Neutrophil chemokines in cultured nasal fibroblasts
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 conc...
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Published in | Allergy (Copenhagen) Vol. 57; no. 12; pp. 1159 - 1164 |
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Main Authors | , , , |
Format | Journal Article |
Language | English |
Published |
Oxford, UK
Munksgaard International Publishers
01.12.2002
Blackwell |
Subjects | |
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Abstract | 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. |
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AbstractList | 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. 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. To gain insight into the mechanisms responsible for tissue neutrophil immigration in sinusitis, primary nasal fibroblasts are analyzed for synthesizing and delivering neutrophil chemokines. Primary nasal fibroblast cell culture was treated with tumor necrosis factor (TNF)-alpha 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. Interleukin (IL)-8 and growth-related oncogene (GRO)-alpha 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. 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-alpha. |
Author | Rudack, C. Schroeder, J.‐M. Eble, J. Hermann, W. |
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Keywords | 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 |
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Snippet | Background: To gain insight into the mechanisms responsible for tissue neutrophil immigration in sinusitis, primary nasal fibroblasts are analyzed for... To gain insight into the mechanisms responsible for tissue neutrophil immigration in sinusitis, primary nasal fibroblasts are analyzed for synthesizing and... Background: To gain insight into the mechanisms responsible for tissue neutrophil immigration in sinusitis, primary nasal fibroblasts are analyzed for... |
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SubjectTerms | 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 |
Title | Neutrophil chemokines in cultured nasal fibroblasts |
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