Neutrophil chemokines in cultured nasal fibroblasts

• Published in: Allergy (Copenhagen) Vol. 57; no. 12; pp. 1159 - 1164
• Main Authors: Rudack, C., Hermann, W., Eble, J., Schroeder, J.‐M.
• Format: Journal Article
• Language: English
• Published: Oxford, UK Munksgaard International Publishers 01.12.2002; Blackwell
• Subjects: Allergic diseases; Antimicrobial Cationic Peptides; biological activity; Biological and medical sciences; Blood Proteins - pharmacology; Carrier Proteins - pharmacology; Cell Movement - drug effects; Cell Movement - immunology; Cells, Cultured; Chemokine CXCL1; chemokines; Chemokines - biosynthesis; Chemokines - immunology; Chemokines, CXC - immunology; Chemokines, CXC - metabolism; Chemotactic Factors - biosynthesis; Chemotactic Factors - immunology; Chromatography; Dose-Response Relationship, Drug; Enzyme-Linked Immunosorbent Assay; Fibroblasts - immunology; Fibroblasts - metabolism; Humans; Immunopathology; Intercellular Signaling Peptides and Proteins - biosynthesis; Intercellular Signaling Peptides and Proteins - immunology; Interleukin-8 - biosynthesis; Interleukin-8 - immunology; Interleukin‐8; Medical sciences; nasal fibroblast; Nasal Mucosa - cytology; Nasal Mucosa - immunology; Nasal Mucosa - metabolism; Neutrophils - immunology; Neutrophils - metabolism; Polymerase Chain Reaction; Respiratory and ent allergic diseases; RNA, Messenger - drug effects; RNA, Messenger - immunology; RNA, Messenger - metabolism; Stimulation, Chemical; Time Factors; Tumor Necrosis Factor-alpha - administration & dosage; Up-Regulation - drug effects; Up-Regulation - physiology; Human; Cell culture; Pathogenesis; Cytokine; chemokines; Interleukin-8; Biological activity; Sinusitis; Chemokine; ENT disease; Paranasal sinus disease; Molecular biology; Nasal fossa; Fibroblast; Reverse transcription polymerase chain reaction; ELISA assay; Interleukin 8; nasal fibroblast
• ISSN: 0105-4538; 1398-9995
• DOI: 10.1034/j.1398-9995.2002.23748.x

Background: To gain insight into the mechanisms responsible for tissue neutrophil immigration in sinusitis, primary nasal fibroblasts are analyzed for synthesizing and delivering neutrophil chemokines. Methods: Primary nasal fibroblast cell culture was treated with tumor necrosis factor (TNF)‐α concentrations of 20 and 200 ng/ml for 2, 8, 24 and 72 h. Chemokine concentrations in supernatants were determined by enzyme‐linked immunoassay (ELISA) and chemokine mRNA expression in fibroblasts was measured by reverse transcriptase polymerase chain reaction (RT‐PCR). Biological chemotactic activity was identified by three‐step high‐performance liquid chromatography (HPLC) and by bioassay measuring neutrophil chemotaxis in a single Boyden chamber system. Results: Interleukin (IL)‐8 and growth‐related oncogene (GRO)‐α were induced in nasal fibroblast culture by proinflammatory stimulus. After 24 h of stimulation neutrophil chemotactic activity only was detected for IL‐8. Granulocyte chemotactic protein (GCP)‐2 mRNA was already significantly up‐regulated after 2 h of stimulation. Conclusion: Induction of IL‐8 protein dominates chemokine synthesis 24 and 72 h after stimulation, whereas induction of GCP‐2 mRNA seems to have a role in the early phase after 2 h of exposition with TNF‐α.