Organic dust activates NF-kappaB in lung epithelial cells.
Exposure in swine confinement facilities induces airway inflammation in healthy subjects. The aim of the present study was to elucidate the role of nuclear factor (NF)-kappaB in the inflammatory response induced by organic dust. A human lung epithelial carcinoma cell line (A549) was transfected with reporter genes of the human IL-6 promoter or the NF-kappaB binding site fused to the luciferase reporter gene and stimulated with dust from a swine confinement building. Cytokine release in cell culture supernatants and luciferase activity was measured. The dust-induced the activities of the IL-6 promoter reporter gene and the NF-kappaB reporter gene in parallel with an increase in IL-6 and IL-8 release. The addition of pyrrolidinedithiocarbamate, a chemical NF-kappaB blocking agent, inhibited IL-6 and IL-8 secretion as well as the NF-kappaB reporter gene activity. Increasing the amount of IkappaB alpha led to inhibition of organic dust-induced IL-6 promoter and NF-kappaB reporter gene activities. Fluticasone inhibited the organic dust-induced NF-kappaB activation and IL-6 and IL-8 secretion. Finally, swine dust incubation of A549 cells resulted in a NF-kappaB DNA binding, which is composed of the NF-kappaB1 and RelA proteins. In conclusion, by interference at various levels we have shown that NF-kappaB plays a key role in the inflammatory response to organic dust.
Lidén
Johan
J
Department of Medical Nutrition, Karolinska Institutet, Huddinge University Hospital Novum, Huddinge SE-141 86, Sweden.
Ek
Alexandra
A
Palmberg
Lena
L
Okret
Sam
S
Larsson
Kjell
K
D000730:Androstadienes / Q000494:pharmacology; D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology; D004391:Dust; D004586:Electrophoresis; D004847:Epithelial Cells / Q000378:metabolism*; D000068298:Fluticasone; D017930:Genes, Reporter / Q000502:physiology; D006801:Humans; D015850:Interleukin-6 / Q000378:metabolism; D016209:Interleukin-8 / Q000378:metabolism; D008156:Luciferases / Q000378:metabolism*; D008168:Lung / Q000166:cytology / Q000378:metabolism*; D016328:NF-kappa B / Q000378:metabolism*; D013388:Succinimides / Q000494:pharmacology; D013552:Swine; D014162:Transfection; D014407:Tumor Cells, Cultured
D000730:Androstadienes; D000893:Anti-Inflammatory Agents; D004391:Dust; D015850:Interleukin-6; D016209:Interleukin-8; D016328:NF-kappa B; D013388:Succinimides; D000068298:Fluticasone; D008156:Luciferases