GM-CSF induction in human lung fibroblasts by IL-1beta, TNF-alpha, and macrophage contact.
Fibroblast-derived cytokines may play crucial roles in airway inflammation. In this study, we analyzed expression of the inflammatory cytokine, granulocyte-macrophage colony-stimulating factor (GM-CSF), a major eosinophilopoietin, by normal human lung fibroblast (NHLF) cells and its regulation by monokines and macrophage contact. NHLFs were stimulated with interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha) and were cocultured with the U937 myelomonocytic cell line. The expression of GM-CSF transcripts was analyzed by reverse transcription-polymerase chain reaction (RT-PCR), and GM-CSF protein was detected by ELISA. Nuclear translocation of nuclear factor-kappaB (NF-kappaB), an important transcription factor for inflammatory gene expression, was assessed by electrophoretic mobility shift assay (EMSA). Both IL-1beta and TNF-alpha significantly enhanced the production of GM-CSF by NHLF. Coculturing of peripheral blood mononuclear cells (PBMC) with NHLF induced GM-CSF expression. This phenomenon was also seen on coculturing U937 cells or membranes derived from U937 with NHLF but was inhibited when the two types of cells were separated, suggesting a need for cell-cell contact. U937 membranes, as well as IL-1beta and TNF-alpha, induced nuclear translocation of NF-kappaB. These data support a prominent role for macrophage-fibroblast interactions in airway inflammation and fibrosis.
Department of Internal Medicine, Division of Allergy and Immunology, East Tennessee State University, Quillen College of Medicine, Johnson City, TN 37614, USA.
S Matthew
SM
Fitzgerald
David S
DS
Chi
H Kenton
HK
Hall
Scott A
SA
Reynolds
Omolola
O
Aramide
Steven A
SA
Lee
Guha
G
Krishnaswamy
D000911:Antibodies, Monoclonal / Q000494:pharmacology; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism; D018920:Coculture Techniques; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects*; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007375:Interleukin-1 / Q000494:pharmacology*; D007963:Leukocytes, Mononuclear / Q000378:metabolism; D008168:Lung / Q000166:cytology / Q000378:metabolism*; D008264:Macrophages / Q000378:metabolism*; D008954:Models, Biological; D016328:NF-kappa B / Q000187:drug effects / Q000378:metabolism; D015398:Signal Transduction; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*; D020298:U937 Cells
D000911:Antibodies, Monoclonal; D007375:Interleukin-1; D016328:NF-kappa B; D014409:Tumor Necrosis Factor-alpha; D016178:Granulocyte-Macrophage Colony-Stimulating Factor