title: Smoke extract stimulates lung fibroblasts to release neutrophil and monocyte chemotactic activities.
abstract: Accumulation of monocytes and neutrophils and fibrous distortion of the airway are characteristics of airway disease secondary to smoking. The presence of inflammatory cells and fibrosis correlate, and, therefore, we postulated that lung fibroblasts might release chemotactic activity for neutrophils and monocytes in response to smoke extract. To test this hypothesis, human fetal lung (HFL1) fibroblasts were cultured, and the supernatant fluid was evaluated for neutrophil (NCA) and monocyte (MCA) chemotactic activities with a blind well chamber technique. HFL1 fibroblasts released chemotactic activity in response to smoke extract in a dose- and time-dependent manner (P < 0.05). Checkerboard analysis showed that the activity was predominantly chemotactic. Partial characterization of the released chemotactic activity revealed that the activity was partly heat labile, trypsin sensitive, and ethyl acetate extractable. Lipoxygenase inhibitors and cycloheximide inhibited the release of both NCA and MCA. Molecular-sieve chromatography revealed that NCA and MCA were heterogeneous. NCA was inhibited by anti-human interleukin (IL)-8 and anti-granulocyte colony-stimulating factor antibodies and a leukotriene (LT) B(4)-receptor antagonist. Anti-granulocyte-macrophage colony-stimulating factor (GM-CSF) and anti-monocyte chemoattractant protein (MCP)-1 antibodies and an LTB(4)-receptor antagonist inhibited MCA. Immunoreactive IL-8, granulocyte colony-stimulating factor, GM-CSF, and MCP-1 significantly increased in culture supernatant fluid in response to smoke extract. Finally, smoke extract augmented the expression of mRNAs of IL-8, GM-CSF, and MCP-1. These data demonstrate that lung fibroblasts release NCA and MCA in response to smoke extract and suggest that lung fibroblasts may modulate the inflammatory cell recruitment into the lung.
authors.affiliation: The First Department of Internal Medicine, Shinshu University School of Medicine, Matsumoto 390-8621, Japan.
authors.forename: E
authors.initials: E
authors.lastname: Sato
authors.forename: S
authors.initials: S
authors.lastname: Koyama
authors.forename: A
authors.initials: A
authors.lastname: Takamizawa
authors.forename: T
authors.initials: T
authors.lastname: Masubuchi
authors.forename: K
authors.initials: K
authors.lastname: Kubo
authors.forename: R A
authors.initials: RA
authors.lastname: Robbins
authors.forename: S
authors.initials: S
authors.lastname: Nagai
authors.forename: T
authors.initials: T
authors.lastname: Izumi
mesh_terms: D002478:Cells, Cultured; D018932:Chemokine CCL2 / Q000235:genetics; D018946:Chemokine CCL5 / Q000378:metabolism; D002633:Chemotaxis / Q000187:drug effects* / Q000276:immunology; D002845:Chromatography / Q000379:methods; D003513:Cycloheximide / Q000494:pharmacology; D005347:Fibroblasts / Q000166:cytology; D015870:Gene Expression / Q000187:drug effects / Q000276:immunology; D016179:Granulocyte Colony-Stimulating Factor / Q000235:genetics; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000235:genetics; D006801:Humans; D007166:Immunosuppressive Agents / Q000494:pharmacology; D016209:Interleukin-8 / Q000235:genetics; D007546:Isoquinolines / Q000494:pharmacology; D007975:Leukotriene B4 / Q000494:pharmacology; D008168:Lung / Q000276:immunology / Q000473:pathology*; D009000:Monocytes / Q000166:cytology / Q000276:immunology*; D009504:Neutrophils / Q000166:cytology / Q000276:immunology*; D010666:Phenylpropionates / Q000494:pharmacology; D010947:Plants, Toxic; D010975:Platelet Aggregation Inhibitors / Q000494:pharmacology; D010980:Platelet Membrane Glycoproteins / Q000037:antagonists & inhibitors; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D011658:Pulmonary Fibrosis / Q000139:chemically induced / Q000276:immunology / Q000473:pathology; D011726:Pyridinium Compounds / Q000494:pharmacology; D011956:Receptors, Cell Surface; D043562:Receptors, G-Protein-Coupled; D012907:Smoking / Q000009:adverse effects / Q000276:immunology*; D044005:Tetrahydroisoquinolines; D014026:Nicotiana / Q000009:adverse effects; D016212:Transforming Growth Factor beta / Q000378:metabolism
chemical_list: D018932:Chemokine CCL2; D018946:Chemokine CCL5; D007166:Immunosuppressive Agents; D016209:Interleukin-8; D007546:Isoquinolines; D010666:Phenylpropionates; D010975:Platelet Aggregation Inhibitors; D010980:Platelet Membrane Glycoproteins; D011500:Protein Synthesis Inhibitors; D011726:Pyridinium Compounds; D011956:Receptors, Cell Surface; D043562:Receptors, G-Protein-Coupled; D044005:Tetrahydroisoquinolines; D016212:Transforming Growth Factor beta; C039351:platelet activating factor receptor; C067523:ONO-LB 457; D016179:Granulocyte Colony-Stimulating Factor; D007975:Leukotriene B4; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D003513:Cycloheximide; C067430:TCV 309