:chymase 2; D053818:Chymases; D053802:Tryptases